International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


POSTER 137 - FRESH AND CRYOPRESERVED SPERM QUALITY, DNA INTEGRITY, AND IN VITRO FERTILITY IN TWO HYBRID STRAINS OF N-ETHYL-N-NITROSOUREA (ENU) MICE

Cengiz Yildiz CY, Craig Fleming FC, Amanda Cao AC, Colin McKerlie CM

1 The Hospital for Sick Children, Toronto, Canada, 2 The Hospital for Sick Children, Toronto, Canada, 3 The Hospital for Sick Children, