Aegean Conferences Series-Vol. 54




Linking the international scientific community

Bringing the humanity scholars together


together with the






24th International Mammalian

Genome Conference













October 17-21, 2010

Aldemar: Knossos Royal Village Conference Center

Hersonissos, Crete, Greece

24th International Mammalian Genome Conference

Aldemar Knossos Royal Village

October 17-21, 2010

Hersonissos, Crete, Greece




International Mammalian Genome Society Secretariat


Maja Bucan, Past President (2010)

Karen Steel, President (2010)

David Threadgill, Vice-President (President 2011)


Kent Hunter (2010)

David Beier(2011)

Ian Jackson(2011)

Nancy Jenkins (2011)

Simon Foote(2012)

Xavier Montagutelli (2012)

Darren Logan (2011) VM Chapman award winner



Nomination and Election Committee


Kathryn Hentges (2010)

Bruce Herron (2010)

Amy Moser (2010)

Rosemary Elliott (2011)

Beverly Mock (2011)

Thomas Saunders (2011)


Committee on the Standardized Genetic Nomenclature for Mice


Janan Eppig, Chair


Ruth Arkell                    Simon Foote                  Amy Moser

Piero Carninci                 Jiri Forejt                       Jean-Jacques Panthier

Sally Cross                    Teresa Gunn                   David Threadgill

Deanna Church                David Largespada            Wolfgang Wurst

Jonathan Flint                 Lois Maltais                   Aamir Zuberi


Official Journal of the

International Mammalian Genome Society


Sponsored by the

International Committee for

Standardized Genetic Nomenclature


Joseph H. Nadeau

Genetics Department

Case Western Reserve University

10900 Euclid Avenue

Cleveland, Ohio 44106-4955



Steve D.M. Brown

Medical Research Council

Mammalian Genetics Unit

Harwell, Didcot

Oxfordshire OX11 ORD

United Kingdom


Informatics Editors

Janan T. Eppig, Bar Harbor, Maine USA



Reviews Editor

Maja Bucan, Philadelphia, Pennsylvania, USA


Managing Editor                      

Louise Tinsley, Oxfordshire, UK



IMGS Business Manager

Darla Miller, Chapel Hill, NC, USA


D. Adams, Cambridge, United Kingdom

M.D. Adams, Cleveland, Ohio, USA

T. Aitman, London, United Kingdom

L. Andersson, Uppsala, Sweden

R. Arkell, Canberra, Australia

P. Avner, Paris, France

E. Bailey, Lexington, Kentucky, USA

R. Balling, Braunschweig, Germany

D.R. Beier, Boston, Massachusetts, USA

B. Beutler, La Jolla, California, USA

S. Camper, Ann Arbor, Michigan, USA

G. Churchill, Bar Harbor, Maine, USA

N.G. Copeland, Singapore

E. Fisher, London, United Kingdom

S. Foote, Hobart, Tasmania, Australia

W. Frankel, Bar Harbor, Maine, USA

D. Gauguier, Oxford, United Kingdom

M. Georges, Liege, Belgium

C. Goodnow, Canberra, Australia

K.A. Gould, Omaha, Nebraska, USA

E.D. Green, Bethesda, Maryland, USA

P. Gros, Montreal, Quebec, Canada

Y. Herault, Orleans, France


M. Hrabe« de Angelis, Neuherberg, Germany

D.A. Hume, Edinburgh, UK

I.J. Jackson, Edinburgh, United Kingdom

N. Jenkins, Singapore

M. Justice, Houston, Texas, USA

G. Kelsey, Cambridge, UK

H. Khatib, Madison, Wisconsin, USA

E. Lander, Cambridge, Massachusetts, USA

M. Lyon, Oxfordshire, United Kingdom

T. Magnuson, Chapel Hill, NC, USA

K. Murphy, College Station, Texas, USA

D. Norris, Oxfordshire, United Kingdom

E.A. Ostrander, Bethesda, Maryland, USA

W.J. Pavan, Bethesda, Maryland, USA

D. Pomp, Chapel Hill, North Carolina, USA

L.C. Schalkwyk, London, United Kingdom

T. Shiroishi, Shizuoka-ken, Japan

K. Steel, Cambridge, United Kingdom

C.K. Tuggle, Ames, Iowa, USA

E. Whitelaw, Brisbane, Australia

L. Wicker, Cambridge, United Kingdom

J. Womack, College Station, Texas, USA

J.G. Zhang, Ann Arbor, Michigan, USA




Program Outline






Bio Informatics Workshop Program


Conference Map


Hotel Map




List of Participants


Upcoming Events






The Secretariat and members of The International Mammalian Genome Society are proud of the students, postdoctoral fellows and junior faculty who present oral and poster presentations at the annual meeting.   The outstanding contributions of these young scientists are recognized through several awards.


The first of these awards is the Verne Chapman Young Scientist Award, which is given to the most outstanding oral presentation from a postdoctoral fellow or student.  This is a monetary award of $500 that reflects Dr. ChapmanÕs dedication to mentoring junior scientists.  This award also includes a two year term on the secretariat.  Two second place monetary awards of $250 each are given this year by The Genetics Society of America.


A large group of publishing companies sponsor this yearÕs presentation prizes, which are a one year subscription to their journal. Genesis is donating books.  These awards are given to the most outstanding poster presentations by graduate students and postdoctoral fellows, and are chosen by members of the IMGS Secretariat during the course of the meeting.


This year the sponsors are:

Description: genomics_logo


Description: GSAlogo





Genetics Research from

Description: Macintosh Journals logo.TIF



Funding for student scholarships was made possible by:
2R13HG002394 from the following ICs at NIH:  NHGRI, NIMH, NICHD, NIAID, NIEHS, NCRR, AND NINDS





The conference participants will be staying at the Aldemar: Knossos Royal Village (A' class), which is located in Hersonissos, 25 km from the airport.


Arrival and Check-in

It is expected that attendees will arrive/check-in on Sunday after 2:00 PM. Anyone arriving after 10:30 pm on Sunday must make alternative dinner arrangements.


Conference Venue

The Aldemar: Knossos Royal Village Conference Center is located at the hotel.  (#29 on Hotel Map)


Handicapped Accessibility

The hotel and conference facilities are handicapped accessible.



Posters should be mounted Monday, October 18, on the designated boards in Conference Hall II and removed at the closing of the conference. The dimensions of the boards are 90 cm wide x 120 cm high. Adhesive tape for mounting the posters on the boards will be available at the poster area. Odd number posters will be viewed at session 1. Even number posters will be viewed at session 2. Posters will be on display for the entire duration of the meeting. (#29 on Hotel Map)


Oral Presentations

Speakers are asked to bring their USB flash drive or CD to the conference reception desk at least 30 minutes prior to the session.  The allocated time for all presentations, unless otherwise indicated, includes five minutes Q&A.


Tour to Knossos and Museum

Buses for the city & archaeological tour will be leaving from the hotel on Tuesday, October 19 at 2:00p.m. Participants will be picked up at the hotel front lobby at that time. Badges are required for participation.


Breakfast for registered participants and registered accompanying persons will be served at the Aldemar Knossos Royal Village hotel main restaurant during the hours scheduled in this program. (#2 on Hotel Map)


Lunches/Informal Discussions

Lunches for registered participants and registered accompanying persons will be served during the times indicated in this program at the Main Restaurant. Badges are required for admission. (#2 on Hotel Map)


Dinners/Informal Discussions

Three of the dinners/informal discussion will be served during the times indicated in the program. Vegetarian entrees are available only upon request in advance of the meeting.  Please contact


Internet, Phone, & Computer

Wireless Internet access is located throughout the conference center free of charge during meeting hours. There are computers available in the hotelÕs business center for the use of the attendees. Phone service is available for an additional charge from your hotel room.


Other Information

Animals are not permitted on site.

Badges are required for admission to all events.

All Aegean Conferences events are non-smoking.


No Photographs and Recording Devices

Participants are not allowed to photograph and/or record using cameras, mobile telephones and other recording devices during the slide and poster presentations.


Tour and Travel Information

A desk operated by the personnel of our official travel agency


Conferex LLC, P.O. Box 12, Wynnewood, PA 19096, USA

Tel.: 1-484-685-0558, FAX: 1-610-771-4224



ERA Ltd., 17, Asklipiou Str., 106 80 Athens-Greece

Tel.: 30-210-363-4944, FAX: 30-210-3631690, E-Mail:

will be located at the conference center throughout the meeting.





Saturday, October 16, 2010

2:00 PM – 6:00 PM

Secretariat Meeting

Theater Room Hotel Lobby


Sunday, October 17, 2010

7:00 AM-8:00 AM




8:00 AM-9:00 AM

Registration: Bioinformatics Workshop

Lobby I

9:00 AM-12:30 PM

Bioinformatics Workshop (see appendix 1)

Tutorial 1 Room Orpheas

Tutorial 2 Room Apollon

Tutorial 3 Room Hermes



12:30 PM-1:00 PM

Coffee Break



11:00 AM

Registration: Student Satellite Symposium

Lobby I

1:00 PM-2:45 PM

Student Satellite Symposium Session 1

Conference Hall I



2:45 PM-3:30 PM

Coffee Break



3:30 PM-5:30 PM

Student Satellite Symposium Session 2

Conference Hall I



4:00 PM – 7:00 PM

24th IMGS Registration Open

Lobby I



7:00 PM-10:30 PM

Welcome Reception and Dinner


Monday, October 18, 2010

7:00 AM-8:20 AM




8:00 AM-5:30 PM

24th IMGS Registration Open

Lobby I



8:20 AM-8:30 AM

Official 24th IMGS Opening



8:30 AM-10:30 AM

Session 1:  Infection and Immunity

Conference Hall I



10:30 AM–12:30 PM

Coffee Break/ Exhibition and Poster Session 1

Odd Numbered Posters

Conference Hall II



12:00 PM-1:30 PM

Editorial Board Meeting with lunch

Theater Room Hotel Lobby



12:30-1:30 PM




1:30 PM-3:30 PM

Session 2: Modeling Disease-Development, Metabolism and Physiology

Conference Hall I



3:30 PM-4:00 PM

Coffee Break



4:00 PM-5:00 PM

IMGS Business meeting (all welcome)

Conference Hall I



5:00 PM-6:00 PM

Verne Chapman Lecture

Conference Hall I



6:30 PM-9:30 PM



Tuesday, October 19, 2010

7:00 AM-8:30 AM

Breakfast and Mentor Breakfast



8:30 AM-10:00 AM

Session 3: Neuroscience, Behavior, and Sensory Systems I

Conference Hall I



10:00 AM

Session 4: Large Scale and Genome-wide Resources

Conference Hall I




Coffee Break


11:00 AM

Session 4 continued: Large Scale and Genome-wide Resources


1:00 PM-2:00 PM




2:00 PM-5:00 PM

Guided Tour of the Knossos Archeological Site

Buses depart hotel lobby at 2:00 PM



5:00 PM-6:30 PM


Human Disease and Mouse Models

Conference Hall I



6:30 PM-9:30 PM



Wednesday, October 20, 2010

7:00 AM-8:30 AM




8:30 AM-10:30AM

Session 5:

Epigenetics, Neoplasia and Aging I

Sponsored by The Ellison Medical Foundation

Conference Hall I




10:30 AM-12:30 PM

Coffee Break/Exhibition and Poster Session 2

Even Numbered Posters

Conference Hall II



12:30 PM-1:30 PM


12:30 PM-1:30 PM

Secretariat meeting with lunch

Theater Room Hotel Lobby



1:45 PM-3:15 PM

Session 6:

Epigenetics, Neoplasia and Aging II

Sponsored by The Ellison Medical Foundation

Conference Hall I




3:15 PM-3:45 PM

Coffee Break



3:45 PM-4:30 PM

Session 7: Neuroscience, Behavior and Sensory Systems II

Conference Hall I



7:00 PM

Farewell Dinner

























Saturday, October 16, 2010

2:00 PM – 6:00 PM

Secretariat Meeting

Theater Room Hotel Lobby


Sunday, October 17, 2010

7:00 AM-9:00 AM





8:00 AM-9:00 AM

Registration: Bioinformatics Workshop

Lobby I

9:00 AM-12:30 PM

Bioinformatics Workshop (see appendix 1)

Tutorial 1 Room Orpheas

Tutorial 2 Room Apollon

Tutorial 3 Room Hermes



12:30 PM-1:00 PM

Coffee Break


11:00 AM


Registration: Student Satellite Symposium

Lobby 1


Student Satellite Symposium Session 1

Conference Hall I




1:00 PM


The F2 population for dissecting host susceptibility to periodontitis

Ariel Shusterman, Yael Houri-Haddad, Ervin  I Weiss, and Fuad A Iraqi

1:15 PM


Mouse genetic model of cardiovirulent coxsackievirus B3 infection

Sean A. Wiltshire and Silvia M. Vidal

1:30 PM


Association between spatial proximity and functional similarity in human genome

Ekaterina Khraneeva, Andrey Mironov, and Mikhail Gelfand

1:45 PM


Olfactory receptor gene family evolution in marsupials

Amir Mohammadi, Margaret L. Delbridge, and Jennifer A. Marshal Graves

2:00 PM


Advancing paternal age is associated with deficits in social and exploratory behaviours in the offspring

Rebecca G. Smith, Rachel L. Kember, Leonard C. Schalkwyk, Joseph D Buxbaum, Abraham Reichenberg, Cathy Fernandes, and Jonathan Mill

2:15 PM


Genome-wide differential methylation patterns in intersubspecific hybrid mice

John D. Calaway, Hyuna Yang, Elena de la Casa-Esperon, Megan E. Hanson, Ezequiel C. Cambranis, Maria A. Perez-Lamigueiro, David L. Aylor, Leonard McMillan, Gary A. Churchill, and Fernando Pardo-Manuel de Villena

2:30 PM


Optimization of a genome-wide, allele-specific DNA methylation assay for the mouse

John P. Didion, John D. Calaway, Isa Kemal Pakatci, Leonard McMillan, and Fernando Pardo-Manuel de Villena




2:45 PM

Coffee Break




Student Satellite Symposium Session 2

Conference Hall I




3:30 PM


Characterisation of laterality and cardiac developmental defects in the K27 mutant mouse

Louise Stephen, Karen Mitchell, and Kathryn Hentges

3:45 PM


Kyoto Epileptic Rat (KER): A novel rat model of Kcna1 mutation showing spontaneous seizures and muscle involuntry contractions

Saeko Ishida, Tomoji Mashimo, Takeshi Nishio, Yu Sakamoto, Shuji Kaneko, and Tadao Serikawa

4:00 PM


Identifying modifier genes of mpnsts in the Nf1;p53cis mouse model of neurofibromatosis type 1

Jessica A Van Schaick, Keiko Akagi, Sandra Burkett, Christina DiFabio, Robert Tuskan, Jessica Walrath, and Karlyne Reilly

4:15 PM


A novel genetic model of osteoporosis by overexpression of human RANKL in transgenic mice

Alexandra Niti, R Dacquin, V Rinotas, P Jurdic, and E Douni

4:30 PM


Positive selection might contribute to functional divergence of NAIPs following gene duplication in mammals during the evolution

Zhaoliang Su, Chenglin Zhou, Jianguo Chen, Shengjun Wang, and Huaxi Xu

4:45 PM


A forward genetics approach identifies a novel member of the Dnajc family that causes neuromuscular disease in mice

Fotios Ioakeimidis, Vagelis Rinotas, Eleni Makrinou, George Kollias, and Eleni Douni

5:00 PM


Gene expression studies of ncRNAs within Odz4 locus

Tyler Buit, Chiao-Ling Lo, Feichen Shen, and Amy C. Lossie

5:15 PM


Early life stress is associated with altered behavior and epigenetic alterations to Nr3c1, Avp and Nr4a1

Rachel L. Kember, Emma Dempster, Leonard C. Schalkwyk, Jonathan Mill and Cathy Fernandes




4:00 PM -7:00 PM

24th IMGS Registration Open

Lobby I



7:00 PM-10:30 PM

Welcome Reception and Dinner




Monday, October 18, 2010

7:00 AM-8:20 AM





8:00 AM-5:30 PM

24th IMGS Registration Open

Lobby I




8:20 AM

Official 24th IMGS Opening




Session 1: Infection and Immunity

Conference Hall I

8:30 AM


Stress mechanisms, inflammation, infection and host resistance

Katia P. Karalis

9:15 AM


Physiologic consequences of constitutive MTOR inhibition: alterations in cell/organ size and immune cell development

Shuling Zhang, Julie A. Readinger, Wendy DuBois, Mirkka Janka-Junttila, Richard Robinson, Magaret Pruitt, Val Bliskovsky, Julie Z. Wu, Kaori Sakakibara, Jyoti Patel, Carole A. Parent, Lino Tessarollo, Pamela L. Schwartzberg, and Beverly A. Mock

9:30 AM


Severe Combined Immunodeficiency (SCID) rats generated by zinc finger nuclease technology

Tomoji Mashimo, Akiko Takizawa, Birger Voigt, Kazuto Yoshimi, Hiroshi Hiai, Takashi Kuramoto, and Tadao Serikawa

9:45 AM


A recombination hotspot leads to sequence variability within a novel gene and contributes to complex disease susceptibility

Iris KL Tan, Leanne Mackin, Nancy Wang, Anthony T Papenfuss, Colleen M Elso, Michelle P Ashton, Belinda Phipson, Melanie Bahlo, Terrence P Speed, Gordon K Smyth, Grant Morahan, and Thomas C Brodnicki

10:00 AM


Selected Student Talk

10:15 AM


Selected Student Talk




10:30 AM

Coffee Break/ Exhibition and Poster Session 1

Odd Numbered Posters

Conference Hall II




12:30 PM-1:30 PM




12:00PM-1:30 PM

Mammalian Genome Editorial Board Meeting with Lunch

Theater Room Hotel Lobby




Session 2: Modeling Disease: Development, Metabolism and Physiology

Conference Hall I


1:30 PM


A sensitized suppressor screen to identify modifier genes for factor V leiden-dependent thrombosis in the mouse

Randal Westrick, Goujing Zhu, Sara Manning, Angela Yang, David Siemieniak, and David Ginsburg

1:45 PM


Genetic and dietary controls of alternative exon use and mRNA levels in the systems architecture of diet-induced metabolic diseases

Sabrina H. Spiezio, Annie Hill-Baskin, Karen Fitch, Keith Jones, Michael H. Shapero, and Joseph H. Nadeau

2:00 PM


A gene driven ENU mutagenesis screen reveals a novel method of ZIC protein nuclear localization

R Ali, H Bellchambers, N Warr, D Quwailid, P Denny and Ruth Arkell

2:15 PM


Non-muscle myosin IIB is required for formation of the coronary vasculature

Karen Mitchell, C Clowes, Louise Stephen, and Kathryn E Hentges

2:30 PM


Short circuit, a homozygous lethal ENU mutation that affects lung development

Michael J. Parsons, Laura Yates, Jessica Edwards, Lauren Chessum, Charlotte H. Dean, and Patrick M. Nolan

2:45 PM


Identification of the DanforthÕs short tail mutation using next generation sequencing

Christopher N. Vlangos, Amanda N. Siuniak, Dan Robinson, Arul M. Chinnaiyan, James Cavalcoli, Robert H. Lyons, and Catherine E. Keegan

3:00 PM


A novel role for Atmin, controlling ciliogenesis through modulation of dynein light chain expression

Paraskevi Goggolidou, Jonathan Stevens, Gabrielle Wheway, Antonella Di Paolo, Rosario Romero, James Briscoe, Colin Johnson, and Dominic Norris

3:15 PM


Genetic analysis of complex traits in the emerging collaborative cross

David L. Aylor, William Valdar,, Wendy Foulds-Mathes, Ryan J. Buus, Ricardo A. Verdugo,, Ralph S. Baric, Martin T. Ferris, Jeffrey A. Frelinger, Mark Heise, Matt B. Frieman, Lisa E. Gralinski, Timothy A. Bell, John P. Didion, Kunjie Hua, Derrick L. Nehrenberg, Christine L. Powell, Jill Steigerwalt, Yuying Xie, Samir N.P. Kelada, Francis S. Collins, Ivana V. Yang, David A. Schwartz, Lisa A. Branstetter, Elissa J. Chesler, Darla R. Miller, Jason Spence, Eric Yi Liu, Leonard McMillan, Abhishek Sarkar, Jeremy Wang, Wei Wang, Qi Zhang, Karl W. Broman, Ron Korstanje, Caroline Durrant, Richard Mott, Fuad A. Iraqi, Daniel Pomp, David Threadgill, Fernando Pardo-Manuel de Villena, and Gary A. Churchill




3:30 PM-4:00 PM

Coffee Break




4:00 PM-5:00 PM

IMGS Business meeting (all welcome)

Conference Hall I




5:00 PM –

6:00 PM


Verne Chapman Lecture

Realising the potential of mouse disease models – from pathway to therapy

Steve D.M. Brown

Conference Hall I




6:30 PM-9:30 PM



Tuesday, October 19, 2010

7:00 AM-8:30 AM

Breakfast and Mentor Breakfast




Session 3: Neuroscience, Behavior, and Sensory Systems I

Conference Hall I




8:30 AM


Importance of Cadm1 and cell adhesion in depressive behavior

C Santos, B Miller, M Pletcher, A Su, L Tarantino, and Tim Wiltshire

8:45 AM


An ENU sensitization screen to determine the physiological function of a- synuclein

Deborah E Cabin, M Casey, and D Zou

9:00 AM


Genomic resources in Peromyscus – mapping the audiogenic seizure sensitivity locus

Gabor Szalai, Janet Crossland, Jay Coleman, and Michael Felder

9:15 AM


Mutations in the PDZ domain containing protein Gipc3 cause progressive sensorineural degeneration (Ahl5 and jams1) in mice and recessive hearing impairment in humans (DFNB95)

Nikoletta Charizopoulou, Andrea Lelli, Margit Schraders, Kausik Ray, Ronald J.C. Admiraal, Harold R. Neely, Joseph R. Latoche, John K. Northup, Hannie Kremer, Jeffrey R. Holt, and Konrad Noben-Trauth

9:30 AM


A mutation in the gene encoding mitochondrial Mg2+ channel MRS2 results in demyelination in rats

Takashi Kuramoto, Mitsuru Kuwamura, Satoko Tokuda, Takeshi Izawa, Yoshifumi Nakane, Kazuhiro Kitada, Masaharu Akao, Jean-Louis GuŽnet, and Tadao Serikawa

9:45 AM


Selected Student Talk




Session 4: Large Scale and Genome-wide Resources

Conference Hall I


10:00 AM


Annotation of long non-coding RNA transcripts

Mark Thomas and Jennifer Harrow

10:15 AM


The JAX Cre Repository: Improving the utility of Cre driver strains

Stephen A. Murray, Caleb Heffner, Michael Sasner, Cathleen Lutz, Brandon Grossman, Stephen Rockwood, Yashoda Sharma, and Leah Rae Donahue



10:30 AM

Coffee Break




11:00 AM


Mouse genomes project: map of SNPS and short indels

Petr Danecek, Gan Xiangchao, Thomas M Keane, James Stalker, Binnaz Yalcin, Martin Goodson, Sendu Balasubramaniam, Kim Wong, Guy Slater, Andreas Heger, Eleazar Eskin, Nick Furlotte, Chris Ponting, Jonathan Flint, and David J Adams

11:15 AM


Use of PiggyBac-mediated transient transgenic RNAi expression for rapid characterization of gene function during embryonic development

David R. Beier, Yuko Fujiwara, Shannon W. Davis, Haiyan Qiu, Thomas L. Saunders, Stuart Orkin, Sally A. Camper, and Bryan C. Bjork

11:30 AM


The Sanger Institute Mouse Genetics Programme

Ramiro Ramirez-Solis, J White, E Ryder, R Houghton, and J Bottomley

11:45 AM


Next level systemic phenotyping of mice

Martin HrabŽ de Angelis and the German Mouse Clinic consortium

12:00 Noon


Portal to large-scale mouse knockout resources

William Skarnes

12:15 PM


Tooling up MGI for a deluge in mouse phenotype and disease model data

Anna Anagnostopoulos, Janan Eppig, and Mouse Genome Informatics Group

12:30 PM


New approaches to conditionality

Aris N. Economides, David Frendewey, Peter Yang, David M. Valenzuela, Andrew J. Murphy, and George D. Yancopoulos

12:45 PM


Mouse metabolic phenotyping centers:  National consortium focusing on diabetic complications

Renee LeBoeuf




1:00 PM-2:00 PM





2:00 PM-5:00 PM

Guided Tour of the Knossos Archeological Site

Buses depart hotel lobby at 2:00 PM




5:00 PM-6:30 PM


Human Disease and Mouse Models

Conference Hall I

5:00 PM



Edward Wakeland

5:45 PM


Cancerous microRNAs and regulatory RNA binding proteins

Reuven Agami




6:30 PM





Wednesday, October 20, 2010

7:00 AM-8:30 AM





Session 5: Epigenetics, Neoplasia and Aging I

Sponsored by The Ellison Medical Foundation

Conference Hall I

8:30 AM


Genomic imprinting: Insights from the Gnas cluster 

Jo Peters

9:15 AM


The battle of the signals: Non-coding RNAs vs. enhancers in the transcriptional regulation of the Kcnq1 imprinted region

Anjali Raval, Lisa Korostowski, and Nora Engel

9:30 AM


An identity crisis: Abnormal male germ cell development and the initiation of testicular cancer

Jason Heaney, Jean Kawasoe, Megan Michelson, and Joseph Nadeau

9:45 AM


Age-associated change of energy metabolism are genetically dissected by mouse inter-subspecific consomic strains

Toyoyuki Takada, Akihiko Mita, Shigeharu Wakana, Kazuo Moriwaki, Hiromichi Yonekawa, and Toshihiko Shiroishi

10:00 AM


Genetic regulation of aging related phenotypes and lifespan in mice

Rong Yuan, S W Tsaih, Q Meng, K Flurkey, J Nautiya, SB Petkova, MA Bogue, KD Mills, L L Peters, CJ Bult, CJ Rosen, JP Sundberg, M Parker, DE Harrison, GA Churchill, and B Paigen

10:15 AM


Subspecific origin and haplotype diversity in the laboratory mouse

Hyuna Yang, Jeremy R Wang, John P Didion, Ryan J Buus, Timothy A Bell, Catherine E Welsh2, Franois Bonhomme, Alex Hon-Tsen Yu, Michael W Nachman, Jaroslav Pialek, Priscilla Tucker, Pierre Boursot, Leonard McMillan, Gary A Churchill, and Fernando Pardo-Manuel de Villena




10:30 AM

Coffee Break/Exhibition and Poster Session 2

Even Numbered Posters

Conference Hall II




12:30 PM-1:30 PM




12:00 PM-1:30 PM

Secretariat meeting with lunch

Theater Room Hotel Lobby



Session 6: Epigenetics, Neoplasia and Aging II

Conference Hall I



Sponsored by The Ellison Medical Foundation

1:45 PM


David Galas

2:30 PM


Genetic architecture of hybrid sterility: Matching the puzzle pieces together

Jiri Forejt, Maria Dzur-Gejdosova, Vaclav John, Tanmoy Bhattacharyya, Petr Simecek, Sona Gregorova, Ondrej Mihola, Petr Flachs, and Zdenek Trachtulec

2:45 PM


Molecular interactions of dead end (Dnd1)

Angabin Matin, Zhu Rui, KangLi Luo, Chitralekha Bhattacharya, Michelina Iacovino, Elisabeth Mahen, Michael Kyba, and Sita Aggarwal

3:00 PM


Genetic dissection of metatasis susceptibility converges on a common mechanism

J Alsarraj, S Winter, N Goldberger, K Mattaini, M William, L Lukes, R Walker, and Kent W Hunter




3:15 PM


Coffee Break




Session 7: Neuroscience, Behaviour and Sensory Systems II

Conference Hall I

3:45 PM


Icst is a dominant negative mutation of lmx1b

Sally H. Cross, Lisa Mckie, Margaret Keighren, Dan Macalinao, Alison L. Kearney, Simon W. John, and Ian J. Jackson

4:00 PM


Elucidating the role of Nsdhl and cholesterol metabolism in CNS development using a conditional knockout allele

Gail E Herman, N Bir, L Binkley, K McLarren, C Boerkoel, and D Cunningham

4:15 PM


Genetic, stem cell, and systems analyses of neurodegenerative diseases

George A Carlson, R Bennett, ME Orr, I Lee, H Yoo, J-H Cho, D Hwang, and LE Hood




7:00 PM


Farewell Dinner







Stage and cell subtype-specific epigenetic regulation of mammary gland development

Deanna Acosta, Melissa J Fazzari, John M Greally, and Cristina Montagna


Toward an unified measure of intraspecific selective pressure

Roberto Amato, Gennaro Miele, Michele Pinelli, and Sergio Cocozza


A new targeted mutation, Caspa, in the Gnas complex shows hyperactivity and ataxia

Simon Ball, Christine Williamson, Charlotte Tibbit, and Jo Peters


Rat resource and research center

Beth A. Bauer, EC Bryda, CL Franklin, LK Riley, and JK Critser


Loci on chromosome 2 are associated with na•ve airway hyperresponsiveness in CD-1 outbred mice

David R Beier, Andrew Kirby, Mary Prysak, Mark Daly, and Emily Cozzi


High-resolution map and candidate gene analysis for ŽbouriffŽ (ebo), a hair mutation on mouse Chromosome 2 associated with infertility

Fernando Benavides, J Jaubert, CJ Perez, J-L GuŽnet, J Barrera, B Cole, and CJ Conti


Molecular genetic analysis of the NADPH oxidase of the vestibular system

Catrina A Spruce, John P Flaherty, Heather E Fairfield and David E Bergstrom


Mouse models of Costello syndrome

Heather E Fairfield, John P Flaherty, Catrina A Spruce, Leah Rae Donahue and David E Bergstrom


The application of high-throughput sequencing technologies for mutation discovery in the Jackson LaboratoryÕs mouse mutant resource

Laura G Reinholdt, David E Bergstrom, Muriel T Davisson-Fahey, Cathleen M Lutz, Michael Sasner, Steven A Murray, Jeff Lake, Steven Rockwood, Leah Rae Donahue and the MMR team


The role of CAML in the inner ear

Elizabeth C Bryda, Nathan T Johnson, Cynthia L Besch-Williford, Kevin K Ohlemiller, and Richard J Bram


Interesting phenotypes found as part of the infection challenge in the Wellcome Trust Sanger InstituteÕs Mouse Genetics Programme

Simon Clare, Leanne Kane, Lynda Mottram, Jacqui White, Ramiro Ramirez – Solis, and Gordon Dougan


The expression of SEPTIN 9 in human breast cancer

Diana Connolly, Zhixia Yang, Elias T. Spiliotis, Maria Castaldi, Nichelle Simmons, Pascal Verdier-Pinard, and Cristina Montagna


Genetic contribution to liver fibrosis

David DeSantis, Michelle Pritchard, Laura Nagy, Joseph Nadeau, and Colleen M Croniger


Diet, obesity, and susceptibility to colon polyps

Stephanie K. Doerner and Joseph H. Nadeau


Genetic regulation of fracture risk in inbred mice

Leah Rae Donahue GA Churchill, S-W Tsaih, and WG Beamer


Imprinted XLAS: A new player in bone and adipocyte metabolism

Sally Eaton, Simon Ball,  Colin Beechey, Christine Williamson, and Jo Peters


The Sanger Mouse Genetics Programme: High throughput recessive lethality screen

Jeanne Estabel, Elizabeth Tuck, Damian Carragher, Jennifer Salisbury, and Jacqueline K. White on behalf of the Mouse Genetics Programme


Multiple forebrain cis-regulatory elements at the Dlx1/2 locus: redundant function or complex control mechanisms?

Marc Ekker, Luc Poitras, Man Yu, Noel Ghanem, and Lisa Tran


Novel mutation discovery on ENU mouse mutagenesis by next-generation sequencer

Ryutaro Fukumura, Hayato Kotaki, Yuichi Ishitsuka, Takuya Murata, Shigeru Makino, Yuji Nakai, and Yoichi Gondo


The Sanger Mouse Genetics Programme: High throughput characterisation of knockout mice for genes involved in glucose metabolism

Anna-Karin Gerdin, L Roberson, Jacqueline K White on behalf of the Mouse Genetics Programme


EuroPhenome: Large dataset visualisation, statistical analysis and data exploratory tool for mouse phenotyping data

Ahmad Hassan, Hugh Morgan, Andrew Blake, Simon Greenaway, The EUMODIC Consortium, John M. Hancock, and Ann-Marie Mallon


The intragene resource centre, a national centre for mouse genetics, distribution, archiving and functional imaging

Cecile Fremond, Stephanie Lerondel, Christelle Martin, and Yann Herault


The Down syndrome as a complex trait resulting from the interaction of dosage sensitive genes unravelled in the mouse model

V Brault, A Duchon, E Dalloneau, P Lopes, I Abizanda, S Luiller, JC Bizot, C Borel, M Raveau, D Marechal, J Lignon, S Pothion, F Trovero, V Tybulewicz, EM Fisher, S Antonarakis, M Dierssen, and Yann Herault


The European Mouse Mutant Archive (EMMA)

Michael Hagn, G Tocchini-Valentini, Y HŽrault, S Brown, U Lendahl, J Demengeot, M HrabŽ de Angelis (EMMA Director), E Birney, R Ramirez-Solis, JL Mandel, and L Montoliu


Broad based phenotyping approaches for the characterisation of mutant mouse lines in EUMODIC and the Harwell aging screen

Heena Vanmalibhai Lad, KA Lee, A Blake, H Morgan, A Hassan, S Greenaway, AM Mallon, HJ Gates, SDM Brown, and The EUMODIC Consortium


FBXO11 regulates TGF beta signaling through TRP53

Hilda Tateossian, Susan Morse, and Steve DM Brown


The development of the integrated database of mammals in RIKEN

Hiroshi Masuya, Yuko Makita, Koro Nishikata, Yuko Yoshida, Terue Takatsuki, Kasunori Waki, Nobuhiko Tanaka, Yoshiki Mochizuki, Norio Kobayashi, Riichiro Mizoguchi, Teiichi Furuichi, Hideya Kawaji, Daub Carsten, Yoshihide Hayashizaki, Shigeharu Wakana, Atsushi Yoshiki, Kaoru Fukami-Kobayashi, and Tetsuro Toyoda


Combining clinical and expression QTLs in the context of infection: new insights into susceptibility to influenza

Gregory A. Boivin, Julien Pothlichet, Emil Skamene, Earl G. Brown, Robert Sladek, and Silvia M. Vidal


A nature of the blastomer DNA methylation determination

Andrey A. Ivanov


Not all wild-derived Mus spretus mice do resist virulent Yersinia pestis

Jean Jaubert, Charlotte Leblanc, Charlne Blanchet, Garcia Sylvie, Elisabeth Carniel, Christian Demeure, Jean-Jacques Panthier, and Xavier Montagutelli


Evidence of hypothalamic degeneration in the anx/anx mouse

IAK Nilsson, S Thams, C Lindfors, A Bergstrand, S Cullheim, T Hškfelt, and Jeanette E Johansen


Identification of a novel Slc25 member that causes autosomal recessive ataxia in ENU-mutagenized mice

A Karakostas, F Ioakeimidis, V Rinotas, G Kollias, and Eleni Douni


Consomic analysis of genetic factors related to temporal difference of home-cage activity between B6 and MSM

Ayako Ishii, Akinori Nishi, Toshihiko Shiroishi, Aki Takahashi, and Tsuyoshi Koide


Genetic determinants for intramuscular fat content and water holding capacity in mice selected for high muscle mass

Stefan KŠrst and Gudrun A. Brockman


Discovering novel regulatory pathways of imflammation by ENU mutagenesis

Philip L Kong, Emma Timms, Andrew Coulson, Kristina Blagoeva-Hubbard, Paul Potter, and Marc Feldmann


Epigenetic changes in Friedreich ataxia pathogenesis and therapy

S Al-Mahdawi, C Sandi, R Mouro-Pinto, V Ezzatizadeh, and Mark A Pook


A mouse mutant exhibiting a unique pattern of cochlear inner hair cell degeneration

Morag A. Lewis, Tracy J. Bussoli, Amy Taylor, Michael A. Cheong, Cordelia F. Langford, and Karen P. Steel


A comprehensive whole-genome map of endogenous retroviral elements and their functional effects across 17 laboratory mouse strains

Thomas M Keane, K Wong, J Flint, W Frankel, David J Adams


Phenotyping of knockout mice using bacterial pathogens as part of the Wellcome Trust Sanger InstituteÕs Mouse Genetics Programme

Leanne Kane, Lynda Mottram, Simon Clare, Jacqui White, Ramiro Ramirez – Solis, and Gordon Dougan


Imprinted methylation vs. imprinted expression – are they necessarily associated?

John D. Calaway, Hyuna Yang, Megan E. Hanson, Ezequiel C. Cambranis, Maria A. Perez-Lamigueiro, David L. Aylor, Leonard McMillan, Gary A. Churchill, Fernando Pardo-Manuel de Villena, and Elena de la Casa-Esperon


Dual activity on a single locus: Existence of a novel RNA control mechanism for microRNA-650 and immunoglobulin lambda light chain variable genes in primates

Sabyasachi Das, Jianxu Li, Masayuki Hirano, Christopher L. Haga, Murali Gururajan, Gštz R.A. Ehrhardt, and Max D. Cooper


Identification of positional candidates for Chr15 obesity quantitative trait loci using microarray and RT-PCR analysis of F2 congenic crosses in mice

K Cirnski, M Stanonik, R Keber, P Juvan, R Košir, D Rozman, and Simon Horvat


Crezoo - the european virtual repository for cre driver strains

Christina Chandras, Michael Zouberakis, Damian Smedley, Nadia Rosenthal, and Vassilis Aidinis


Host immune responses and genetic factors modulating resistance to Salmonella enterica serovar typhimurium in the inbred mouse strain SPRET/Ei: a role for neutrophils

Lien Dejager, Iris Pinheiro, Pieter Bogaert, Liesbeth Huys, and Claude Libert


LPS-resistance of the mouse strain SPRET/Ei: an important role of the X-chromosome

Iris Pinheiro, Lien Dejager, Tina Mahieu, Marnik Vuylsteke, and Claude Libert


Complete knockout of the adrenocortical dysplasia gene encoding the shelterin protein TPP1 is associated with telomere dysfunction and early embryonic lethality

Gail A Osawa, C Harris, T Kibe, T de Lange, S Kalantry, and CE Keegan


The FaceBase Cre driver project: creating new mouse tool strains for clefting research

Stephen A. Murray, Jocelyn Sharp, John Flaherty, Thomas Gridley, and Leah Rae Donahue


Epigenome sequencing comes of age in development, differentiation and immune regulation mechanisms of mammalian research

Ning Li and Jun Wang


Generation of customized and read-to-use genetically engineered mice at the institute Clinique de la souris – ICS (The French Mouse Clinical Institute)

Marie-Christine Birling, AndrŽe Dierich, Karim Essabri, Lydie Venteo, Guillaume Pavlovic, Tania Sorg, and Yann HŽrault


Pudding – a novel model of glomerularnephritis

Paul K Potter, L Wisby, FW Tam, S Wells, T Hough, HT Cook, M Cheeseman, and SDM Brown


The Harwell ageing screen

Paul K Potter


Gelsolin plays a role in the actin polymerization complex of hair cell stereocilia

Philomena Mburu, MR Romero, Helen Hilton, Andrew Parker, Stuart Townsend, Yoshiaki Kikkawa, and SDM Brown


New sterile model mouse from RIKEN ENU-based gene-driven mutagenesis

Takuya Murata, Shigeru Makino, Ryutaro Fukumura, and Yoichi Gondo


Genetic diversity among C57BL/6 substrains based on SNPs

Kazuyuki Mekada, Ikuo Miura, Takehide Murata, Atsushi Toyoda, Mayu Hirose, Shigeharu Wakana, Kuniya Abe, and Atsushi Yoshiki


Novel ENU screens for immune-related phenotypes

Vera M Ripoll, Marie Hutchison, Paul Potter, and Roger Cox


Grb10 modifies MPNST tumorigenesis in a mouse model of NF1

Karlyne M. Reilly, Jessica Van Schaick, Keiko Akagi, Christina DiFabio, Robert Tuskan, Sandra Burkett, and Jessica Walrath


Homology mapping: development of the mouse hindlimb musculoskeletal model using the human musculoskeletal model and Scleraxis (Scx)-GFP mouse

Satoshi Oota, Nobunori Kakusho, Yosuke Ikegami, Kazuyuki Mekada, Koh Ayusawa, Hirotaka Imagawa, Yuichi Obata, Ryutaro Himeno, Hideo Yokota, Yoshihiko Nakamura, and Atsushi Yoshiki


Mutation identification in mice by exome sequencing

Laura Reinholdt, David Bergstrom, Steve Murray, Michelle Curtain, Carol Bult, Joel Richardson, Lucy Rowe, Mary Barter, Daniel J. Gerhardt, Mark D'Ascenzo, Todd Richmond, Tom J. Albert, Jeffrey A. Jeddeloh, Benjamin Kile, Ivo Gut, Jay Shendure, and Leah Rae Donahue


Status of the US lines of the Collaborative Cross: Completed lines

Darla R Miller, Ryan J Buus, Jennifer N Shockley, Stephanie D Hansen, Ginger D Shaw, Terry J Gooch, Jason S Spence, Timothy A Bell, Catherine E Welsh, Leonard McMillan, Wei Wang, Jeremy Wang, Eric Yi Liu, Kenneth F Manly, Gary A Churchill, David W Threadgill, and Fernando Pardo-Manuel de Villena


Whole genome resequencing of Estonian dairy cow (Estonian Holstein)

Rutt Lilleoja, † Jaakma, and S K›ks


Hypothalamic mitochondrial dysfunction, a possible cause of anorexia

Charlotte Lindfors, Ida A.K. Nilsson, Pablo M. Garcia-Roves, Aamir R. Zuberi, Mohsen Karimi, Lea Rae Donahue, Derry C. Roopenian, Jan Mulder, Mathias UhlŽn, Tomas J. Ekstršm, Muriel T. Davisson, Tomas Hškfelt, Martin Schalling, and Jeanette E. Johansen


High-throughput genotyping and quality control of gene-targeted mutations in the mouse

Edward Ryder, Diane Gleeson, Debarati Bhattacharjee, Ross Cook and the Mouse Genetics Programme


The Sanger Mouse Genetics Programme: High throughput characterisation of immunological phenotypes in knockout mice

Mark Lucas, C Podrini, E Cambridge, S Clare, L Mottram, JK White on behalf of the Mouse Genetics Programme


The Mouse Reference GENOME assembly: updates and resources

Valerie A Schneider and DM Church on behalf of the Genome Reference Consortium and NCBI Genome Annotation Team


dbVar and dbSNP: NCBI Databases of Simple and Structural Variations

Valerie A Schneider, Tim Hefferon, John Garner, Azat Mardanov, Ming Ward, Aleksey Vinokurov, Melvin Quintos, Mike Kholodov, David Shao, John Lopez, Steve Sherry, and Deanna Church


Current progress of phenotypic analysis in Japan mouse clinic

Tomohiro Suzuki, Hideki Kaneda, Kimio Kobayashi, Ikuo Miura, Tamio Furuse, Ikuko Yamada, Osamu Minowa, Hideaki Toki, Hiromi Motegi, Maki Inoue, Tetsuo Noda, Hiroshi Maduya, and Sigeharu Wakana


Hyperactive mice show a cluster of associated metabolic features

Jacqueline K White, Anna-Karin Gerdin, Christine Podrini on behalf of the Mouse Genetics Programme, and Karen P Steel


Multiplicity of phenotypes in early generations of the mouse diversity outcross population

Karen L Svenson and GA Churchill


SDOP-DB: a database for international sharing and standardization of mouse phenotyping protocols

Nobuhiko Tanaka, Kazunori Waki, Hideki Kaneda, Tomohiro Suzuki, Ikuko Yamada, Tamio Furuse, Kimio Kobayashi, Hiromi Motegi, Hideaki Toki, Maki Inoue, Osamu Minowa, Tetsuo Noda, Shigeharu Wakana, and Hiroshi Masuya


Modeling human chromosome 21-associated mental retardation disorders in mice

Tao Yu, Zhongyou Li, Steven J Clapcote, Zhengping Jia, Chunhong Liu, Annie Pao, Sei-ichi Matsui, Norma J Nowak, John C Roder, Chu Chen, Allan Bradley, and Y. Eugene Yu


Taiwan Mouse Clinic – an experiment for mouse phenotyping service

Jeffrey Jong-Young Yen, Yen-Hui Chen, and Ya-Wen Hsaio


Initial assembly of 17 mouse strains from the mouse genomes project

Guy Slater, Thomas M Keane, Jared Simpson, Aylwyn Scally, Xiangchao Gan, Richard Mott, Jonathan Flint, and David J Adams


Clustered protocadherin genes: Genetic codes for generating the complex brain

Takeshi Yagi, Shunsuke Toyoda, Yasushi Itoga, Keizo Hirano, Atsushi Okayama, Shinichi Yokota, Shota Katori, Teruyoshi Hirayama, and Takahiro Hirabayashi


Comprehensive phenotyping of mouse models

Tania Sorg, Marie-France Champy, Elodie Bedu, Roy Combe, Hamid Meziane, and Yann Herault


Targeted and whole exome resequencing of mouse monogenic mutants, quantitative trait loci and cancer models

David R. Beier, Jennifer L. Moran, Evan Mauceli, Snaevar Sigurdsson, Tim Fennell, Lauren Ambrogio, Miriam H. Meisler, John C. Schimenti, Karen Cichowski, Laura Reinholdt, Jane Wilkinson, Stacey B. Gabriel, Federica di Palma, and Kerstin Lindblad-Toh


Dynamic transcriptomes during neural differentiation of human embryonic stem cells revealed by short, long, and paired-end sequencing

Jia Qian Wu, Lukas Habegger, Parinya Noisa, Anna Szekely, Caihong Qiu, Stephen Hutchison, Debasish Raha, Michael Egholm, Haifan Lin, Sherman Weissman, Wei Cui, Mark Gerstein, and Michael Snyder


Improving mouse genome annotation through new technologies

Laurens Wilming, Gary Saunders, Markus Brosch, Gavin Laird, Adam Frankish, and Jennifer Harrow


Genomic alterations in tumors of the Mcm4Chaos3 breast cancer mouse model

Marsha D. Wallace, Ethan Cerami, Stefan Stefanov, Lishuang Shen, and John C. Schimenti


A missense mutation in the novel WD-repeat-containing protein, WDR81, causes Purkinje cell and photoreceptor cell death in the ENU-induced neurological mouse mutant nur5

Maria Traka, Kathleen J. Millen, and Brian Popko


Using whole genome next-generation sequencing as a tool to find ENU-induced mutations

Michelle Simon, S Greenaway, P Denny, A-M Mallon, and JM Hancock


The Sanger Mouse Genetics Programme; High throughput morphological analysis of the eye

Valerie E. Vancollie, J Estabel, D Sunter, VB Mahajan, SH Tsang, MC Naumann, M Mahajan, JK White, on behalf of the Mouse Genetics Programme


A Cellular Genetics Platform to identify pharmacogenetic toxicity pathways

O Suzuki, N Butz, M Pletcher, A Su, B Steffy, D Scoville, A Frick, J Trask, R Thomas, and Tim Wiltshire


Lessons from phenotype of KO mouse, The role of Snur4 on adipogenesis and insulin sensitivity

Jae Hoon Shin, Il Yong Kim, Yo Na Kim, Ji Won Choi, Kyung Jin Roh, Mi Ra Sohn, Cheol soo Choi, Yun Soo Bae, and Je Kyung Seon


Epiblast-ablation of Sox2 expression leads to neurocristopathies in mouse embryos

Nikolaos Mandalos, Marannia Saridaki, Jessica Lea Harper, Anastasia Kotsoni, Aris N. Economides, and Eumorphia Remboutsika


A comprehensive catalogue of structural variation from‭ ‬17‭ ‬inbred mouse strains

Kim Wong,‭ ‬B Yalcin,‭ ‬T Keane,‭ ‬M Goodson,‭ ‬A Agam,‭ ‬C Nellaker,‭ ‬J Stalker,‭ ‬J Flint,‭ ‬and D Adams


Collection and distribution of CRE/FLP-drivers at the Riken Bioresource Center

Atsushi Yoshiki, Kazuyuki Mekada, Hatsumi Nakata, Ayumi Murakami, Masayo Kadota, Fumio Ike, Noriko Hiraiwa, Kaoru Fukami-Kobayashi, Yuichi Obata


Analysis of the expression profile of CRH-POMC system genes in vitiligo skin biopsies

Ene Reimann,, K Kingo, M Karelson, T Salum, E Aunin, P Reemann, K Abram, E Vasar, H Silm, and S K›ks


Effects of a high spontaneous mutation rate in mammalian germline by using mutator mice modified replicative DNA polymerase delta   

Arikuni Uchimura, Yuko Hidaka, Ikuo Miura, Shigeharu Wakana, and Takeshi Yagi


Tooling up MGI for a deluge in mouse phenotype and disease model data

Anna Anagnostopoulos, Janan Eppig, and Mouse Genome Informatics Group

























The F2 population for dissecting host susceptibility to periodontitis

Ariel Shusterman1, Yael Houri-Haddad1, Ervin I Weiss1, and Fuad A Iraqi2

1Department of Prosthodontics Hadassah Medical Center, Israel

2Deptartment of Clinical Microbiology and Immunology, Sackler Faculty of Medicine, Tel Aviv University, Israel


Introduction: Epidemiological studies in human have suggested that susceptibility to periodontal disease is controlled by host genetic factors. Recently, we have shown quantitatively by using microCT that inbred mouse strains respond differently to the infection. Based on these results, we have developed 408 mice of A/JxBALB/cJ F2 resource population, knowing that A/J and BALB/cJ are the resistant and susceptible founder strains for the infection, respectively.


1)   To determine the phenotypic response; i.e. alveolar bone loss, of all progenies of the F2 population.

2)   To perform genome wide search for quantitative trait loci (QTL) associated with host susceptibility to periodontitis using selective genotyping approach.


Materials and Methods: 408 mice of the F2 population were generated by a controlled mating scheme of the two parental inbred founders. All mice were orally infected with Porphyromonas gingivalis and Fusobacterium nucleatum. The infection was repeated three times at 2 day intervals. Six weeks following the final infection, the maxillary jaws were harvested and alveolar bone loss was quantified using microCT.

Results: The mean of phenotypic response of F2 population (0.0083mm3) was intermediate between the two parental founders, 0.0103mm3 and 0.0062mm3 for A/J and BABLB/cJ, respectively. The results suggest no genomic imprinting (mitochondrial) effect as well as no significant sex effect.

Conclusions: The phenotypic variety of F2 population suggests more than one quantitative trait loci (QTL) affecting host susceptibility to periodontitis. This is the first study which uses mixed infection as experimental periodontitis that: (i) compares different F2 groups (ii) analyze quantitatively a high number of mice by microCT. Currently, we are finalizing the genotyping of 38% of the phenotypic extremes of F2 resource population with 1500 single nucleotide polymorphic (SNP) markers, covering the whole genome which allows subsequently performing QTL mapping analysis. Mapping the QTL will point candidate gene involved in periodontitis.




Sean A. Wiltshire1,2 and Silvia M. Vidal1,2

1Department of Human Genetics, McGill University

2McGill Centre for the Study of Host Resistance, McGill University


In North America, up to 50% of Myocarditis can be attributed to Coxsackie virus B3 (CVB3) infection. Host genetics, viral factors and the environment of their interaction determine variable susceptibility to pathogenesis within exposed human populations. Virally induced pathology in the mouse model mimics the progression of disease in humans. Inbred mouse strains respond differently to CVB3 infection including susceptible strains with A/J background and resistant strains with C57BL/10 background. Through the analysis of a segregating (A/JXB10.A)F2 cross we have recently detected three loci linked to susceptibility to CVB3 induced pathology: Vms1, Vms2 and Vms3 (viral myocarditis susceptibility). We have subsequently confirmed the existence of Vms1 through the use of the consomic substitution strain CSS3 and the localization of Vms1 through the analysis of a segregating (CSS3 x B6)F2 cross. In order to identify causal genes we have undertaken a congenic and sub-congenic approach to dissecting Vms1. Ongoing analysis of potential cell autonomous mechanisms in isolated adult ventricular cardiomycytes may reveal novel immune mechanisms which control infection in the heart.




Association between spatial proximity and functional similarity in human genome

Ekaterina Khraneeva, Andrey Mironov, and Mikhail Gelfand

Faculty of Bioengineering and Bioinformatics, M.V.Lomonosov Moscow State University, Russia

A.A.Kharkevich Institute for Information Transmission Problems, RAS, Moscow, Russia


Recent progress in determination of 3D structure of nuclear chromatin allows one to study correlations between spatial proximity of genome domains and their functional state. We combined the 3D data from (Lieberman-Aiden et al. 2009) with the results of several high-throughput studies of the chromatin functional state. All pairs of regions from different chromosomes were divided into groups according to their proximity, and the distribution of various chromatin marks was calculated within these groups and then compared between the groups.  The results show that, indeed, gene regions that are spatially close tend to have similar patterns of histone modifications (Encode 2004), methylation state (Encode 2004), DNAse hypersensitivity (Encode 2004), and expression level (Lieberman-Aiden et al. 2009). We also analyzed chimeric transcripts as determined by genome mapping of paired-read RNA-Seq data (Xu et al. 2010 and Berger et al. 2010) and observed that the frequency of pairs mapping to two different genome loci is higher among spatially proximal regions. This could be caused by (at least) two reasons: trans-splicing and genome rearrangement compared to the reference genome (either somatic recombination or allelic forms). To test these possibilities we analyzed the data on genome trans-chromosomal rearrangements (Kidd et al. 2008) and demonstrated that again the number of chimeric pairs is higher among spatially close regions, although the effect is weaker than the one observed in the transcriptomic data. Hence, there remains a possibility that the additional chimeras indeed originate from trans-splicing. Finally, we analyzed GO annotations of genes in spatially closest pairs of genome regions. The respective sets were enriched for GO categories related to signal transduction and regulation of transcription. In addition, pairs of spatially close regions had more common GO terms than control random pairs. Taken together, these results seem to demonstrate the presence of co-regulated genome domains formed by regions of different chromosomes.




olfactory receptor gene family evolution in marsupials

Amir Mohammadi, Margaret L. Delbridge, and Jennifer A. Marshal Graves

ARC Centre of Excellence for Kangaroo Genomics and Research School of Biology, The Australian National University, Canberra, Australia


The ability to smell is critical to most mammals, so it is not surprising that the olfactory receptor genes are the largest gene family in genomes of eutherian (placental) mammals. Pouch-bearing mammals (marsupials) diverged from eutherian mammals about 148 million years ago (MYA), so we wished to compare the size and diversity of this gene family between the two mammalian infracalsses, as well as between an American and an Australian marsupial. The complete genomes of two marsupials have now been sequenced, allowing us to look at the conservation of large gene families in this group for the first time. We therefore examined the composition of the olfactory receptor (OR) gene family in the Brazilian short-tailed opossum (Monodelphis domestica) and the tammar wallaby (Macropus eugenii), which diverged about 70 MYA. We isolated all olfactory receptor gene sequences from the first assembly of the Australian model marsupial, the tammar wallaby, classified them by phylogenetic analysis and compared the distribution with the OR repertoire of the opossum. Genomic location of most of ORG clusters was also identified in tammar wallaby by using fluorescence in situ hybridization. Sequence homology, and the genomic distribution of OR genes is well conserved between these two distantly related marsupials. This high level of conservation contrasts with the divergence of ORG families between eutherian mammals with similar divergence times. Our results suggest a significant role for adaptive evolution, rather than genomic drift, in the evolution of chemosensory genes in vertebrates.




Advancing Paternal Age Is Associated with Deficits in Social and Exploratory Behaviours in the Offspring

Rebecca G. Smith1, Rachel L. Kember1, Leonard C. Schalkwyk1, Joseph D Buxbaum2, Abraham Reichenberg1, Cathy Fernandes3*, and Jonathan Mill1*

1Medical Research Council Social Genetic and Developmental Psychiatry Centre, King's College London,    London, United Kingdom

2Department of Psychiatry, Mount Sinai School of Medicine, New York, New York, United States of America

3Department of Psychosis, King's College London, London, United Kingdom

*Contributed equally to this work


Many epidemiological studies have demonstrated an association between advanced paternal age and risk for several psychiatric disorders including autism. In this study we used an animal model to investigate the effects of advanced paternal age on behaviour in the offspring. C57BL/6J offspring were bred from fathers of two different ages, two months old (to represent young fathers) and 10 months old (to represent older fathers) using mothers aged two months in both groups. The offspring underwent open field, holeboard and social recognition tasks to explore differences in their behaviours. We observed that the offspring of older fathers were found to engage in significantly less social and exploratory behaviours than the offspring of younger fathers without any reduction in their motor activity. Brain and peripheral tissues were obtained from both parents and offspring and used to investigate de novo genomic changes associated with increased paternal age using Nimblegen 720K CGH microarrays. Spermatozoa undergo multiple divisions throughout the male lifespan, potentially leading to a higher incidence of de novo copy number variation (CNV) in any resulting offspring. Given recent human data reporting an increased burden of CNVs in autism and other neurodevelopmental disorders, we speculated that such genomic changes could be mediating the behavioural changes we observed. Our study provides the strongest evidence for deleterious effects of advancing paternal age on social and exploratory behaviour and suggests that de-novo chromosomal changes are plausible explanatory factors.



Genome-Wide differential methylation patterns in intersubspecific hybrid mice      

John D. Calaway1,2,3,4, Hyuna Yang5, Elena de la Casa-Esperon7, Megan E. Hanson6, Ezequiel C. Cambranis6, Maria A. Perez-Lamigueiro6, David L. Aylor2, Leonard McMillan8, Gary A. Churchill5, and Fernando Pardo-Manuel de Villena1,2,3,4

1Curriculum in Genetics and Molecular Biology, University of North Carolina  University of North Carolina, Chapel Hill, NC

2Department of Genetics, University of North Carolina, NC, USA

3Lineberger Comprehensive Cancer Center, University of North Carolina, NC, USA

4Carolina Center for Genome Sciences, University of North Carolina, NC, USA

5The Jackson Laboratory, Bar Harbor, ME, USA

6Department of Biology, University of Texas Arlington, Arlington, TX, USA

7Regional Center for Biomedical Research (C.R.I.B.) Albacete Science and Technology Park/ University of Castilla-La Mancha, Spain

8Department of Computer Science, University of North Carolina, NC, USA


Here we report the first genome-wide survey of the mouse differential methylome.  Using the high-density Mouse Diversity genotyping array, we exploited the whole-genome amplification step of the Affymetrix genotyping protocol by adding a pre-digestion step with either methylation-sensitive HpaII or methylation-insensitive MspI endonucleases.  Reported previously as methylation-sensitive single nucleotide polymorphism analysis (MSNP), this technique compares genotyping calls between undigested and HpaII samples to determine allele-specific methylation.  We expanded the analysis by directly analyzing probe intensity data to classify methylation patterns.  By doing so, we identified stably inherited epigenetic states attributed to parent-of-origin or strain-of-origin.  In addition, global methylation analysis suggests that female mice exhibit a higher level of autosomal methylation when compared to males.  This approach identified three novel differential methylated regions (DMR).  We chose one located in the 3Õ UTR of Actn1 for method validation using two independent molecular assays.  Our findings suggest that the mouse Actn1 DMR has moderate tissue and strain specificity and a comparison of the orthologous regions in rat and human reveal a possible rodent-specific DMR.  Lastly, we demonstrate that the Actn1 DMR influences gene expression based on Collaborative Cross mice.





John P. Didion1, John D. Calaway1, Isa Kemal Pakatci2, Leonard McMillan2, and Fernando Pardo-Manuel de Villena1

1Department of Genetics, Lineberger Comprehensive Cancer Center, Carolina Center for Genome Science, University of North Carolina Chapel Hill, NC, USA

2Department of Computer Science, University of North Carolina

Chapel Hill, NC, USA


Cytosine (CpG) methylation is involved in many critical cellular functions, including genomic imprinting, Xchromosome inactivation and silencing of retrotransposons and disease-related genes. Genomic studies increasingly call for consideration of genome-wide methylation patterns, however the methods available for assaying methylation in the mouse are currently limited either by expense or the number and distribution of CpGs interrogated. This later shortcoming is the result of most methylation studies focusing on CpG islands - regions of high CpG concentration whose methylation status tends to be correlated with the activity of nearby genes. However, recent studies have shown that CpGs proximal to islands (as much as 4kb distant) are highly correlated with allele-specific gene expression and thus of great interest to researchers. We have optimized an existing method based on genotyping arrays to assay allele-specific CpG methylation using the Mouse Diversity array and a combination of methylation-sensitive and -insensitive restriction endonucleases. Our method provides fully informative coverage of more than 100,000 CpGs, and partially informative coverage of an additional 90,000 CpGs, most with at least one technical replicate, and roughly even genomic distribution. We present here a general bioinformatic approach for optimization of the Mouse Diversity array to address a range of biological parameters. In addition, we have applied our method to study parent-of-origin and strain-specific effects on methylation in the Collaborative Cross, and have thus generated an optimized data set of untreated controls that we provide as a community resource. Our findings will reduce the expense and challenge of incorporating genome-wide methylation analysis in genomic studies.




Characterisation of Laterality and Cardiac Developmental Defects in the K27 Mutant Mouse

Louise Stephen, K Mitchell, and K Hentges

University of Manchester, UK


Congenital heart defects account for 3% of all infant deaths and many more prenatal. Cardiac development is a highly regulated process, reliant on the cooperation of a vast number of genes. Whilst advances have been made in defining the physiological processes occurring during cardiac development, the genetic regulation is less well understood. Random mutagenesis screens offer a phenotype-driven approach to producing novel mammalian models of human congenital birth defects. We have applied this approach to the study of cardiac development and are currently characterising the K27 mutant mouse, identified from a balancer chromosome chemical mutagenesis screen. The K27 mutant exhibits a recessive embryonic lethal phenotype, apparent from embryonic day (E)9.5, with death occurring by E12. The K27 mutant exhibits a developmental delay and abnormal cardiac morphology. Approximately 50% of mutants demonstrate a reversal of cardiac looping. Analysis of Nkx2-5 and Fgf8 has revealed that both the primary and secondary heart fields are correctly specified in mutants. Likewise K27 mutants express Mef2c in a pattern indistinguishable from control littermates, indicating that cardiac muscle is present in mutants. Laterality defects suggested by the reversal in looping were confirmed by investigation of the expression of genes required for left-right asymmetry, including down-regulation of the Sonic hedgehog-regulated Gli1 gene in mutants. Meiotic mapping has refined the K27 candidate region to a 5 Mb interval on mouse Chromosome 11. This region contains over 100 genes, none of which are currently known to have an established role in cardiac development. Further work will include using high-throughput sequencing to identify the K27 gene.  Continued characterisation of the K27 mutants will define the role the K27 gene plays in cardiac development and left-right asymmetry. Combined these studies will identify a novel gene that contributes to our understanding of cardiac congenital birth defects and of left-right axis formation.



Kyoto Epileptic Rat (KER): A novel rat model of kcna1 mutation showing spontaneous seizures and muscle involuntry contractions

Saeko Ishida1, Tomoji Mashimo1, Takeshi Nishio2, Yu Sakamoto3, Shuji Kaneko 3, and Tadao Serikawa1

1Institute of Laboratory animals,and 2Department of Integrative Brain Science, Graduate School of Medicine, 3 Department of Molecular Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan


Epilepsy is a neurological disorder characterized by unprovoked, recurring seizures with a wide range of manifestations. We have so far performed phenotype-driven ENU mutagenesis in rats to establish animal models of epilepsy. Here we identified a rat exhibiting involuntary muscle movements and convulsive seizures, named Kyoto Epileptic Rat (KER). We characterized KER pathologically and physiologically and identified the responsible gene (Ker) for involuntary muscle movements by positional cloning. KER was backcrossed by more than 10 generations against F344/NSlc background strain to remove latent ENU-induced mutations in other chromosomal regions. In KER, generalized convulsive seizures were observed from 10 weeks of age, and most of the rats died until 7 month of age. By electroencephalogram (EEG) recording, polyspikes and polyspike-and-wave discharges were typically observed during the seizures. By electromyogram (EMG) analysis of the anterior limb muscle, 7 Hz rhythmic spikes were detected during the interictal phase. Using 97 (KER x BN/SsNSlc) x BN backcross progeny, Ker was mapped within the 10Mb region of rat Chromosome 4. Within the region, a missense mutation T925A (S309T) in voltage-gated potassium channel shaker-related subfamily member 1 (Kcna1) gene was identified by sequencing analysis. The mutant homotetrameric Kv1.1 channels were non-functional in Xenopus oocytes. In humans, mutations of KCNA1 cause autosomal dominant episodic ataxia type 1 (EA1) characterized by brief episodes of ataxia associated with continuous myokymia. In addition, the patients with EA1 are ten times more likely to develop epilepsy than normal individuals. In mice, Kcna1 knockout mutation shows a recessive seizure disorders (Smart et al., Neuron 1998), while V408A missense mutation shows no seizures, but exhibits stress-induced loss of motor coordination (Herson et al., Nat Neurosci. 2003). Since KER shows muscular symptoms and spontaneous seizures as an autosomal dominant manner, which are different from the phenotype of the mouse models, KER can be a novel animal model of EA1 to understand the underling mechanism of the KCNA1 diseases.



Identifying modifier genes of mpnsts in the nf1;p53cis mouse model of neurofibromatosis type 1.

Jessica A Van Schaick1,2, Keiko Akagi3, Sandra Burkett1, Christina DiFabio1, Robert Tuskan1, Jessica Walrath1, and Karlyne Reilly1

1National Cancer Institute, Frederick, MD, USA

2The George Washington University, Washington, DC, USA

3The Ohio State University, Columbus, OH, USA


The current study aimed to identify modifier genes of malignant peripheral nerve sheath tumors (MPNSTs) in the Nf1;p53cis (NPcis) mouse model of NF1. Previous studies have shown that the incidence of MPNST development in the NPcis mouse model is affected by the parental transmission of the mutant Chromosome 11. In this study microarray analysis was used to examine gene expression differences between MPNST primary tumors derived from NPcis mice varying in inheritance of the NPcis chromosome from the mother (NPcis maternal) or father (NPcis paternal). Grb10 was found to be more highly expressed in NPcis maternal MPNSTs. Zrsr1 was found to be more highly expressed in NPcis paternal MPNSTs. qPCR was used to validate both gene expression differences. We chose to focus first on Grb10 due to its role as a cytoplasmic signaling adapter protein. Fluorescence in situ hybridization was used to examine the presence of Grb10 on Chromosome 11. Grb10 was found to be lost more frequently in NPcis paternal MPNST cell lines, potentially contributing to the decrease in Grb10 gene expression seen in these tumors. Grb10 is paternally imprinted in the periphery of the mouse, therefore we examined Grb10 isoform expression and found paternal and maternal isoforms expressed in the MPNSTs. Due to these results we are examining whether loss of imprinting is contributing to tumorigenesis. Finally we have generated NPcis;Grb10cis mutant mice. Preliminary studies indicate that NPcis;Grb10cis maternal mice have an increase in PNST incidence and a decrease in survival. Our studies indicate that Grb10 is a candidate modifier gene of MPNSTs, acting in a tumor suppressive manner.





Alexandra Niti1, R Dacquin2, V Rinotas1, P Jurdic2 and Eleni Douni1

1Institute of Immunology, Biomedical Sciences Research Center ŅAlexander FlemingÓ, Vari, Greece

2Institut de GŽnomique Fonctionnelle de Lyon, UMR5242 CNRS/INRA/UCBL/ENS
Ecole Normale SupŽrieure de Lyon, France


Receptor Activator of Nuclear Factor-κB Ligand (RANKL, official symbol TNFSF11) is a central regulator of bone remodelling by mediating osteoclast-induced bone resorption, whereas specific inhibition of RANKL by a fully human monoclonal antibody effectively reduces the incidence of fractures in postmenopausal women. We have recently generated transgenic mice overexpressing human RANKL (TghuRANKL) in order to model human RANKL-mediated pathologies. To achieve a correct pattern of human RANKL expression in the mouse, a transgenic 200kb genomic fragment containing the whole human RANKL gene has been introduced into the mouse genome. Among the five independent transgenic lines obtained, a striking skeletal phenotype was observed in the highest copy number TghuRANKL founder characterized by multiple bone fractures, increased osteoclast numbers, severe trabecular bone loss and cortical porosity. A similar but milder bone phenotype was identified by another high copy TghuRANKL line that developed both trabecular bone loss and cortical bone porosity by the age of 3 months. Additionally, a low copy number TghuRANKL line developed exclusively trabecular bone loss by the age of 3 months. The observed phenotypes in various TghuRANKL lines developed in both sexes, whereas the levels of human RANKL expression were correlated with disease severity. Notably, the osteopetrotic phenotype of mutant mice expressing an inactive form of the endogenous RANKL protein was rescued upon crossing these mice with TghuRANKL mice, showing that the transgenic human RANKL protein is fully active in the mouse. These novel human RANKL transgenic mice represent a unique tool for understanding the pathogenic mechanisms that cause bone resorption and for the evaluation of novel therapeutic approaches targeting RANKL-mediated pathologies such as osteoporosis.



Positive Selection Might Contribute to Functional Divergence of NAIPs Following Gene Duplication in Mammals during the Evolution

Zhaoliang Su1, Chenglin Zhou4, Jianguo Chen2*, Shengjun Wang1*, and Huaxi Xu1*

1Department of Immunology & Laboratory Immunology, Jiangsu University, Zhenjiang, China

2The Affiliated PeopleÕs Hospital of Jiangsu University, Zhenjiang, China

3Zhenjiang Entry-exit Inspection and Quarantine Bureau, Jiangsu, China


The neuronal apoptosis inhibitor proteins (NAIPs) are members of NLRs (Nod-like receptors) protein family. Recent researches have shown that some NAIP genes were strongly associated with both mammalian innate immunity and many diseases, whereas some NAIP genes had been pseudogenization. So the present works were to clarify whether the functional divergence had appeared, as well as whether natural selection played an important role during their evolution. Our results showed that NAIP genes had produced functional divergence and appeared genes duplicated before the divergence; during evolution, lineage-specific duplications of NAIP genes were gradually replaced by pseudogenization and NAIP genes were driven by the positive selection; and the selective force changed the functional sites of NAIP, affected the tertiary structure of NAIP, and then produced the functional divergence. Our results also indicated that the positive selection and functional divergence of NAIP might provide mammals advantages to adapt evolution of geographical environment.




A forward genetics approach identifies A novel member of the DNAJC family that causes neuromuscular disease in mice

Fotios Ioakeimidis, Vagelis Rinotas, Eleni Makrinou, George Kollias, and Eleni Douni

Biomedical Sciences Research Center ŅAlexander FlemingÓ, Vari, Greece


Neuromuscular diseases encompass a wide range of clinical conditions remaining incurable while the genetic and molecular basis of most of these conditions remains unknown. Using mouse ENU mutagenesis we have identified a novel autosomal recessive neuromuscular phenotype. This phenotype manifests with hind limb weakness, progressing with muscle atrophy, reduced body weight, generalized paralysis and death within 30 days after birth. Genetic linkage analysis mapped the causal mutation within a 2Mbp interval containing 24 genes. Sequencing revealed an intronic T>A transversion within a member of the DnaJ homolog, subfamily C (DNAJC) of heat shock proteins (Hsp40s). This is a novel gene with completely unknown function, whose protein is suggested to be located in the mitochondrion, suggestion that is supported by our own confocal imaging data. The intronic mutation generates a novel splicing acceptor site resulting in the insertion of 109 bp in the mature transcript, causing  a frameshift of the last 50 aminoacids of the protein that are replaced by 44 different aminoacids. This DNAJC family member shares 96% identity with its human ortholog, highlighting the functional importance of the protein. Our expression analysis data show constitutive and high expression of the normal gene in all tissues studied, neuronal and non neuronal. To genetically prove the causality of this mutation in our phenotype, we have generated BAC transgenic mice for the human DNAJC gene in order to rescue the neuromuscular phenotype. Our current studies focus on the histological and immunohistochemical characterization of the CNS in our mutant mice, and analysis of the expression pattern and function of the normal and the mutated protein. The identification of a novel DNAJC protein involved in neuromuscular disease and future functional characterization of this protein can shed light into new pathogenetic pathways involved in neuromuscular diseases.






Tyler Buit, Chiao-Ling Lo, Feichen Shen, and Amy C. Lossie

Department of Animal Sciences, Purdue University, West Lafayette, IN, USA


The Odz4 allelic series is comprised of five phenotypically distinct embryonic lethal mouse mutants, which are characterized by abnormal embryonic and extraembryonic mesoderm development, irregular somite formation, imperfect patterning of the skeleton and defective maternal blood flow to the embryo. Odz4 gene regulation is complicated and not well understood. Whole transcriptome and cross-species comparison studies suggest that the 735 kb locus produces over 20 alternatively spliced transcripts, contains over 800 CAGE tags and includes 93 highly conserved non-coding elements (HCEs). These HCEs are >200bp long and exhibit ³70% identity among five mammalian species. Similar highly-conserved DNA sequences can contain mRNA-like non-coding RNAs that control developmental gene expression. We hypothesize that several of these HCEs will encode cis-acting ncRNAs that direct alternative splicing at the Odz4 locus. Our long-term goal is to determine the role of these HCEs in directing Odz4 transcription during development. Using RT-PCR, we demonstrated that 92 of the 93 HCEs are expressed in the adult and/or at various embryonic stages in normal embryos. Each transcript exhibited a unique developmental expression profile. One HCE was detected at the blastocyst stage, which is before the primary Odz4 transcripts are produced. Six HCEs were detected in E6.5 embryos, an important stage in ODZ4 signaling. Northern and RT-PCR analyses indicate that although several of the ncRNAs are new Odz4 exons, at least 60 of these HCEs are expressed sequences of unknown function, all of which lie within 5 kb of an Odz4 CAGE tag. In situ hybridization studies at later time points indicate that these six HCEs demonstrate dynamic expression profiles. For example, at E9.5, Hce39 is restricted to punctate regions in the limbbud, brain and first brachial arch. By E12.5, we detect broad expression across the neural ectoderm, with high-levels at the midbrain-hindbrain junction. RT-PCR and in situ hybridization studies of the HCEs in Odz4 mutant embryos is ongoing, and future studies are aimed at determining the effects of these ncRNAs in specific Odz4 mutants.




Rachel L. Kember, Emma Dempster, Leonard C. Schalkwyk, Jonathan Mill, and Cathy Fernandes

Institute of Psychiatry, Kings College London, UK


Stressful early life environments have consistently been implicated in the development of adverse behavioural phenotypes. Environmental effects can be mediated by epigenetic processes, whereby changes in epigenetic marks due to environment can produce long lasting changes to phenotype. A mouse model is an ideal way to disentangle the contributions of environment, genetics and epigenetics towards phenotypic outcome, as it allows both the genetic and the environmental contribution to be controlled.


The current study investigates the behavioural effects of a model of early life stress (maternal separation) in mouse using two inbred strains (C57BL/6J and DBA/2J). A battery of behavioural tests demonstrates that the maternal separation group exhibit phenotypic differences when compared to controls, including variation in anxiety levels. Corticosterone levels, measured at baseline and post-forced swim test, show increased hormone levels in C57BL/6J maternally separated animals after the forced swim.


We identified three candidate genes that have previously shown associations with early life stress. Epigenetic changes in the glucocorticoid receptor (Nr3c1) and arginine vasopressin (Avp) genes have both been associated with poor maternal behaviour and maternal separation, respectively. Additionally, nerve growth factor IB (Nr4a1) is rapidly induced by various stress responses as an early response mechanism. I will present findings that show significant methylation differences in the hippocampus of maternally separated animals compared to controls at CpG sites in the promoter regions of these genes. Furthermore, preliminary data from a genomewide methylation assay will be presented. Studies such as these will allow further disentanglement of environment, genotype and epigenetic mechanisms in the contribution towards adverse phenotypes. 




Stress mechanisms, inflammation, infection and host resistance

Katia P. Karalis

Biomedical Research Foundation of the Academy of Athens (BRFAA), Greece and ChildrenÕs Hospital, Harvard Medical School, Boston, USA


Stress response, a process well-preserved in mammals, is mediated by the activated hypothalamic-pituitary-adrenal (HPA) axis. Immune system challenges such as inflammatory and infectious stimuli activate stress response leading to release of glucocorticoid. The latter is a very potent immunosuppressive and anti-inflammatory agent when administered exogenously. Glucocorticoid acts through binding to glucocorticoid receptor (GR), two types a and b  have been identified in humans with antagonistic effects in some cases. In addition to the GRs abundance, the local effects of glucocorticoid are regulated by the expression of 11betaHSD, an enzyme that modulates the availability of biologically active levels of glucocorticoid, in a tissue-specific manner. The hypothalamic mediator of the stress response, Corticotropin Releasing Hormone or Factor (CRH/CRF), is also expressed in various peripheral tissues and exerts potent immunomodulatory effects. This factor belongs to a family of peptides acting, although with different affinity, via two G-protein coupled receptors (CRF receptor 1 and 2) expressed in several sites in the nervous system and in a plethora of peripheral tissues. Studies from our and other laboratories have demonstrated the bidirectional, direct and indirect, effects of CRF in inflammatory processes and have started to elucidate the mechanisms mediating these actions of CRF. In this presentation we will review the evidence that supports the critical role of the, so-called, stress hormones in the resistance to inflammatory challenges. Next, we will present our findings from mouse models demonstrating a dual role of the stress system in host resistance and the molecular pathways implicated. Finally, we will discuss the possibilities for novel therapeutic applications for inflammatory conditions, given the challenges provided by the complex regulation of the genes involved in the stress response.



Physiologic consequences of constitutive MTOR inhibition: alterations in cell/organ size and immune cell development

Shuling Zhang1, Julie A. Readinger2, Wendy DuBois1, Mirkka Janka-Junttila3, Richard Robinson1, Magaret Pruitt1, Val Bliskovsky1, Julie Z. Wu2, Kaori Sakakibara1, Jyoti Patel1, Carole A. Parent3, Lino Tessarollo4, Pamela L. Schwartzberg2, and Beverly A. Mock1

1Laboratory of Cancer Biology and Genetics, CCR/NCI, 2Genetic Disease Research Branch, NHGRI, and 3Laboratory of Cellular and Molecular Biology, CCR/NCI, 4Mouse Cancer Genetics Program, CCR/NCI


Mammalian TOR (MTOR) regulates cell growth, proliferation and migration.  Because Mtor knock-outs are embryonic lethal, we generated a hypomorphic mouse with a neo-insertion that partially disrupts MTOR transcription and reduces its expression, thus creating a physiologic model of TORC1/TORC2 inhibition.  Homozygous knock-in mice exhibited reductions in body, organ and cell size.  Although reductions in most organ sizes were proportional to decreased body weight, spleens were disproportionately smaller.  Decreases in the total number of T cells, particularly memory cells, and reduced responses to chemokines suggested alterations in T cell homing/homeostasis.  TCR-stimulated T cells also proliferated less, produced lower cytokine levels, and induced an increased number of regulatory T cells (iTreg).  Surprisingly, B cell developmental effects were more pronounced, with a partial block in B cell development in bone marrow, altered splenic populations, decreased migration to chemokines, and decreased proliferation.  Moreover, AKT phosphorylation was increased in activated B cells, reminiscent of cancers treated with rapamycin.  Furthermore, decreased neutrophil numbers were observed in the spleen, despite normal development and migration in the bone marrow.  Thus, MTOR is required for the maturation and differentiation of multiple immune cell lineages.  These mice provide a novel platform for studying consequences of constitutively reduced TORC1/TORC2 activity.




Tomoji Mashimo, Akiko Takizawa, Birger Voigt, Kazuto Yoshimi, Hiroshi Hiai, Takashi Kuramoto, and Tadao Serikawa

Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Kyoto, Japan


Although the laboratory rat has been widely used as animal models across many fields in biomedical sciences, the inability to utilize germline-competent embryonic stem (ES) cells was a major drawback for research activities that aimed to elucidate gene functions. Recently, Zinc-Finger Nucleases (ZFNs) have been used successfully to create site-specific DNA double-strand breaks, and thereby stimulate targeted gene mutations in a wide variety of organisms including plants, xenopus, drosophila, zebrafish, and rats. Here we demonstrate ZFN-stimulated gene-targeting at an endogenous rat gene, for which human and mouse mutations are known to cause a severe combined immunodeficiency (SCID). Co-injection of mRNAs encoding the custom-designed ZFNs into pronucleus of fertilized oocytes yielded 25% gene-modified offspring, including a wide variety of deletion or insertion mutations. ZFN-modified founders faithfully transmit these genetic changes to the next generation with the SCID phenotypes. The high frequency of gene-targeting and the rapid creation of gene knockouts indicate that ZFN technology can provide a new strategy in rats for creating animal models of human diseases.




A recombination hotspot leads to sequence variability within a novel gene and contributes to complex disease susceptibility

Iris KL Tan, Leanne Mackin1, Nancy Wang1,2, Anthony T Papenfuss3, Colleen M Elso1, Michelle P Ashton, Belinda Phipson2,3, Melanie Bahlo3, Terrence P Speed3, Gordon K Smyth3, Grant Morahan4, and Thomas C Brodnicki1

1St VincentÕs Institute of Medical Research, Fitzroy, Victoria, Australia

2The Walter & Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia

3The Western Australian Institute of Medical Research, Perth, Western Australia, Australia


Type 1 diabetes (T1D) is a multigenic autoimmune disease in which lymphocytes mediate destruction of insulin-producing pancreatic beta cells. Despite the recent success of human genome-wide association studies, genetic heterogeneity and tissue availability still hinder the identification of causative alleles, as well as their effects upon gene function and T1D pathogenesis.  A synergistic approach to ongoing human studies is the use of the nonobese diabetic (NOD) mouse strain, which has provided key insights into T1D.  We have recently positionally cloned a T1D susceptibility locus, termed Idd11, located on mouse Chromosome 4.  Sequence analysis across a critical 6.9 kb interval in a series of congenic NOD mouse strains and in 25 other inbred strains identified several haplotypes, including a unique NOD haplotype, associated with varying levels of T1D susceptibility.  Haplotype diversity within this interval between congenic NOD mouse strains was due to a recombination hotspot that generated four crossover breakpoints, including one with a complex conversion tract.  The Idd11 haplotype and recombination hotspot are located within a predicted gene of unknown function.  This gene, temporarily named Latet, exhibits deficient splicing and decreased expression in relevant immunological tissues of NOD mice.  Notably, it was the recombination hotspot that aided our mapping of Idd11 and confirms that recombination hotspots can create genetic variation representing Ōprivate mutationsÕ and affecting a common polygenic disease.  This finding has implications for studies of complex genetic disease, which may be affected by the ~33,000 estimated recombination hotspots in the human genome.





Randal Westrick, Goujing Zhu, Sara Manning, Angela Yang, David Siemieniak, and David Ginsburg

University of Michigan, Ann Arbor MI USA


Venous thrombosis affects ~300,000 individuals/year in the USA.  A factor V polymorphism, Factor V Leiden, (FVL) is the most common genetic risk factor for venous thrombosis, though it displays only 10% penetrance.  We previously demonstrated synthetic lethality between homozygosity for FVL (FVQ/Q) and heterozygous tissue factor pathway inhibitor deficiency (TFPI+/-) in mice.  To identify modifier genes contributing to FVL penetrance, we used this lethal phenotype for a sensitized ENU mutagenesis screen.  As proof of concept, we tested the loss of one tissue factor (TF+/-) allele to suppress the lethal FVQ/Q TFPI+/- phenotype.  Analysis of 257 offspring from a FVQ/Q X FVQ/+ TFPI+/- TF+/- cross demonstrated that haploinsufficiency for TF suppressed FVQ/Q TFPI+/-.  Thus, suppressor mutations in the TF gene should emerge from our screen.  We next performed a genome-wide mutagenesis screen for suppressors of FVQ/Q TFPI+/-.  Male FVQ/Q mice were exposed to ENU and bred to FVQ/+ TFPI+/- females.  Surviving G1 offspring were analyzed to identify mice with the otherwise lethal FVQ/Q TFPI+/- genotype.  Analysis of 7,128 G1offspring (~2X genome coverage) identified 98 FVQ/Q TFPI+/- mice that survived to weaning.  13 FVQ/Q TFPI+/- G1 mice exhibited successful transmission of a putative suppressor mutation to two or more FVQ/Q TFPI+/- G2 offspring.  11 of these 13 putative suppressors are being subjected to positional cloning to identify the corresponding mutations.  Analysis of 20 offspring from one of the lines resulted in the identification of a Chromosome 3 region encompassing the TF gene (LOD=4.93).  Sequencing of this region is in progress to identify the causative mutation.  Mapping and genome sequencing is underway for the remaining 10 lines.  Identification of these mutations should provide novel insights into hemostatic regulation and suggest candidate modifier genes for Factor V Leiden and other human hemostatic disorders.



Genetic and dietary controls of alternative exon use and mRNA levels in the systems architecture of diet-induced metabolic diseases

Sabrina H. Spiezio1, Annie Hill-Baskin1, Karen Fitch2, Keith Jones2, Michael H. Shapero2, and Joseph H. Nadeau1

1Department of Genetics, Case Western Reserve University, Cleveland, Ohio, USA

2Affymetrix Inc., Santa Clara, CA, USA


Because of their rapidly increasing incidence worldwide, the need is urgent to characterize the genetics and systems biology of obesity and related metabolic conditions such as metabolic syndrome, type 2 diabetes, and non-alcoholic fatty liver disease (NAFLD). Two mouse inbred strains, C57BL/6J (B6) and A/J, show contrasting metabolic responses to high fat vs low fat diets.  Using a complete panel of chromosome substitution strains (CSSs) that we made to study genetically complex traits such as these, we discovered 100s of complex trait genes that have unexpectedly large and non-additive phenotypic effects. Moreover these networks of interacting genes buffer systems properties from genetic perturbations. To examine molecular features of these systems, we compared mRNA and alternative exon profiles in three metabolically relevant tissues, for two diets, and several CSSs and B6 control mice that differ in their response to high fat vs low fat diet. The timepoint was midway through the study period, after 50 days on the test or control diet, but before significant weight gain was evident. Key findings include 103 more mRNAs showing diet-induced differential expression and 104 more diet-induced alternative exon usages in CSSs than B6, cis-effects that were more common than trans-effects on the low fat vs. high fat diet, remarkable heterogeneity in trans-responses to different chromosome substitutions, and finally a modest correlation between changes in mRNA expression and use of alternative exons. Together, these results suggest that changes in alternative exon use are much more common than changes in mRNA levels, that B6 responded in a markedly different molecular manner than CSSs to dietary perturbations, and that alternative exon use may be an important but generally neglected source of functional heterogeneity for modulating molecular and physiological responses to genetic and dietary perturbations.




R Ali1, H Bellchambers1, N Warr2, D Quwailid2, P Denny2 and Ruth Arkell1,2

1Early Mammalian Development, Research School of Biological Sciences, The Australian National University, Canberra, ACT, Australia

2Mammalian Genetics Unit, MRC Harwell, Oxfordshire, UK


The Zic genes are an important family of transcription factors about which little is known. Germ line mutation of Zic genes leads to a variety of congenital defects and somatic mutation of ZICs is implicated in a wide range of cancers. The Zic genes contain a zinc finger domain with supposed DNA binding, protein binding and nuclear localization roles. To learn more about the specific molecular functions of the zinc finger domain we undertook a search for relevant point mutations within the Harwell archive of mutagenised mouse DNAs. The Zic5 gene screen lead to the isolation of 2 missense alleles, one with a conservative amino acid change and one with a non-conservative change. In cell based transcription assays both mutant proteins exhibit a partial loss-of-function and mouse strains harbouring theses mutations have a milder phenotype than a targeted null allele of Zic5. Together this demonstrates that both mutations have generated hypomorphic alleles of Zic5. We have further investigated how a conservative change causes a partial loss-of function. The missense mutation alters a lysine residue and the lysine is a predicted substrate for SUMOylation. In vivo studies indicate that Zic5 is normally SUMOylated at this and other lysines and that this modification is required for localization to the nucleus. Other ZIC proteins have been shown to be trafficked to the nucleus via the importin pathway. Our data therefore point to a previously unrecognized mode of transport for the ZIC proteins. SUMOylation is a post-translational modification that is frequently associated with transcription factors. Previous work has demonstrated that the absence of all SUMOylation is incompatible with development of an organism, however, the effect on individual proteins has been harder to demonstrate. This work provides the first in vivo evidence that SUMOylation is required for the function of an individual protein.




Karen Mitchell, C Clowes, Louise Stephen, and Kathryn E Hentges

Faculty of Life Sciences, University of Manchester, Manchester UK


The coronary vasculature is an essential network providing the blood supply to the heart.  Disruptions in the coronary circulation deprive the working myocardium of oxygen, leading to irreversible damage to cardiac muscle and myocardial infarction.  Cardiovascular disease is a major contributor to mortality worldwide.  The generation of treatments for cardiovascular disease will be aided by a deeper understanding of the developmental processes underlying the formation of the coronary vessels.  We have identified a requirement for Non-Muscle Myosin IIB (NMIIB) in the development of the coronary vasculature.  We isolated an embryonic lethal mouse mutant with Embryonic Hydrocephalus and Cardiac defects (EHC) from a balancer chromosome mutagenesis screen.  Positional cloning revealed that the EHC mutant phenotype is caused by a splice donor mutation in Myh10, which encodes NMIIB. EHC mutants produce an aberrant transcript lacking exon 18, disrupting the reading frame of NMIIB and introducing a premature stop codon.  RT-PCR analysis demonstrates that the mutation destabilises the Myh10 transcript, such that heterozygous embryos preferentially produce wild type Myh10. EHC mutant embryos display cardiac morphological defects such as double-outlet right ventricle.  Additionally mutants do not form coronary vessels, evident from gross inspection of hearts and a lack of vascular endothelial PECAM staining.  The failure to form the coronary vasculature is suggestive of epicardial defects.  Accordingly, EHC mutant epicardial cells fail to form an epithelial layer covering the myocardium, and have disruptions in the localisation of extracellular matrix between the myocardium and epicardium. NMIIB therefore may facilitate coronary vessel formation by supporting adhesive interactions between the epicardium and myocardium mediated via the extracellular matrix.  Future work will determine the mechanisms by which Myh10 deficiency contributes to coronary vascular failure.





Michael J. Parsons, Laura Yates, Jessica Edwards, Lauren Chessum, Charlotte H. Dean, and Patrick M. Nolan

Mammalian Genetics Unit, MRC Harwell, Oxfordshire, United Kingdom


The use of N-ethyl-N-nitrosourea (ENU) mutagenesis screening is a powerful tool for discovering novel genes associated with numerous biological pathways and phenotypes.  The mutant called short circuit (Sci) was found in a dominant enu mutagenesis screen.  This mutant, first characterized in a screen for circadian phenotypes, was found to be homozygous lethal with Sci homozygous animals dying between embryonic day 14.5 to post-natal day one.  The gene containing the Sci mutation encodes for a protein that is a transcription factor with multiple zinc fingers.  This gene has been implicated in various functions including neuronal differentiation and development.  In mouse, this gene is highly expressed in developing brain, lung and heart, with its expression peaking at embryonic day 13.5.  These expression patterns combined with the timing of the lethality in homozygous animals led us to explore the possibility of the Sci mutation interfering with normal lung development, thus leading to the observed homozygous lethality.  Upon conducting a time series of lung development, we found significant decreases in distal airway volume in Sci homozygous animals, as compared to wildtype animals, at embryonic day 18.5 (p<0.05); morphological changes in distal airways were observed as early as embryonic day 15.5. Adult heterozygous Sci animals showed impaired lung function as measured by full body plethysmography following metacholine challenge.  Specifically, heterozygous Sci animals had significant decreases in baseline PenH (a measure of lung resistance), peak inspiratory flow and peak expiratory flow (p<0.05).  We are currently conducting immunohistochemical analysis using lung differentiation and proliferation markers, to further characterize the effects of this gene on lung development.  Taken together these data suggest that this gene may play an important role in lung development.




IDENTIFICATION OF THE DANFORTHÕS SHORT tAIL Mutation using next generation sequencing

Christopher N. Vlangos, Amanda N. Siuniak, Dan Robinson, Arul M. Chinnaiyan, James Cavalcoli, Robert H. Lyons, and Catherine E. Keegan

University of Michigan, Ann Arbor, MI USA


The DanforthÕs short tail (SdSd) mouse first appeared as a semi-dominant spontaneous mutation in an inbred colony at Stanford University in the 1920s.  The phenotype of heterozygous (SdSd/+) animals includes unilateral kidney agenesis, vertebral anomalies, and a shortened and kinked tail.   Homozygous (SdSd/Sd) mice are more severely affected with bilateral renal agenesis, lack of tail, vertebral anomalies, spina bifida, lack of urogenital and anal openings, and persistence of the cloaca.  Homozygous mice are born live but die within 24 hours of birth.  Though genetically mapped to a 1.0cM (1.5Mb) region of mouse Chromosome 2qA3, the Sd mutation has not yet been identified.  Using bioinformatic analysis of the mouse DNA physical map we identified 9 annotated genes with a total of 86 coding exons spanning the corresponding 1.5Mb critical region.  Direct sequencing of the exonic DNA and intron/exon boundaries did not reveal any causative mutations.  Since direct sequencing of the exonic DNA only provided ~1% coverage of the Sd critical region we performed next generation sequencing (NGS) of the entire 1.5Mb region.  Prior to NGS we performed an enrichment capture using an Agilent oligo DNA chip designed to cover all unique (non-repeat masked) DNA of the Sd critical region.  After successful locus specific DNA enrichment, 36bp paired end NGS on an Illumina Genome Analyzer IIx was completed, generating 1.85Gb of sequence.  93% of reads were of proper size and mapped to the enriched region in the proper orientation resulting in an average of 200X coverage.  Analysis of the mapped reads using standard computational techniques did not reveal any causative mutations.  We then turned to interrogation of reads where only a single end of the paired end sequencing mapped correctly to the Sd locus.  By using this novel technique, we were able to identify the presence of an endogenous retroviral-like (ERV) insertion of the MusD family at the Sd locus.  The ERV insertion was confirmed via Southern analysis, and is not present in additional inbred mouse lines tested.  The phenotypic characteristics of homozygous SdSd/Sd mice are comparable to those seen in human patients with caudal regression syndrome, cloacal exstrophy, and VACTERL association.  Thus, the Sd mouse is an excellent model for identification of novel genes responsible for human disorders.




A novel role for Atmin, controlling ciliogenesis through modulation of dynein light chain expression

Paraskevi Goggolidou1, Jonathan Stevens1, Gabrielle Wheway3, Antonella Di Paolo, Rosario Romero1, James Briscoe2, Colin Johnson3, and Dominic P Norris

1Mammalian Genetics Unit, Medical Research Council Harwell, Oxfordshire OX11 0RD, UK

2National Institute for Medical Research, London, UK,

3Section of Ophthalmology and Neurosciences, Wellcome Trust Brenner Building, Leeds Institute of Molecular Medicine, St James's University Hospital, Beckett Street, Leeds, U.K.


Through a phenotype driven genetic screen, we identified the DNA damage response locus Atmin as encoding a zinc finger factor required for normal ciliogenesis. Point mutants destroying Atmin zinc finger structure result in embryonic death around 13.5 days of development. Homozygous mutants exhibit a complex developmental syndrome indicative of a ciliopathy, including exencephaly, coloboma, cardiac outflow tract defects, pulmonary hypoplasia and left-right (L-R) patterning defects. Indeed, impaired ciliogenesis is evident from a decreased number of ciliated cells and shortened cilia in the embryonic node. Notably, these morphologically abnormal cilia retain motility, although L-R patterning is abnormal. Cilial structure is known to be important for SHH signaling and for correct establishment of dorso-ventral (D-V) patterning in the neural tube. A defect in SHH signalling is evident in Atmin mutants, as assessed by GLI3 processing. However, no obvious defects in neural tube D-V patterning were detected, suggesting a specific role of Atmin in GLI3 but not GLI2 processing. We have further identified a dynein light chain as acting downstream of Atmin; mRNA expression analysis shows a ~16-fold downregulation in Atmin mutant embryos. Using si-RNA mediated knock down of Atmin and the dynein, we show that Atmin mediates ciliogenesis through regulation of expression of this specific dynein subunit. Sub-cellular localisation places this dynein subunit in the primary cilium in both cultured cells and the embryo, consistent with it directly influencing retrograde intraflagellar transport.



Genetic Analysis of Complex Traits in the Emerging Collaborative Cross

David L. Aylor1, William Valdar1,#, Wendy Foulds-Mathes1,#, Ryan J. Buus1,#, Ricardo A. Verdugo2,#, Ralph S. Baric3,4, Martin T. Ferris1, Jeffrey A. Frelinger4, Mark Heise1, Matt B. Frieman4, Lisa E. Gralinski3, Timothy A. Bell1, John P. Didion1, Kunjie Hua1, Derrick L. Nehrenberg1, Christine L. Powell1, Jill Steigerwalt5, Yuying Xie1, Samir N.P. Kelada6, Francis S. Collins6, Ivana V. Yang7, David A. Schwartz7, Lisa A. Branstetter8, Elissa J. Chesler2, Darla R. Miller1, Jason Spence1, Eric Yi Liu9, Leonard McMillan9, Abhishek Sarkar9, Jeremy Wang9, Wei Wang9, Qi Zhang9, Karl W. Broman10, Ron Korstanje2, Caroline Durrant11, Richard Mott11, Fuad A. Iraqi12, Daniel Pomp1,*, David Threadgill5,*, Fernando Pardo-Manuel de Villena1,* and Gary A. Churchill2,*

1Department of Genetics, University of North-Carolina-Chapel Hill, Chapel Hill, North Carolina, USA

2The Jackson Laboratory, Bar Harbor, Maine, USA

3Department of Epidemiology, University of North Carolina–Chapel Hill, Chapel Hill, North Carolina, USA

4Department of Microbiology and Immunology, University of North Carolina–Chapel Hill, Chapel Hill, North Carolina, USA

5Department of Genetics, North Carolina State University, Raleigh, North Carolina, USA

6National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, US

7National Jewish Health, Denver, CO, USA

8Oak Ridge National Laboratory, Oak Ridge, TN, USA

9Department of Computer Science, University of North Carolina–Chapel Hill, Chapel Hill, North Carolina, USA

10Department of Biostatistics and Medical Informatics, University of Wisconsin-Madison, Madison, Wisconsin, USA

11Welcome Trust Centre for Human Genetics, Oxford University, Oxford, UK

12Department of Clinical Microbiology and Immunology, Tel Aviv University, Tel Aviv, Israel


The Collaborative Cross (CC) is an eight-way mouse recombinant inbred strain panel that is being developed as a resource for mammalian system genetics.  Here we describe an experiment using partially inbred CC lines to evaluate the genetic structure, mapping power and resolution of the CC. Genome-wide analysis of high density SNP data in the incipient strains reveals high genetic diversity, balanced allele frequencies, and dense, evenly distributed recombination sites – all ideal qualities for a genetic mapping and systems genetics resource.  We mapped white head spotting, a discrete coat color trait; body weight, a highly polygenic complex trait; and more than 11,000 liver gene expression traits.  We demonstrate that analysis based on inferred haplotypes in the eight-way cross population improves power and provides additional information about candidate genes not available from standard crosses or strain surveys.  We were able to identify Kitl as a candidate gene for white head spotting in the WSB/EiJ strain and Asph as a novel candidate gene for body weight.  Genetic mapping of gene expression data demonstrates that resolution in this experiment is on the order of 1Mb, and this will improve in the finished CC panel. The number of eQTL discovered here exceeds all previous efforts at eQTL mapping in mouse strain panels and crosses.  The notable absence of trans- eQTL clustering suggests that multiple functional genetic variants are evenly distributed throughout the genome of CC mice.  We have demonstrated that the unprecedented genetic diversity of the CC, which derives from random mixing of an estimated 45 million SNPs, results in high phenotypic diversity and enhances our ability to map the causative loci underlying complex disease-related traits





realising the potential of mouse disease models – from pathway to therapy

Steve D.M. Brown

MRC Mammalian Genetics Unit, MRC Harwell, Harwell Science and Innovation Campus, OX11 0RD, UK


Classical genetic approaches utilising random mutagenesis coupled with focused phenotyping continues to remain an important tool for discovering novelty in biological systems. Moreover, the development of a comprehensive mutant resource for every gene in the mouse genome, followed by large-scale phenotyping, is also set to transform the available models for dissecting disease states. The discovery of novelty is the key basis for a cycle of discovery comprising genetics, molecular, cellular and physiological studies all of which provide a basis for developing new therapeutic approaches. These paradigms provide a powerful platform for elucidating the genetic pathways underlying disease and bringing new insights to the therapeutic strategies that might be investigated. This is no more important for common diseases for which there is little genetic understanding and for which current treatments are ineffective.


One such disease is recurrent (ROM) and chronic (COME) forms of otitis media. ROM and COME are known to have a strong genetic component, but nothing is known of the underlying genes involved in the human population. Otitis media with effusion (OME) is the most common cause of hearing impairment in children in the Western world, potentially causing language delays, learning and behavioural disruption. The high prevalence of the disease, coupled with its recurrent and chronic nature, accounts for the large number of ventilation tubes inserted into the tympanic membrane of affected children. Placement of ventilation tubes is the most common operation in the UK however the mechanism by which they work remains uncertain.


We have identified two novel dominant mutants, Jeff and Junbo, which develop a conductive deafness due to a chronic suppurative OM. The Jeff mutant carries a mutation in an F-box gene, Fbxo11 (Hum. Mol. Genet. 15, 1-7 2006). Junbo carries a mutation in the Evi1 transcription factor (PLoS Genetics 2: e149 2006). Evi1 represses the TGF-b signalling pathway by the binding of Smad3. Jeff homozygotes die shortly after birth displaying developmental abnormalities including cleft palate, eyes open at birth and impaired lung function. TGF-b signalling is involved in all these epithelial developmental processes and we find that pSmad2 is significantly upregulated in epithelia of Jeff homozygotes. Mice heterozygous for both Jeff and Smad2 mutations recapitulate the Jeff homozygous phenotype. Fbxo11 is known to neddylate p53, a co-factor of pSmad2, and we found that p53 levels are substantially reduced in Jeff suggesting that Fbxo11 plays a role in stabilizing p53. Our findings support a model whereby Fbxo11, via stabilization of p53, is required to limit the accumulation of pSmad2 in the nucleus of epithelial cells.


Hypoxia is a common feature of inflamed microenvironments and the impact of hypoxic mechanisms in OM has received little attention. Responses to hypoxia are mediated via Hypoxia Inducible Factor (HIF) and there is considerable cross-talk between TGF-b and HIF-1a pathways that mediate hypoxic responses. We surmise that dysregulation of TGF-b signalling in the Jeff and Junbo mutations is having an affect on the response of the middle ear, including neutrophil and macrophage function, to the hypoxic environment, leading to the chronic unresolved inflammatory condition. The role of chronic inflammatory hypoxia and hypoxia-inducible factor (HIF) mediated responses were investigated in Junbo and Jeff mouse models. Mice labeled in vivo with Pimonidazole showed cellular hypoxia in middle ear mucosa and inflammatory cells in the middle ear lumen and there was upregulation of Il-1b and Tnf-a that modulate HIF. Hif-1a gene expression was elevated in ear fluid white blood cells and there was upregulation of Vegf pathway genes including Vegfa gene and protein expression. VEGFR signaling inhibitors PTK787/ZK 222584, SU-11248 and BAY 43-9006 reduced hearing loss and modulated inflammatory changes in middle ear mucosa. The effectiveness of VEGFR signaling inhibitors implies HIF mediated VEGF plays a pivotal role in otitis media pathogenesis and targeting molecules in HIF-VEGF signaling pathways has therapeutic potential in the treatment of chronic otitis media. 




importance of Cadm1 and cell adhesion in depressive behavior

C Santos1, B Miller2, M Pletcher3, A Su4, L Tarantino5, and Tim Wiltshire1

1University of North Carolina at Chapel Hill, Division of Pharmacotherapy and Experimental Therapeutics, NC, USA

2The Scripps Research Institute Jupiter, Florida, USA

3Pfizer Incorporated Groton, Connecticut, USA

4Genomics Institute of Novartis Research Foundation at La Jolla, California, USA

5University of North Carolina at Chapel Hill, Department of Psychiatry, NC, USA


Despite evidence for genetic vulnerability to depressive behavior, genes that can predict risk or response to anti-depressants have not been identified. Factors including disease heterogeneity, environmental effects, and complex neurobiological processes complicate studies in mood disorder.  In order to elucidate genetic mechanisms underlying depression, we have performed a series of experiments using a panel of genetically diverse mouse inbred strains. We collected genome-wide gene expression data as well as levels of forty biochemical molecules including neurotransmitters, neurotrophic factors, neuropeptides, and neuroamine regulators and receptors under control, fluoxetine, and treatment-na•ve conditions from brains of inbred mice. Furthermore, we have recorded seven baseline anxiety and depressive behaviors as well as depressive-like responses after administration of fluoxetine or water across 30-35 mouse inbred strains. Since mice within a strain are isogenic, inter-strain genotypic and phenotypic differences are analyzed using haplotype-association mapping algorithm to identify genomic regions associated with the phenotypes of interest known as quantitative trait loci (QTLs). Candidate regions were then prioritized based on convergence of evidence from gene expression, behavior, and biochemical data. A biochemical QTL on Chromosome 9 overlaps with a depressive QTL (which suggests that genes within this region are associated with behavior as well as biological changes associated with differential levels of glutamate decarboxylase 67 (GAD67, official symbol GAD1). There are two genes within this region including cell adhesion molecule 1 (Cadm1) and 2900052N01Rik. Analyses reveal moderate correlation between expression of Cadm1 and anxiety-like and depressive-like behaviors in addition to levels of several biochemical molecules. Furthermore, genes within candidate haplotype-depressive QTL peaks (-logP >3.5) were found to be enriched for genes involved in cell adhesion. Our findings suggest Cadm1 and other cell adhesion molecules may mediate molecular and cellular processes that can result in depressive behavior. Studies are underway to elucidate how cellular adhesive properties modulate biological changes that lead to depression.



An ENU sensitization screen to determine the Physiological function of a- synuclein

Deborah E Cabin, M Casey, and D Zou

McLaughlin Research Institute, Great Falls, MT USA


a-synuclein is a small presynaptic and nuclear neuronal protein that is linked to Parkinson's disease (PD) by its presence in Lewy bodies, the intracellular inclusions characteristic of sporadic PD, and by mutations responsible for rare familial forms of that disease.  The physiological function of a-synuclein is not well understood, though several lines of mice that lack the protein have been studied.  Whether or not a-synuclein's normal function is a factor in PD cannot be assessed until that normal function is better defined.   We have taken a genetic approach to determine a-synuclein's function, a sensitized ENU mutagenesis screen using mice null for the protein.  One hundred twenty pedigrees have been screened for neurological phenodeviants; 45 lines have produced non-sensitized phenotypes, 18 lines that transmit phenotypes are being tested for sensitization, and one confirmed sensitized mutation has been identified.  Among the neurological phenotypes still being tested for sensitization are 2 circling mutants, hyperactivity with odd gait, shaky and poor grip, slow righting and poor grip, lack of exploration, and no response to a clickbox. The confirmed sensitized mutation is in Atp7a  the X-linked trans-Golgi copper transporter mutated in human Menkes disease.  While all males carrying the Atp7a  I610S mutation die regardless of a-synuclein status, Atp7a mutant females that lack a-synuclein die prior to 35 days at a significantly higher rate than Atp7a mutant females that express a-synuclein  (p= 0.00069, C2).  While Atp7a mutations affect many systems in the body, the protective effect of a-synuclein must be neuronal based on its expression pattern.  Menkes disease has a severe neurological component that can progress to decerebration.  In the absence of a-synuclein, brain steady-state levels of both wild type and mutant ATP7A protein are highly variable compared to ATP7A levels in mice that express a-synuclein.  The absence of a-synuclein also appears to cause mislocalization of mutant ATP7A in brain; mislocalization might further impair the function of a hypomorphic protein.  Thus a-synuclein appears to have a neuroprotective function; in Parkinson's disease, sequestration of the protein in Lewy bodies may prevent it from performing that function, which may be particularly important under stress conditions.




Gabor Szalai, Janet Crossland, Jay Coleman, and Michael Felder

Peromyscus Genetic Stock Center, University of South Carolina, Columbia, SC,USA


A Mice of the genus Peromyscus are among the most abundant mammals in North America.  They range from Alaska to Central America and occur in many natural habitats. Though superficially resembling laboratory mice (Mus musculus) and rats (Rattus norvegicus), deer mice are not closely related to either of these species. One of the major advantages of research with Peromyscus is their ready adaptability to colony conditions. Research on this genus has been widespread across so many disciplines that the genus has aptly been referred to as ŅThe Drosophila of North American MammalogyÓ. In order to make Peromyscus as a viable alternative to Mus, we have been working with collaborators on developing comparable genomic resources such as an intermediate resolution genetic map (500 markers to date), synteny map, EST libraries (50 000 EST clones), BAC libraries and  whole genome sequences (to be completed in December, 2010). These resources are used to map the genetic locus controlling the audiogenic seizure sensitivity in deer mice. Epilepsy is a debilitating disease that can arise from either acquired brain lesions or from an inherited susceptibility to cortical hyperexcitability. At least 40-50% of epilepsies have a presumed genetic basis. Although few human epilepsy syndromes are inherited in a simple Mendelian manner, single gene animal models offer valuable opportunities to isolate gene mutations, to identify underlying molecular mechanisms and to explore strategies for therapy. In 1935 a spontaneous recessive mutation appeared among laboratory stocks of Peromyscus maniculatus artemisiae, which has been maintained as a separate stock since. Using homozygousity mapping analyzing a panel 250 F2 and 100 N2 animals we have identified 4 candidate markers. A comparative cytogenetic map between Peromyscus maniculatus and Mus musculus reveal that the M. musculus homologs of the candidate markers on P.m. chromosome 1 and chromosome 23 would be on M.m. chromosomes 7 and 5, respectively. The two mapped mouse audiogenic seizure susceptibility genes, mass1 and jams1 are localized on M. m. chromosomes 13 and 10, respectively. Taken together, our preliminary data suggests that the genetic mutation underlying seizure sensitivity in deer mice resides in a novel gene and is not a homolog of a previously identified susceptibility locus.



mutations in the PDZ domain containing protein gipc3 cause progressive sensorineural degeneration (ahl5 and jams1) in mice and recessive hearing impairment in humans (dfnb95)

Nikoletta Charizopoulou1, Andrea Lelli2, Margit Schraders3,4, Kausik Ray5, Ronald J.C. Admiraal4, Harold R. Neely1, Joseph R. Latoche1, John K. Northup5, Hannie Kremer3,4,6, Jeffrey R. Holt2, and Konrad Noben-Trauth1

1Section on Neurogenetics, 5Section on Signal Transduction, Laboratory of Molecular Biology, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Rockville, MD, USA

2Department of Neuroscience and Otolaryngology, University of Virginia School of Medicine, Charlottesville, Virginia, USA

3Department of Otorhinolaryngology, 6Department of Human Genetics, 4Nijmegen Centre of Molecular Life Sciences and Donders Institute for Brain, Cognition and Behaviour, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands


Progressive sensorineural hearing loss affects the quality of life and communication of millions of people but the underlying molecular mechanisms remain elusive.  Recently, the age-related hearing loss 5 (Ahl5) and juvenile audiogenic monogenic seizure 1 (jams1) loci were shown to underlie progressive hearing loss and audiogenic seizures in Black Swiss mice, respectively.  Here, we identify Ahl5 and jams1 as a 343G>A transition in Gipc3 changing the conserved Gly115 to Arg in its PDZ domain and causing a significant reduction in protein levels.  GIPC3 localizes to inner ear sensory hair cells and spiral ganglia and we show that the mutation disrupts the structure of the stereocilia hair bundle affecting mechano-transduction currents and long-term survival of spiral neurons.  We demonstrate an adverse effect of the Gipc3343A allele on wave I amplitudes of afferent neurons, which we correlate with susceptibility and resistance of audiogenic seizures.  A Gipc3 transgene rescues both hearing loss and audiogenic seizures.  Lastly, we identify a truncating mutation in human GIPC3 in a family segregating autosomal recessive hearing loss DFNB95.  Our study reveals a novel and pivotal role of GIPC3 for the function of the stereocilia hair bundle and for synaptic transmission.



A mutation in the gene encoding mitochondrial Mg2+ channel MRS2 results in demyelination in rats

Takashi Kuramoto1, Mitsuru Kuwamura2, Satoko Tokuda1,2, Takeshi Izawa2, Yoshifumi Nakane1, Kazuhiro Kitada1,3, Masaharu Akao4, Jean-Louis GuŽnet5, and Tadao Serikawa1

1Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University, Japan

2Laboratory of Veterinary Pathology, Osaka Prefecture University, Japan

3Laboratory of Mammalian Genetics, Genome Dynamics Research Center, Graduate School of Science, Hokkaido University, Japan

4Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, Japan

5DŽpartement de Biologie du DŽveloppement, Institut Pasteur, France


The rat demyelination (dmy) mutation serves as a unique model system to investigate the maintenance of myelin, because it provokes severe myelin breakdown in the central nervous system (CNS) after normal postnatal completion of myelination. Here, we identified the dmy mutation and characterized dmy/dmy rats to clarify the pathomechanisms underlying demyelination. By positional cloning, we found that a G-to-A transition, 177 bp downstream of exon 3 of the Mrs2 gene, generated a novel splice acceptor site which resulted in functional inactivation of the mutant allele. Transgenic rescue with wild-type Mrs2-cDNA validated our finding. Mrs2 encodes an essential component of the major Mg2+ influx system in mitochondria of yeast as well as human cells. We found an increased number of mitochondria in the swollen cytoplasm of oligodendrocytes and elevated serum lactic acid concentration in mutant rats, indicating that mitochondria were indeed functionally defective. MRS2-GFP recombinant BAC transgenic rats showed that MRS2 was dominantly expressed in neurons rather than oligodendrocytes, and ultrastructurally observed in the inner membrane of mitochondria. These findings clearly demonstrate that dmy is a loss-of-function mutation of Mrs2 and suggest that dmy/dmy rats suffer from mitochondrial disease. Our findings imply that the maintenance and turnover of myelin are genetically independent from its initial production, and Mg2+ homeostasis in CNS mitochondria is essential for the maintenance of myelin. 




Mark Thomas and Jennifer Harrow

HAVANA Group, Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom


With the publication of the human genome 10 years ago, came the realisation that developmental complexity is not related to gene number. Instead, it would seem that the increased complexity of higher organisms is achieved through greater regulation of more diverse transcripts. Recent studies would suggest that a significant proportion of our genome is transcribed, however only 1-2% of the genome is associated with protein coding loci. A large number of transcripts are therefore non-coding with no known function and are the subject of much debate as to whether they are indeed functional or represent transcriptional noise.

It is the remit of the HAVANA group at the Wellcome Trust Sanger Institute to annotate all transcriptional elements in the human and mouse genomes. As part of this ongoing effort and in collaboration with the Mattick laboratory and HGNC, we are proposing a nomenclature system for long non-coding RNA transcripts (lncRNAs). In the absence of any functional characterisation data for most transcripts, the proposed schema will ascribe a name for each transcript based on its genomic context relative to the nearest protein coding locus. We are also constantly updating our methods to make the most of recent advances in sequencing technology (eg. RNAseq) and structure prediction methods (eg. lincRNAs). This has led to the introduction of new non-coding transcript biotypes, such as antisense, lincRNA, ncRNA-hostgene and 3Õoverlapping ncRNA to further define this rapidly emerging class of genes. Furthermore, analysis of lincRNAs combined with existing data from the Rfam, miRBase and piRNABank databases have identified potentially new hostgenes for these small ncRNAs.



The JAX Cre Repository: Improving the utility of cre driver strains

Stephen A. Murray, Caleb Heffner, Michael Sasner, Cathleen Lutz, Brandon Grossman, Stephen Rockwood, Yashoda Sharma, and Leah Rae Donahue

The Jackson Laboratory, Bar Harbor, ME, USA


Capitalizing on mouse gene targeting projects of the International Knockout Mouse Consortium (IKMC) will require that a large, diverse set of well-characterized Cre driver lines. To fill this need, The Jackson Laboratory (JAX) has committed to increasing the number of Cre lines available to the scientific community. The JAX Cre Repository currently houses and distributes the single largest collection of Cre driver strains totaling more than 200 lines, including 156 that are currently distributed as live colonies. The JAX Cre Repository has embarked on an ambitious project to add value to these strains by comprehensively characterizing Repository Cre lines. Despite the best efforts of those developing new Cre lines, the fidelity of Cre activity is not always ideal. Many difficulties have been reported in various Cre lines, including mosaic or incomplete deletion in a target tissue/cell type, inconsistent activity, expression in non-target tissues, and/or Cre-related toxicity. In many cases, however, this data is not reported or available to the potential user. We have embarked on an ambitious project to add value to these strains by comprehensively characterizing all of the Cre lines distributed. We have developed a comprehensive pipeline for the characterization of Cre driver strains using a LacZ reporter strain in a wide range of tissues and at multiple time points, including both target and non-target tissues. In addition, we are examining possible instances of Cre-induced toxicity, effect of genetic background on excision fidelity, and functional differences that result from maternal versus paternal inheritance of the Cre allele. Thus far we have completed the characterization of 26 strains, and an additional 37 are underway. Our results indicate the vast majority of Cre driver strains exhibit unexpected recombinase activity in a number of tissue types, highlighting the need for extended analysis. We have standardized our data annotation scheme to include 11 broad organ systems, 30 individual organs/structures and 89 substructures, all of which are consistent with the mouse Anatomical Dictionary. Slide-scanned images and associated annotations are published on a dedicated website and submitted to This information will allow users to make informed judgments about the suitability of a particular line for their experiments, and enhance the power of large-scale mouse gene targeting projects.





Petr Danecek, Gan Xiangchao2, Thomas M Keane1, James Stalker1, Binnaz Yalcin2, Martin Goodson2, Sendu Balasubramaniam1, Kim Wong1, Guy Slater1, Andreas Heger2, Eleazar Eskin3, Nick Furlotte3, Chris Ponting, Jonathan Flint2, and David J Adams1

1Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, UK

2Wellcome Trust Centre for Human Genetics, Oxford, UK

3University of California, Los Angeles, USA


The Mouse Genomes Project has sequenced the genomes of 17 inbred mouse strains to between 20-35x coverage, using paired-end Illumina sequencing. One of the major goals of this project is to generate a comprehensive catalogue of all SNPs across these strains. Our strategy for calling SNPs was to employ several next-generation sequencing SNP calling programs and then merge the calls to produce the final callset.  The final set consists of more than 65 millions of SNPs across the strains. We find extremely high concordance with the Perlegen SNP set and using 10Mbp of manually finished sequence for NOD/ShiLtJ find a false positive rate to be 1.4% and false negative rate of 1.7% thus indicating the high quality of our calls. The final call set consists of 24 thousand coding SNPs of which 700 were found to be potentially truncating coding mutations. The wild derived strains such as CAST/EiJ, PWK/PhJ, Spretus/EiJ show an order of magnitude more variation than the classical strains. From the sequenced based SNP calls, we have also created an imputed set of SNPs across a further 94 strains. The full set of SNPs has been uploaded to dbSNP and can be queried on our website which also gives the consequences per SNP, tables of the SNP patterns across the 17 strains, and enables users to visually inspect the raw sequence data surrounding individual SNPs.



Use of PiggyBac-mediated transient transgenic RNAi expression for rapid characterization of gene function during embryonic development

David R. Beier1, Yuko Fujiwara2, Shannon W. Davis3, Haiyan Qiu1, Thomas L. Saunders3, Stuart Orkin2, Sally A. Camper3, and Bryan C. Bjork1

1Genetics Division, Brigham & WomenÕs Hospital, Harvard Medical School, Boston, MA, USA

2Division of Hematology and Oncology, ChildrenÕs Hospital, Harvard Medical School/Howard Hughes Medical Institute, Boston, MA, USA

3Departments of Human Genetics and Internal Medicine, University of Michigan, Ann Arbor, MI, USA


The production of targeted mutations in mice remains the gold standard for the analysis of loss-of-function of specific genes in mammals. However, even with the emergence of large-scale knockout mouse resources, generation of such mutants using embryonic stem cells may still require substantial time and resources. In particular, this approach is difficult to pursue for high throughput applications. For instance, linkage and association studies for mutations or strain-specific traits may yield genomic intervals of only moderate resolution, containing a large number of positional candidate genes. Similarly, microarray analyses typically result in lists of differentially expressed genes, with little indication regarding which ones may be key regulators. An efficient methodology to rapidly screen genes in vivo would enhance the functional analysis of outputs from high throughput screening.  RNA interference (RNAi) is a powerful strategy for studying the phenotypic consequences of reduced gene expression. To develop a method for the rapid characterization of the developmental consequences of gene dysregulation, we tested the use of RNAi for Ņtransient transgenicÓ knockdown of mRNA in mouse embryos. These methods included lentiviral infection as well as transposition using the Sleeping Beauty (SB) and PiggyBac (PB) transposable element systems. Of the three methodologies tested, the PB transposon system produced high numbers of transgenic embryos with the expected phenotype, demonstrating its utility as a screening method. This approach can be useful for phenotypic validation of putative mutant loci, as we demonstrate by confirming that knockdown of Prdm16 phenocopies the ENU-induced cleft palate mutant, csp1. This strategy is attractive as an alternative to gene targeting in embryonic stem cells, as it is simple and yields phenotypic information in a matter of weeks. Furthermore, we found the efficiency of PB for transgenesis was routinely over 65%, suggesting this method has more widespread applications, and we have adapted it for multi-gene knock-down, for expression of affinity-tagged proteins, and for analysis of presumptive transcriptional regulatory sequences.



The Sanger institute mouse genetics programme

Ramiro Ramirez-Solis, J White, E Ryder, R Houghton, and J Bottomley

Mouse Genetics Programme, Wellcome Trust Sanger Institute, Hinxton, UK


The Sanger Institute Mouse Genetics Programme (MGP) capitalizes on the mutant ES cell resources generated by the EUCOMM and KOMP projects by generating and phenotyping mutant mouse lines at a large scale. The program makes the mutant strains and the phenotypic data, available to the scientific community to galvanize deeper analysis aimed at uncovering the molecular mechanisms involved in the phenotypic alterations resulting from the mutant alleles. The MGP has produced over 400 mutant strains, of which more than 200 hundred have finished the phenotypic screen. The phenotypic data can be obtained by visiting the Sanger Mouse Portal ( The website offers the opportunity to download a weekly updated summary heat map that includes all the strains being examined. Scientists are also encouraged to sign up for a phenotypic alert email list to receive early warnings on interesting phenotypes. When two (2) heterozygote mice are genotyped for a particular allele, the strain is advertised to the community at the International Knockout Mouse Consortium (IKMC) website ( Although the specific mice to be distributed may not be available immediately, the MGP seeks to receive notifications of interest, and while the mice are on the shelf, a reasonable effort is made to distribute them to interested parties with minimum delay. Although a majority of strains is currently selected because of prior interest on the gene, we expect that increasingly strains will be selected because of the phenotypic data generated from the MGP. The selection of phenotypic tests included in the program is strongly influenced by an active interaction with experts from the community, and is aimed at exploring a biological space of medical relevance. We will present an update on the program activities and highlight some interesting novel phenotypic findings.



Next level systemic phenotyping of mice

Martin HrabŽ de Angelis and the German Mouse Clinic consortium

Helmholtz Zentrum MŸnchen and Technical University Munich; Institute of Experimental Genetics, Munich, Germany


Next challenges in functional annotation of mammalian genomes are yet of a much larger scope than previous genomics initiatives. Mouse mutant resources must be phenotyped systematically (one after the other) and systemically (assessing all organ systems). In addition, for the next generation of mouse models the ŅenvirotypesÓ, that humans are exposed to need to be modeled. We established the German Mouse Clinic (GMC) as the first mouse phenotyping platform worldwide with the logistics of systemic, standardized phenotypic analysis and interpretation, with open access for the scientific community on a collaborative basis. The German Mouse Clinic is well integrated into EUMODIC and Infrafrontier, pan European projects for functional annotation of mouse models. A new worldwide effort - the International Mouse Phenotyping Consortium – has been established and will move systemic mouse model phenotyping to a new scale. To explore the complex relationship between environmental changes and genetic factors, we have been setting up standardized challenge platforms for mouse phenotyping. By simulating specific environmental exposures or life styles we mimic envirotypes that have a strong impact on human health. Five platforms have been implemented in the areas of nutrition, exercise, air, infection and stress. Goal is to decipher their effects on disease etiology and progression, uncovering the physiological and molecular mechanisms of genome-environment interactions. In addition, identification and non-invasive screening of biomarkers and risk factors for distinct diseases will be a future goal in the GMC using a new metabolomic platform and a new breath-gas analysis set up. First results will be presented.


Gailus-Durner, Fuchs et al. (2005) Nat Methods.

Brown, Chambon, HrabŽ de Angelis and the Eumorphia Consortium; (2005) Nat Genet.

Enard et al. (2009) Cell

Beckers, Wurst and Hrabe de Angelis (2009) Nat. Reviews Genetics




Tooling up MGI for a deluge in mouse phenotype and disease model data

Anna Anagnostopoulos, Janan Eppig, and Mouse Genome Informatics Group

The Jackson Laboratory, Bar Harbor, ME, USA


Global efforts to functionally annotate the mouse genome through large-scale mutagenesis and phenotyping consortia have catapulted the size and diversity of new mouse resources aimed at building predictive models of human disease. The Mouse Genome Informatics resource (MGI, for the genetics, genomics and biology of the laboratory mouse grants free access to current integrated biological knowledge spanning from sequence to phenotype and disease model information to unified data on worldwide mouse resource holdings via IMSR. MGI curates aberrant mouse phenotypes in the context of mutations, strain variations, QTLs and complex traits shaped by intricate epigenome-environment networks. To support the data deluge fueled by ongoing translational research, MGI has devised automated processes to gradually incorporate all multipurpose alleles generated by various IKMC partners, and built a Recombinase Data Portal ( to provide expression and specificity data for each cre-containing transgene and knock-in allele, aiming to integrate phenotype characterization of newly generated mutant mouse lines as available. MGI users can access mouse phenotypes, alleles and disease models of preclinical value using a suite of tools, including an enhanced Quick Search Tool, Mouse GBrowse, web-based vocabulary browsers and data-specific query forms. Robust search inputs include standardized terms from the Mammalian Phenotype Ontology, a widely adopted ontological model that enables phenotype annotations to background-specific allelic genotypes at varying degrees of granularity. Use of human disease (OMIM) terms serves to harness associations between phenodeviant mouse features and orthologous human gene mutations or disease syndromes for which defined mouse genotypes model the human condition. We will review MGI phenotype viewing options, customized retrieval of complex phenogenomic datasets and disease model mining from various perspectives. Supported by NIH grant HG000330




New Approaches to Conditionality

Aris N. Economides, David Frendewey, Peter Yang, David M. Valenzuela, Andrew J. Murphy, and George D. Yancopoulos

Regeneron Pharmaceuticals, Inc. Tarrytown, NY 10591, USA


Conditional mutagenesis is rapidly becoming the method of choice for the study of gene function, but the types of alleles that can be engineered are limited both by vector design and target gene structure.  We therefore developed a new technology — Conditional-by-Inversion (COIN) — that utilizes an optimized invertible gene trap-like cassette, the COIN module.  The COIN module is placed in the antisense strand of the target gene, where it is effectively inert until activated by a recombinase-mediated inversion event that flips the COIN module into the sense strand, thereby disrupting transcription of downstream exons while simultaneously providing a reporter for tracking the mutation.  The COIN module can either be inserted into one of the target geneÕs natural introns (an intronic COIN) or directly into a coding exon as part of an artificial intron (an exonic COIN), greatly increasing allele design flexibility over existing conditional knockout approaches.  The unique artificial intron feature enables the introduction of the COIN module (or other elements) in nearly any location within the target gene, freeing up design choices and increasing the types of alleles that can be designed.  We have constructed COIN alleles for a large number of genes and performed detailed phenotypic analysis on ES cells and mice before and after activation of the COIN modules.  Beyond establishing the robustness, reliability, and broad applicability irrespective of exon-intron structure of this method, our study uncovered rare cases of post-inversion hypomorphic alleles, observed only with intronic COINs, that were caused by ŌskippingÕ of the inverted COIN module, resulting in expression of a mixture of both wild type and COIN-encoding mRNA.  We showed that re-engineering the intronic COIN allele as an exonic COIN by placing the COIN module into the immediate neighboring exon could rectify this problem.  These results not only inform future design decisions, but may explain the hypomorphism occasionally observed with traditional gene trap alleles, which are by nature intronic.  Finally, the exon-splitting and reporter features of COINs open up new engineering modalities for the generation of multifunctional alleles that go beyond conditional-nulls, ranging from simple splitting of exons to introduce novel intronic elements to complex multifunctional alleles with more than two functionalities.  Examples of these novel allele designs will be presented.




Mouse Metabolic Phenotyping Centers:  National Consortium Focusing on Diabetic Complications

Renee LeBoeuf

University of Washington, Seattle, WA, USA


The Mouse Metabolic Phenotyping Center consortium ( is funded by the National Institutes of Health and has the mission to advance research by investigators using mice primarily for diabetes, diabetes complications and metabolic diseases.  There are six science and one bioinformatics centers located across the United States of America which provide unique or well standardized phenotyping tests.  We provide in depth consultations for planning experiments and for data analysis and biological interpretations.  We are also compiling a data base for mouse models studied within our consortium which is open to NIH and industry-contributing researchers.  We support courses including glucose clamping and tracer usage in metabolic research, and have two grant awarding mechanisms.  The Animal Models of Diabetic Complications Consortium, made up of individual investigators who obtain special strains from The Jackson Laboratory, is another group using our services.  Examples of data and MMPC usage will be given. 









Genomic Imprinting: Insights from the GNas CLuster 

Jo Peters

MRC Harwell, Mammalian Genetics Unit, Oxfordshire, UK  


Genomic imprinting results in gene expression according to parental origin. Thus imprinted genes are functionally haploid and so genomic imprinting must confer significant advantage to offset the cost associated with haploidy. Over one hundred and thirty well established imprinted genes are known in the mouse and they tend to occur in clusters.  The Gnas cluster contains two protein coding genes with major phenotypic effects. These are Gnas itself that is maternally expressed in some tissues and Gnasxl that is exclusively paternally expressed. Gnas determines the stimulatory G-protein subunit Gsa and Gnasxl encodes a variant G-protein subunit XLAS. Both GSA and XLAS are signal transduction molecules but they have opposite and potentially antagonistic functions in the regulation of metabolism and behavior, and each has major effects on pre-weaning viability. We used a gene targeting approach to investigate how the imprinted expression of Gnas and Gnasxl is regulated. Imprinted expression within the cluster is under the overall control of an imprinting control region (ICR). This region is DNA-methylated on the maternal allele but unmethylated on the paternal allele. The unmethylated ICR contains an active promoter for a macro noncoding RNA, Nespas, that we have shown is a cis-acting regulator for the paternal allele. This finding adds to other evidence that noncoding RNAs are major elements for regulating imprinted gene expression, although the mechanisms are not well understood. Nespas runs antisense to Nesp, a paternally repressed protein coding transcript and our results show that Nespas is required to silence Nesp so that Gnas can be silenced in turn on the paternal allele. In addition we have shown that Nespas is required for methylation and therefore inactivation of the Nesp promoter. Furthermore, a low level of Nespas can downregulate Nesp in cis through chromatin modification at the Nesp promoter in the absence of DNA methylation.  Our results indicate that a complex set of RNA and epigenetic interactions have evolved to regulate imprinted gene expression.





Anjali Raval, Lisa Korostowski, and Nora Engel

Fels Institute, Temple University School of Medicine, Philadelphia, PA, USA


Long non-coding RNAs (lncRNAs) are a growing category of RNAs with regulatory functions, few of which have been functionally characterized. The Kcnq1 imprinted domain is involved in Beckwith-Wiedemann syndrome. The highly conserved mouse cluster is regulated by a paternally expressed 90kb lncRNA, Kcnq1ot1, that silences most of its neighboring genes, acting strictly in cis. Kcnq1ot1 is transcribed from an intron of Kcnq1 in antisense direction. Intriguingly, the paternal Kcnq1 is released from silencing in a tissue-specific manner, suggesting that strong enhancers can override silencing by the ncRNA. To study the in vivo spatial organization of the domain and provide insight into the complex expression patterns, we carried out chromatin conformation capture (3C) assays. Three-dimensional chromatin topology plays a vital role in bringing wide-spread regulatory elements into close proximity. We anchored the assays at the Kcnq1 and Kcnq1ot1 promoters and tested embryos and neonatal tissues of wild-type mice and mice with a truncation at Kcnq1ot1. Results showed that interaction frequencies are developmentally regulated and exhibit tissue-specificity. In fact, physical contact between the Kcnq1 promoter and specific DNA sequences allowed us to identify novel enhancers. Furthermore, we found major differences in interaction profiles between wild-type and mutant mice. Transcription of the lncRNA physically restricts the regions that the Kcnq1 promoter can interact with in the wild-type mice, whereas mutant mice producing a severely truncated ncRNA exhibit promiscuous contacts and deregulated expression of Kcnq1. These results suggest a unique model whereby the silencing function of Kcnq1ot1 is exerted by restricting the access of promoters to specific enhancers by modifying the flexibility of the chromatin fiber. Nevertheless, strong tissue-specific enhancers can override this effect and establish the three-dimensional contacts required to promote transcription.




AN identity crisis: Abnormal Male germ cell Development and the initiation of testicular cancer

Jason Heaney, Jean Kawasoe, Megan Michelson, and Joseph Nadeau

Case Western Reserve University, Department of Genetics, OH, USA


Testicular germ cell tumors (TGCTs) are the most common cancer in young men.  TGCTs result from anomalies in the development of primordial germ cells, totipotent embryonic stem cells that normally differentiate into mature gametes.  Spontaneous TGCTs occur at an appreciable frequency only in the 129 family of inbred strains.  TGCTs in mice initiate around embryonic day 13.5 (E13.5) during the same developmental time-point at which germ cells commit to meiosis or mitotic arrest (the mitotic:meiotic switch).  In females, embryonic oocytes pluripotent gene expression decreases and Stra8 expression is induced to initiates entry into meiosis.  In males, pluripotent gene expression decreases, Stra8 is not induced and gonocytes, the embryonic precursors of adult male germ cells, become quiescent until after birth when they differentiate to form spermatogonia.  Given the overlap in the timing of TGCT initiation and the decision to enter meiosis or mitotic arrest, we tested whether a defect in the mitotic:meiotic switch contributes to TGCT development.  We discovered that TGCT susceptible gonocytes prematurely express markers of adult spermatogonia and embryonic oocytes between embryonic days E13.5 and E16.5.  However, the similarities in differentiation stop at a stage resembling Ngn3-positive, Kit-negative, undifferentiated A spermatogonia.  A sub-population of TGCT susceptible gonocytes express Ccnd2 but not Kit.  Because expression of both genes occurs at and contributes to male germ cell differentiation to meiotically-commitment, differentiated A spermatogonia, the ultimate fate of TGCT susceptible gonocytes may be altered from meiotic to tumorigenic.  To test whether prematurely differentiating, TGCT susceptible gonocytes commit to meiosis, we immunolabeled chromosome spreads for components of the synaptonemal complex (SCP3 and SCP1). Interestingly, axial elements form but synapsis does not occur in TGCT susceptible gonocytes.  These results suggest that prophase I of meiosis is initiated but aborted in TGCT susceptible gonocytes. We recently discovered that suppression of pluripotent gene expression is delayed in TGCT susceptible gonocytes.  Interestingly, in vitro stimulation of pluripotent gene expression in undifferentiated A spermatogonia induces the formation of pluripotent stem cells with teratoma forming capacity.  Similarly, TGCT formation in vivo may result from the establishment of A spermatogonia-like cells with pluripotent stem cell properties.  Ongoing studies are testing the influence of retinoic acid on TGCT susceptibility and the contributions of Stra8 expression and meiotic initiation to TGCT formation.




Toyoyuki Takada1, Akihiko Mita1, Shigeharu Wakana2, Kazuo Moriwaki2, Hiromichi Yonekawa3, and Toshihiko Shiroishi1

1National Institute of Genetics, Shizuoka, Japan

2RIKEN BRC, Ibaraki, Japan

3Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan 


Energy metabolism-related traits including fat deposition are governed by environmental and complex genetic factors, which are also largely influenced by aging with concomitant change in basal metabolic rate. In this study, we intend to uncover the genetic determinants that control the age-associated changes of energy metabolism by functional genomics with mouse inter-subspecific consomic strains. We have established a full set of consomic (chromosome substitution) strains, B6-ChrNMSM, with each of C57BL/6 (B6) chromosomes replaced by its counterpart of MSM/Ms, which was derived from M. m. molossinus. Using the panel of consomic strains, we conducted systematic phenotype screening of body growth and physiological traits by 10 weeks of age, and successfully detected several hundred of energy metabolism-related QTLs ( Currently, we extend the period of observation of the same traits to 25 weeks of age. Comparing phenotype of each strain at the two different observation periods, we systematically explored strains that show significant change in measurement values. We found that many strains drastically changed the values of energy metabolism-related traits when compared with change of the consomic background strain B6. For examples, consomic strains of chromosome 11 and 15 showed extremely low rate in increase of fat deposition. This suggests that although wild-derived MSM/Ms strain has thrifty-type metabolic pathway as a whole, it also carries genetic factors leading to age-associated anti-obesity. Thus, this study shows that the inter-subspecific consomic strains would provide powerful tools to dissect age-associated change of energy metabolic-related traits.




Rong Yuan1, SW Tsaih1, Q Meng1, K Flurkey1, J Nautiya2, SB Petkova1, MA Bogue1, KD Mills1, LL Peters1, CJ Bult1, CJ Rosen1, JP Sundberg1, M Parker2, DE Harrison1, GA Churchill1, and B Paigen1

1The Jackson Laboratory, Maine, USA

2Imperial College London, London, UK


To better characterize aging in mice, the Jackson Aging Center carried out a lifespan study of 31 inbred strains. Clinical assessments were carried out every 6 months, measuring multiple age-related phenotypes including development traits, organ functions, body composition, hematology, hormonal levels, and immune system parameters. In a concurrent cross-sectional study of the same 31 strains at 6, 12, and 20 months, more invasive measurements were carried out followed by necropsy to assess apoptosis, DNA repair, chromosome fragility, and histopathology. Survival curves varied dramatically among strains. The median lifespans ranged from 251 to 964 days. By conducting correlation analyses, we found some phenotypes correlated with lifespan. Particularly, plasma IGF1 levels showed an inverse correlation with a median lifespan at 6 months (R =-0.33, P = 0.01). This correlation became stronger if the short-lived strains with a median lifespan < 600 days were removed (R = 0.53, P < 0.01). IGF1 levels at 6 months also significantly correlated with the ages of vaginal patency (AVP), a measurement for sexual maturation (R=-0.44, P=0.01). AVP positively correlated with median lifespan, however it is not significant (P=0.22) until the short-lived strains were excluded (R=0.38, P<0.05). Haplotype association mapping identified three loci — Vpq1, 2, 3 — on Chromosomes 4 and 16 that significantly associated with the variation in AVP. At these loci, all four wild-derived strains — WSB/EiJ, CAST/EiJ, PWD/PhJ, MOLF/EiJ — each of which represents a different sub-species of the mouse family, have the same haplotypes, which differ from those of almost all domesticated inbred strains. A consomic strain, which carries chromosome 16 of PWD/PhJ on the C57BL/6J background, had significantly delayed AVP. Bioinformatic analysis suggested that nuclear-receptor-interacting protein 1 (Nrip1) is a candidate gene for Vpq3. AVP was delayed in the Nrip1 knock out mice compared to controls, demonstrating that Nrip1 can influence the age of sexual maturation. These results suggest common genetic mechanisms may exist for regulating IGF1 levels, AVP and lifespan. Using aging related phenotypes as entrŽes may facilitate the identification of genes that regulate aging and longevity.




Subspecific origin and haplotype diversity in the laboratory mouse

Hyuna Yang1, Jeremy R Wang2, John P Didion3, Ryan J Buus3, Timothy A Bell3, Catherine E Welsh2, Franois Bonhomme4, Alex Hon-Tsen Yu5, Michael W Nachman6, Jaroslav Pialek7, Priscilla Tucker8, Pierre Boursot4, Leonard McMillan2, Gary A Churchill1, and Fernando Pardo-Manuel de Villena3

1The Jackson Laboratory, Bar Harbor, ME 2Department of Computer Science; 3, Department of Genetics, Lineberger Comprehensive Cancer Center, Carolina Center for Genome Science, University of North Carolina Chapel Hill, NC, USA

4UniversitŽ Montpellier, Institut des Sciences de l'Evolution, Montpellier, France

5Institute of Zoology and Department of Life Science, National Taiwan University, Taipei Taiwan ROC 10617

6Department of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ, USA

7Department of Population Biology, Institute of Vertebrate Biology, Studenec, Czech Republic

8Department of Ecology and Evolutionary Biology, University of Michigan, Ann Arbor, MI, USA


In this study we provide the first genome-wide, high-resolution map of the phylogenetic origin of the genome of the majority of extant laboratory mouse inbred strains. Our analysis is based on the genotypes of wild caught mice from three distinct subspecies of Mus musculus.  We demonstrate tha classical laboratory strains are derived from a small pool of fancy mice with limited haplotype diversity.  Their genomes are overwhelmingly M. m. domesticus in origin and the remainder is mostly of Japanese (M. m. molossinus) origin.  We have generated genome-wide maps of haplotype sharing in classical inbred strains based on identity by descent from fancy mice and demonstrate that, despite broad phenoptypic diversity, classical inbred strains have limited and non-randomly distributed genetic diversity. In contrast, wild-derived laboratory strains represent a broad sampling of diversity within the species M. musculus.  However, intersubspecific introgression is pervasive in these strains and contamination by laboratory stocks has played an important role in this process. The subspecific origin, haplotype diversity and identity by descent map in laboratory strains can be visualized and searched at  This work provides a detailed characterization of the evolutionary origins of this widely used model for biomedical research.





Genetic architecture of hybrid sterility: MATCHING the puzzle pieces together

Jiri Forejt, Maria Dzur-Gejdosova, Vaclav John, Tanmoy Bhattacharyya, Petr Simecek, Sona Gregorova, Ondrej Mihola, Petr Flachs, and Zdenek Trachtulec

Institute of Molecular Genetics and Center for Applied Genomics, Academy of Sciences of the Czech Republic,Videnska, Czech Republic


Hybrid sterility genes are involved in reproductive isolation and consequently in speciation by restricting gene flow between related taxa. The house mouse Mus m. domesticus and Mus m. musculus, two subspecies in statu nascendi, represent unique mammalian models to study speciation because of their recent evolutionary divergence, and because of the availability of variety of genetic tools and sequence information. We are using C57BL/6J (B6) inbred strain as a model of Mus m. domesticus (cca 95% of its genome is of domesticus origin) and PWD/Ph wild-derived inbred strain of musculus origin. All F1 hybrid males from crosses of PWD females and B6 males are sterile with no sperm. By substituting B6.PWD-Chr# consomics for B6 parent in PWD x B6 hybrid cross, we tested, separately for each chromosome, the necessity of PWD/B6 heterozygosity for male sterility. Using these crosses we found that PWD/PWD homozygosity of Chr 17 or Chr 19, and B6 form of Chr X rescue hybrid sterility while PWD/PWD genotype of all other autosomes did not interfere with sterility of hybrid males. In independent experiments, we mapped hybrid sterility genes as QTLs using several genetic backcrosses, F2 intercrosses and testcrosses. At least 3 hybrid sterility loci were identified, in agreement with the consomic mapping. On Chr 17 we identified Hybrid sterility 1 locus with Prdm9 gene and, more recently, we localized the Hstx2 gene on Chr X. Genetic mapping of the gene(s) responsible for hybrid sterility rescue on Chr 19 is in progress. Detailed analysis of intrameiotic block characteristic for (PWD x B6) hybrid male sterility revealed failure of meiotic pairing of variable number of autosomes. The unpaired autosomes, differentially decorated by SYCP1 and SYCP3 antibody, displayed failure of DSB repair at pachynema marked by the persistence of ATR, RAD51 and gH2AX. The X chromosome showed lack of di-methylation of histone H3K9, and transcription profiling of prepubertal testes and FACS- sorted primary spermatocytes indicated improper inactivation of the X chromosome. Thus the interference with meiotic male sex chromosome inactivation (MSCI) could represent the major surveillance mechanism of hybrid sterility that could be under the control of at least three hybrid sterility genes. We want to propose that the interference with the MSCI is the major molecular mechanism explaining the HaldaneÕs rule in gametogenesis of interspecific hybrids.



molecular interactions of dead end (DND1)

Angabin Matin1, Zhu Rui1, KangLi Luo1, Chitralekha Bhattacharya1, Michelina Iacovino2, Elisabeth Mahen2, Michael Kyba2 and Sita Aggarwal3

1University of Texas, MD Anderson Cancer Center, Houston, Texas, USA, 2University of Minnesota, Minneapolis, USA

3Pennington Biomedical Research Center, Baton Rouge, Louisiana, USA


Inactivation of the Dead End (Dnd1) gene in mice results in sterility or development of testicular germ cell tumors in a strain specific manner.  Dnd1 encoded protein, DND1, possesses canonical RNA recognition motifs.  It has been shown that DND1 binds to the 3Õ-untranslated regions (UTRs) of mRNAs to block microRNA (miRNA) mediated inhibition of translation.  As DND1 helps maintain translation from specific mRNAs, it is expected that inactivation of DND1 will result in loss of cell-type specific protein expression and cell death. DND1 is expressed in germ cells as well as in embryonic stem (ES) cells. Early germ cells and ES cells share gene expression patterns, markers and miRNAs in common. For example, both cell types express markers such as POU5F1, VASA, NANOG and FRAGILIS.  They express similar miRNA families (example, miR 209-295, miR 302-367 and miR 17-92) and pluripotency factors such as POU5F1, SOX2 and NANOG.  Indeed, there is speculation that ES cells may be derived from germ cells of the early embryo. Because ES cells express DND1, it is expected that they also express the mRNAs and miRNAs whose activity is modulated by DND1.  Therefore, we generated a stable, modified ES cell line that expresses tagged DND1 in a regulatable manner.  We used this line to perform ribonucleo immunoprecipitation (RIP) assays followed by RT-PCR.   We found that the mRNA targets of DND1 in ES cells include pluripotency factors, apoptotic factors and tumor suppressors.  We will present our RIP results of candidate DND1 mRNA targets from ES cells. Because protein levels of many factors are precisely regulated in ES cells, our results suggest that DND1 likely imposes another level of translational regulation of critical factors in ES cells.    Second, we examined the effect of DND1 interaction with the cytidine deaminase, APOBEC3.  We had previously demonstrated that APOBEC3 interacts with DND1.  Using luciferase reporter assays we found that APOBEC3 can block DND1 activity and restore miRNA mediated translation repression of p27.  Based on our results we hypothesize that the APOBEC3-DND1 interaction may be involved in providing specificity of DND1 function.  In summary, our results indicate that DND1 likely imposes another level of translational regulation to control expression of critical factors in ES cells. In addition, interaction of DND1 with other proteins such as APOBEC3 may provide further specificity of DND1 function. These studies are beginning to shed light on the role of DND1 in cell death and transformation. 



Genetic dissection of Metatasis susceptibility converges on a common mechanism

J Alsarraj, S Winter, N Goldberger, K Mattaini, M William, L Lukes, R Walker, and Kent W Hunter

Laboratory of Cancer Biology and Genetics, National Cancer Institute, NIH, Bethesda, MD, USA


Metastasis, or dissemination and growth of tumors at secondary sites, remains a major unsolved problem that accounts for the majority of cancer related mortality.  The metastatic process is a complex cascade of events that includes escape from the primary tumor, invasion through surrounding tissue, penetration of the vasculature, resistance to anchor-independence induced apoptosis, arrest in the secondary site, exiting from the vasculature and finally growth in a novel microenvironment.  Acquisition of these abilities has traditionally been thought to be due to the accumulation of somatic mutations in a small subset of cells during tumor evolution.  More recently however, our laboratory has demonstrated that there is a significant inherited susceptibility for breast cancer metastasis in both mouse and human populations.  Using a quantitative trait strategy we have identified and validated a number of metastasis susceptibility genes, including the genes Sipa1, Brd4, and Rrp1b.  Applications of additional genome-wide analytical tools have revealed additional candidate metastasis susceptibility genes from our genetic mapping crosses.  Unexpectedly, many of the genes identified appear to encode proteins that operate within the same transcriptional control and chromatin modification pathway, physically interact, and oppose each others function.  These data suggest that inherited metastatic susceptibility is due to modification of a complex mechanism regulating basal transcriptional control of breast epithelial cells. 



Icst is a dominant negative mutation of lmx1b

Sally H. Cross1, Lisa Mckie1, Margaret Keighren1, Dan Macalinao2, Alison L. Kearney2, Simon W. John2,3, and Ian J. Jackson1

1MRC Human Genetics Unit, Edinburgh, UK

2The Jackson Laboratory, Bar Harbor, ME USA

3Howard Hughes Medical Institute, The Jackson Laboratory, Bar Harbor, ME USA


The ENU-induced mutation iris-corneal strands (Icst) causes raised intraocular pressure, bulging eyes and anterior segment scarring when heterozygous and is recessive lethal. Homozygotes have limb and skull abnormalities and absent cerebellum. Icst is a missense mutation of the LIM-homeodomain transcription factor Lmx1b that prevents the binding of the mutant protein to its target DNA sequence. Mutations in LMX1B underlie the dominant human disorder nail-patella syndrome (NPS). This is a pleiotrophic disorder characterised by nail and joint dysplasia that is sometimes accompanied by kidney defects and glaucoma. It is thought to be caused by haploinsufficiency, although there is great variability in disease aetiology both within and between families. Lmx1b has been knocked-out in the mouse and although the reported homozygous phenotype is very similar to that of Icst, heterozygous knock-out mice are reported to be normal, in contrast to the strongly penetrant eye phenotype of Icst heterozygotes. Both mutant models are on C57BL/6 suggesting strain background is unlikely to account for the observed phenotypic differences. We hypothesised that Icst mutant LMX1B protein has a dominant negative mode of action and that the relative level of wild-type to mutant protein is crucial. We used recombineering to introduce the Icst mutation into an Lmx1b-containing BAC and made transgenic lines with both the wild-type and the mutant BAC. Both express the Lmx1b transgene at 65-70% the level of the endogenous gene. One copy of the wild-type BAC rescues the dominant eye phenotype and two copies rescue the homozygous lethality but not the limb phenotype. The limbs of transgenic rescued homozygotes are symmetrically ventralised, indicating that although the absolute level of wild-type LMX1B protein is greater than in Icst heterozygotes, the excess mutant protein prevents normal development. Furthermore, when we introduce the mutant Lmx1b transgene into Icst heterozygotes we see embryonic lethality, demonstrating that a normally sufficient, heterozygous, wild-type gene is insufficient when there is an increased dose of mutant protein. It has previously been suggested in humans that mutant LMX1B alleles do not influence the wild-type allele. However, we show that the gene can have dominant negative mutant forms, and some cases of NPS could be due this mechanism.




elucidating the role of nsdhl and cholesterol metabolism in cns development using a conditional knOckout allele

Gail E Herman1, N Bir1, L Binkley1, K McLarren2, C Boerkoel2, and D Cunningham1

1The Research Institute at Nationwide ChildrenÕs Hospital, Columbus, OH, USA

2Department of Medical Genetics, University of British Columbia, Vancouver, BC, USA


NSDHL (NADH steroid dehydrogenase-like) is a 3β-sterol dehydrogenase involved in the removal of C-4 methyl groups in one of the later steps of cholesterol biosynthesis. Mutations in the murine gene are associated with the X-linked, male-lethal mouse mutations bare patches (Bpa) and striated (Str). Intellectual developmental disabilities (IDD) and developmental CNS malformations are prominent features of human disorders of cholesterol synthesis. In addition, abnormalities of cholesterol metabolism have been implicated in more common CNS disorders ranging from autism to AlzheimerÕs. Recently, two families with X-linked IDD in males have been found to have hypomorphic human NSDHL mutations. In one family, female carriers exhibited significant callousness (P=0.002), linking cholesterol and behavior. To begin to understand the role of cholesterol and its intermediates in CNS development and behavior, we generated a conditional Nsdhl targeted allele (Nsdhlflx5). The ŅfloxedÓ animals are fertile and without any obvious phenotype. Nsdhlflx5/+ females mated to Sox2-Cre males exhibit an identical phenotype to that observed in surviving Bpa1H/+ females carrying a null allele. There was also no detectable NSDHL protein in Sox2-cre deleted E9.5 or E10.5 male embryos. Nsdhlflx /Nsdhlflx5 females were mated to nestin, GFAP, or Thy1 cre males that have cre expression in neural precursors starting at E10.5; radial glia at E13.5; and selected cortical neurons at P15, respectively. No liveborn deleted NsdhlnesΔ5/Y have been recovered, while Thy1 cre deleted mice have no overt phenotypes at >100 days of age. Hemizygous GFAP cre deleted males develop early postnatal ataxia (P10-12), wasting, and cerebellar degeneration with massive apoptosis and death by P15-P20. While early cerebellar differentiation appears normal, cells of the external granule layer do not appear to exit the cell cycle and migrate properly. In addition, although GFAP cre is not expressed in Purkinje cells, these neurons also die, suggesting that they require the continued presence of supporting glia or are exposed to toxic metabolites. The hippocampus similarly exhibits neuronal cell loss beginning in the late prenatal period. Possible mechanisms, including altered cell signaling and accumulation of toxic intermediates, will be discussed. 



Genetic, stem cell, and systems analyses of neurodegenerative diseases

George A Carlson1, R Bennett1, ME Orr1, I Lee2, H Yoo2, J-H Cho2, D Hwang3, and LE Hood2

1McLaughlin Research Institute, Great Falls, Montana, USA

2Institute for Systems Biology, Seattle, Washington, USA

3POSTECH, Pohang, Republic of Korea


Prions cause transmissible neurodegenerative diseases and replicate by conformational conversion of benign forms of prion protein (PrPC) to disease-causing PrPSc isoforms. Tracking global gene expression changes in the brains of 8 distinct mouse strain-prion strain combinations that differed widely in disease incubation time enabled capture of the effects of prion strain, host genetics, and PrP concentration. A core of 333 differentially expressed genes (DEGs) appear central to prion pathogenesis.  Most of these core DEGs appear before clinical disease but are related to pathological processes.  Core DEGs also change in PrP-overexpressing transgenic mice with very short incubation times, but the magnitude of differential expression is smaller and closer in time to clinical illness than in other mouse strain-prion strain combinations. The most significant DEGs in these transgenic mice change early and are not seen in the other combinations; we hypothesize that these DEGs may reflect the earliest responses to prion infection and that their gene products may be involved in prion replication.  To test this, CNS stem cell-containing neurosphere cultures were infected with mouse prions. Prior to induction of differentiation, no obvious pathogenic effects of prion replication on neurospheres were observed; thus, DEGs found in prion-infected neurospheres likely reflect processes related to prion replication.  This systems approach is enabling identification of intersecting networks of genes crucial for establishing and maintaining prion infection and will help define the link between replication and pathogenesis.  Similar approaches are now being applied to a mouse model for frontotemporal dementia with the ultimate goal of identifying gene and protein expression signatures that can discriminate among diverse neurodegenerative diseases.





Deanna Acosta1, Melissa J Fazzari1, John M Greally1, and Cristina Montagna1

1Albert Einstein College of Medicine, Bronx, NY, USA


Postnatal development of the mammary gland is a complex and highly specialized process characterized by cell proliferation and tissue remodeling. Many genes involved in mammary gland development have been identified and their expression extensively investigated at all major developmental stages.  However, the means by which these genes are regulated have not yet been established. Despite advances in understanding the role of epigenetics in gene regulation, little is known about the relationship between cell subtype-specific cytosine methylation and gene expression in the developing mammary gland. Based on previous reports that cytosine methylation is dynamic during development and that cell type-specific methylation affects gene expression, we hypothesize that alterations of DNA methylation levels occur during postnatal mammary gland development in a stage- and cell subtype-specific manner. We propose that these changes act as one of the mechanisms regulating the expression of transcription factors that drive development and differentiation. Using a known gene in breast development (Serpinb5) we determined if cytosine methylation changes occur during mouse mammary gland development and if these changes correlate with expression. Serpinb5 is regulated by alterations in cytosine methylation in breast cancer cells. Our preliminary studies suggest that the Serpinb5 promoter region is differentially methylated through the developmental stages and that its methylation status correlates with changes in expression. To identify novel candidate loci, we performed a genome-wide analysis of cytosine methylation levels during mammary gland development. Our study indicates that conserved, non-coding regions around the transcription start sites of a subset of genes undergo changes in methylation during development.  Of these, we selected Serpinb5, Gata3, Tnfsf11 (Rankl) Cebpb, and Cebpd because they have been shown to play critical roles in mammary gland development whereas Gata2, a novel candidate gene, is involved in the development of other cell types. To approach the issue of mammary gland tissue heterogeneity, we sorted luminal and myoepithelial cells from different developmental stages. With these cell subtypes, we aim to establish a genome-wide methylation profile and validate the role of cytosine methylation in the regulation of our candidate genes during mammary gland development. This study would not only uncover the role of cell subtype-specific cytosine methylation in tissue morphogenesis, but it will help pinpoint genes that could be altered at early stages of breast tumorigenesis.



Toward an unified measure of intraspecific selective pressure

Roberto Amato1,2, Gennaro Miele1,2, Michele Pinelli1,3, and Sergio Cocozza1,3

1Gruppo Interdipartimentale di Bioinformatica e Biologia Computazionale, Universitˆ di Napoli "Federico II" & Universitˆ di Salerno, Italy

2Dipartimento di Scienze Fisiche, Universitˆ di Napoli "Federico II" - INFN Sezione di Napoli, Naples, Italy

3Dipartimento di Biologia e Patologia Cellulare e Molecolare "L. Califano", Universitˆ di Napoli "Federico II", Naples, Italy


During recent decades the study of human evolution has been of increasing interest, also due to the large amount of data now available. In the mean time, new applications of evolutionary biology to medical problems are being discovered at an accelerating rate. Several estimators for the selective pressure have been proposed. Being introduced to face different aspects of selective pressure, each measure has its own pros and cons. We focused on the most widely used intraspecific estimators of selective pressure. In particular, to cope with both inter- and intra-group phenomena, we analyzed the fixation index (FST) and some measures based on the extended haplotype homozygosity (namely REHH, iHS, XP-EHH). We assessed the performances of each measure on simulated data produced using Fregene, a tool developed and calibrated to reproduce, in a biologically sound manner, our evolutionary history, also taking into account for complex demographic, selection and recombination scenarios. By using GRID facilities to produce a large enough amount of data, we were able to exhaustively assess the performances of each estimator with regard to several biological parameters of the sites under selection (e.g. selection coefficient, dominance model, age of selection). Our analysis highlighted a complementary behavior of these estimators suggesting a possible strategy of merging them in an unified and versatile measure of selective pressure.



A new targeted mutation, Caspa, in the Gnas complex shows hyperactivity and ataxia

Simon Ball1, Christine Williamson1, Charlotte Tibbit1,2, and Jo Peters1

1MRC Harwell, Mammalian Genetics Unit, Oxon, UK

2Current address: MRC Functional Genomics Unit, Oxford, UK


Imprinted genes, by definition, are preferentially expressed from either the maternally or paternally derived allele. They are known to affect growth, survival and behaviour. We report here a new targeted mutation, Caspa, within the Gnas imprinted gene complex. On maternal inheritance, Caspa/+ mice appear chunky have a tail kink or bend and most die before weaning. Notably they are hyperactive, crawling by the day after birth and running by 7-8 days, several days before their wild-type sibs. Ataxia has been seen in some several days after birth, but this is not associated with cerebellar defects. The hyperactivity is reminiscent of that seen in mice with paternal uniparental partial disomy for distal chromosome 2 (PatDp.dist2). In both Caspa/+ and PatDp.dist2, Gnasxl, a paternally expressed transcript in the Gnas complex, is expressed at twice the normal level. We suggest that over expression of Gnasxl, that encodes a variant alpha subunit XLαs of the signalling Gs protein, is the cause of the hyperactivity.




Rat resource and research center

Beth A. Bauer, EC Bryda, CL Franklin, LK Riley, and JK Critser

Department of Veterinary Pathobiology, Research Animal Diagnostic Laboratory, University of Missouri, MO, USA


The Rat Resource and Research Center (RRRC) was established in 2001 with funding from the National Institutes of Health (NIH) National Center for Research Resources (NCRR).  The goals of the RRRC are to (1) provide the biomedical community with a repository and distribution center for valuable rat strains, and (2) to shift the burden for maintaining and distributing unique rat models from investigators to a National Resource Center.  Currently, the RRRC has more than 160 rat lines received through active recruitment of valuable rat models and donations from investigators who have created models. Upon importation into the RRRC, gametes and embryos are cryopreserved to insure against future loss of the model. Recent additions to the repository include a collection of ENU and transposon-generated mutants from the Medical College of Wisconsin.  Also, newly developed rat embryonic stem cell lines have been imported into the RRRC and are available for distribution to the research community. Models in the RRRC are available for distribution as live animals, tissues, or cryopreserved gametes and embryos.  In addition to repository and distribution functions, the RRRC also conducts research.  To date, research efforts by the RRRC have significantly advanced cryobiology and assisted reproductive technologies (ART) for the rat.  Due to high success rates with intracytoplasmic sperm injection (ICSI), the RRRC can use sperm cryopreservation as a cost effective method for banking large collections of single gene mutations and ensuring reliable recovery when models are requested. The RRRC ( is a valuable resource to the community of rat users as it continues to expand the number of accessible rat models and services available to the biomedical community.



Loci on chromosome 2 are associated with na•ve airway hyperresponsiveness in CD-1 outbred mice

David R Beier1, Andrew Kirby2, Mary Prysak1, Mark Daly2, and Emily Cozzi1

1Department of Medicine, Division of Genetics, Brigham and WomenÕs Hospital, Harvard Medical School, Boston MA, USA

2Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA


Asthma is a complex syndrome characterized by episodic and reversible airway obstruction, airway inflammation, and airway hyperresponsiveness. The genetic mechanisms predisposing individuals to asthma and in particular airway hyperresponsiveness (AHR) are largely unknown. Quantitative trait linkage analyses (QTL) have been performed to identify loci associated with naive AHR in a limited number of inbred murine populations. The goal of this study was to uncover loci associated with AHR in a genetically heterogeneous outbred murine population. Outbred CD1 mice were intercrossed to generate F1 progeny from four male/female breeding pairs (N=88). All mice were phenotyped by determining methacholine-induced airway resistance with the invasive flexiVent system.  CD1 F1 mice were genotyped using an Illumina medium density 1440 SNP panel and linkage analysis was carried out using Merlin and GeneHunter software.  The CD1 F1 progeny had a highly variable AHR phenotype (2.2-21.5 cmH2O.s/mL). Linkage analysis of this population was done by inferring parental haplotypes; this identified loci on chromosome 2 as being significantly associated with the AHR phenotype in 2 of the 4 families (additive LOD score = 3). Remarkably, this data is consistent with what we have previously discovered for the A/J and AKR/J inbred hyperresponsive strains. In order to obtain high resolution of the loci identified on chromosome 2, CD1 F1 mice were genotyped on the JAX 600K SNP Diversity Array panel. Analysis of this data may facilitate the identification of specific haplotypes associated with the AHR trait. In summary, we have found a robust distribution and segregation of AHR as a trait in outbred CD1 F1 mice, and identified loci on chromosome 2 that are significantly associated with AHR. Overall, these data highlight that common AHR loci may exist between hyperresponsive murine strains.



High-resolution map and candidate gene analysis for ŽbouriffŽ (ebo), a hair mutation on mouse Chromosome 2 associated with infertility

Fernando Benavides1, J Jaubert2, CJ Perez1, J-L GuŽnet2, J Barrera1, B Cole1, and CJ Conti1

1Department of Carcinogenesis, M.D. Anderson Cancer Center, Science Park-Research Division, Smithville, TX

2UnitŽ de GŽnŽtique Fonctionnelle de la Souris, Institut Pasteur, Paris, France


Forward genetics is a powerful approach to discover genes with pleiotropic effects in the mouse. We previously described ŽbouriffŽ (ebo), a mouse mutation with a hair phenotype and infertility (male and female). This autosomal recessive mutation arose spontaneously in a substrain of BALB/c and was localized on proximal chromosome 2 using an interspecific backcross. Affected ebo/ebo mice show curly vibrissae and wavy hair from day P7, and males have defective spermatogenesis with structural abnormalities of the acrosome that lead to abnormal head shapes and isolated flagellum (Biology of Reproduction 55: 355-363, 1996). New mapping crosses allowed us to narrow down the segment of Chromosome 2 containing ebo to a ~1 Mb interval, between markers D2Mit153 and D2Mit64 (29.27 - 31.20 Mb), a region of homology with human chromosome 9q34. Here, we will summarize the status of our ongoing candidate gene analysis for ebo. Positional candidate genes include Rapgef1, Slc27a4, Coq4, Gle1, Spna2, Zdhhc12, Zer1, Tbc1d13, Ccbl1, and Nup188.





Catrina A Spruce1, John P Flaherty2, Heather E Fairfield1, and David E Bergstrom1

1The Jackson Laboratory, Bar Harbor, ME USA

2New York College of Osteopathic Medicine of New York Institute of Technology, Old Westbury, NY USA


Age-related vestibular decline is a prevalent and increasingly common ailment within the American population. Much of the clinical spectrum is due to the loss and/or displacement of otoconia within the inner ear. Otoconia are proteinaceous particles containing crystalline inorganic calcite. Affixed to underlying stereocilia within the vestibular portion of the inner ear, otoconia act as inertial masses that shift in response to an organismÕs linear movements and to the force of gravity. To better understand the molecular genetics of otoconial deposition and maintenance, we have previously explored the molecular genetic basis for three mouse vestibular mutants named Ņneuroscience mutagenesis facility 333Ó (nmf333), Ņhead tiltÓ (het), and Ņhead slantÓ (hslt), each of which displays the classic phenotype of congenital otoconial agenesis. Together these studies identified Cyba, Nox3, and Noxo1, respectively, as the dysfunction-causative genes. Cyba, in addition to its role in the inner ear, is also a known component of the well-characterized and canonical NADPH oxidase of phagocytic cells. Moreover, Nox3 and Noxo1 are paralogs of the canonical NADPH oxidase components, Cybb and Ncf1. Thus, the emerging picture is that of a Cyba, Nox3 and Noxo1-containing inner ear NADPH oxidase with structural parallels to the well-known phagocytic complex. The critical roll of the inner ear oxidase in otoconiogenesis and proper vestibular function suggests Cyba, Nox3, and Noxo1 as candidate genes potentially involved in human disease- and age-related otoconial loss. Moreover, the Nox3-based NADPH oxidase, as a source of potentially harmful reactive oxygen species, is now under investigation as a possible factor in drug-, age- and noise-related hearing loss. Our presentation will describe recent gene trap and gene targeting studies aimed at elucidating the roles of other NADPH oxidase components in the inner ear.




Heather E Fairfield1, John P Flaherty2, Catrina A Spruce1, Leah R Donahue1, and David E Bergstrom1

1The Jackson Laboratory, Bar Harbor, ME USA

2New York College of Osteopathic Medicine of New York Institute of Technology, Old Westbury, NY USA


Costello Syndrome (CS) is a rare, complex, developmental disorder characterized by a number of features including— failure to thrive, characteristic facies, delay in intellectual development, hypertrophic cardiomyopathy, arrhythmia, and predisposition to both benign and malignant tumors. Past studies identifying gain-of-function mutations in the HRAS gene as the basis of human CS, and strong conservation with the mouse ortholog Hras1, have led to the development of a Gly12Val (G12V) mouse model of CS. To extend animal modeling studies to include various allelic forms of the disease, we are developing five additional mouse models of CS. Using a recombineering-based Ņknock-inÓ approach, we have completed construction of five vectors, each containing a loxP-flanked, neomycin resistance (Neor) selection cassette in intron 1 of Hras1 and site-directed mutations encoding each of the following five pathogenic alleles — G12A encoded by GCA, G12A encoded by GCC, G12S encoded by AGC, G12V encoded by GTA and G12V encoded by GTT. Four of the five strains have been completed creating animal models for the G12AGCA, G12AGCC, G12VGTA and G12VGTT forms of the disease. Suppression of each CS-causative allele by the oppositely transcribed Neor transcript can be relieved by breeding affected mice to inner cell mass- or germline-specific ŅdeleterÓ Cre lines such as Meox2-Cre or Zp3-Cre, respectively. Craniofacial abnormalities like those seen in human patients are clearly evident. Preliminary phenotypic assessment also suggests possible neoplasia as well as cardiac and skin abnormalities. Each strain will be deposited into The Jackson Laboratory Repository. Together, these five models hold the potential for uncovering allelic and codon preference influences on development and neoplasia in CS, identifying CS modifying genes, and dissecting tissue-specific facets of CS using spatially-controlled Cre driver lines.




Laura G Reinholdt, David E Bergstrom, Muriel T Davisson-Fahey, Cathleen M Lutz, Michael Sasner, Steven A Murray, Jeff Lake, Steven Rockwood, Leah Rae Donahue and the MMR team

The Jackson Laboratory, Bar Harbor, ME USA


The application of high-throughput sequencing technologies is revolutionizing the process of mutation detection. By incorporating these technologies, we in The Jackson LaboratoryÕs Mouse Mutant Resource (JAX MMR) are extending our characterization of new spontaneous mutant strains to the point of causative gene identification. For over fifty years, the mission of the MMR (and its predecessors) has been to provide mouse models of human genetic illness to the scientific community. Through our Mouse Phenodeviant Search process, atypical mice are identified from among the millions of inbred mice produced yearly at JAX and isolated. This not only ensures genetic stability within the originating strain; but also, provides a pool of potential spontaneous mutant mice for further characterization. After colonies are established, heritability is proven, and the modes of inheritance are determined; subchromosomal locations for each mutant locus are established by backcross or intercross in conjunction with SSLP or SNP genotyping. In place of Sanger-based sequencing of promising candidate genes from select mutant strains, interval-specific array capture and Illumina GAIIx-based high-throughput sequencing are now being incorporated to broaden the scope of mutation detection and dramatically shorten the time to causative gene identification. A general phenotypic assessment, cryopreservation of embryos or gametes, and archival of mutant DNA completes the characterization. Findings for each mutant strain are disseminated through the scientific literature or on the MMR website at Additional projects are underway to incorporate whole-exome approaches to spontaneous/induced mutation discovery, reducing the need for genetic mapping. The presentation will enumerate the many advantages of studying spontaneous mutant mice and summarize current resource offerings.




Elizabeth C Bryda1, Nathan T Johnson1, Cynthia L Besch-Williford1, Kevin K Ohlemiller2, and Richard J Bram3

1Department of Veterinary Pathobiology, Research Animal Diagnostic Laboratory, College of Veterinary Medicine, University of Missouri, Columbia, MO, USA

2Central Institute for the Deaf at Washington University, Department of Otolaryngology, Washington University School of Medicine, St. Louis, MO, USA

3Department of Immunology, Mayo Clinic College of Medicine, Rochester, MN, USA


Calcium modulating cyclophilin ligand (CAML) is a ubiquitously expressed cytoplasmic protein which appears to play a role in embryonic and thymocyte development, EGFR and LCK signaling, and chromosomal stability.  In previous studies, we demonstrated that there is a protein –protein interaction between CAML and the cytoplasmic region of Cadherin23.  This led us to speculate that CAML might be important in the inner ear and  play a role in the development and/or function of hair cells. To enable spatially and temporally controlled Caml gene expression, a mouse line was generated that allows knockout of Caml expression specifically in the inner ear upon administration of tamoxifen. Using this mouse model, tamoxifen was administered immediately after birth to neonates or daily to pregnant females from day E10.5 to E15 to assess the effect of loss of CAML during postnatal or embryonic development respectively.  Hearing in treated animals was tested by auditory brainstem response (ABR) analysis. The presence of hair cells was evaluated via histological analysis of the inner ears of treated mice.  To date, there is evidence that lack of Caml expression in the inner ear leads to deafness.  Elucidating a role for CAML in the inner ear will aid our understanding of the molecular pathways important for auditory development and function.



Interesting phenotypes found as part of the infection challenge in the Wellcome Trust Sanger InstituteÕs Mouse Genetics Programme

Simon Clare, Leanne Kane, Lynda Mottram, Jacqui White, Ramiro Ramirez – Solis, and Gordon Dougan

All the Mouse genome project team at the Sanger Institute

Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Cambridgeshire, UK


As part of the Wellcome Trust Sanger InstituteÕs Mouse Genetics Programme all mutant mouse lines generated in this high throughput programme are challenged with Salmonella Typhimurium an intracellular pathogen which induces a systemic disease and Citrobacter rodentium a natural mouse pathogen which forms attaching and effacing lesions on the surface of the gastrointestinal lumen (details of the challenges can be found on the poster  ŅPhenotyping of knockout mice using bacterial pathogens as part of the Wellcome Trust Sanger InstituteÕs Mouse Genetics ProgrammeÓ). To date we have identified 18 genes which contribute to controlling the susceptibility to bacterial infection out over 250 knockout mice lines phenotyped so far. These phenotypes include hits in novel genes as well as gene of known function and range from severe phenotypes, were the mice succumb to infection rapidly, too more subtle phenotypes. Here we would like to show a selection of the data generated as part of this programme and details of how to freely access all the data and resources from the programme as a whole.




The expression of septin 9 in human breast cancer

Diana Connolly1, Zhixia Yang1, Elias T. Spiliotis2, Maria Castaldi3, Nichelle Simmons3, Pascal Verdier-Pinard4, and Cristina Montagna

1Department of Genetics, Albert Einstein College of Medicine of Yeshiva University, Bronx, NY, USA

2Department of Biology, Drexel University, Philadelphia, PA, USA

3Department of Surgery, Jacobi Medical Center, Bronx, NY, USA

4INSERM UMR 911 CR02, Aix-Marseille UniversitŽ, Marseille, France


Septins are a family of GTPases known for their involvement in various cellular processes and interactions with the cytoskeleton.  Septin 9 (SEPT9) in particular, is one of the fourteen members of this family that is of particular interest because of its contribution to breast tumorigenesis. Our group identified SEPT9 as a novel oncogene that is amplified and over-expressed in breast cancer. Alteration of SEPT9 expression has also been implicated in ovarian carcinoma and results in cell cycle arrest when down-regulated. The study of SEPT9 is complex due to the potential ability of this gene to encode 18 different isoform transcripts and at least 5 different polypeptides. Studies show that SEPT9 isoforms are differentially expressed between normal and tumor breast cells, but their relation to stages of breast cancer development and breast cancer subtypes is still unclear.  Our group has approached this question by analyzing isoform expression at the DNA, RNA and protein levels.  Using mouse models for breast cancer progression, and human tumor and matching adjacent normal breast tissue, we have detected gene amplification and expression changes of the SEPT9 isoforms among the tissue types. For the first time we could show that pre-malignant cells have a SEPT9 expression profile that distinguishes them from both normal breast and advanced adenocarcinomas. Moreover, we found that the differential expression of at least one isoform (SEPT9_v3) was due to epigenetic regulation via the methylation status of an alternative promoter region.  Our data suggest that other alternative promoters within this gene could also be regulated by DNA methylation and thus dramatically affect the expression of SEPT9 isoforms in tumorigenesis. These findings are supported by the establishment of a diagnostic SEPT9 DNA methylation assay for detection of colorectal cancer in peripheral blood, and furthermore indicate that the amplification and differential expression of SEPT9 isoforms may represent a potential molecular signature with functional and diagnostic significance for breast tumor progression and early detection.




David DeSantis1, Michelle Pritchard3, Laura Nagy3, Joseph Nadeau2, and Colleen M Croniger1

1Departments of Nutrition, 2Department of Genetics, School of Medicine, Case Western Reserve University, Cleveland, Ohio, USA

3Department of Gastroenterology, Department of Pathobiology, Cleveland Clinic, Cleveland, Ohio, USA


Liver disease is one of the serious complications associated with either chronic excessive alcohol consumption or calorie consumption. Complications can range from steatosis to hepatitis and eventually fibrosis and cirrhosis.  Pathological evolution of alcoholic steatohepatitis (ASH) and non-alcoholic steatohepatitis (NASH) are very similar yet only 20-50% of  these patients develop cirrhosis. The mechanism for progression to fibrosis in these patients is presently unknown. Current nutritional and transgenic models of hepatic steatosis replicate preconditions for ASH and NASH, but fail to represent the multifactorial pathogenesis of these diseases. To identify susceptibility genes to fibrosis, we have analyzed the response to high fat high sucrose diet (HFHS) in chromosome substitution strains (CSS) developed by Dr. Joseph Nadeau (Case Western Reserve University). CSS strains have one chromosome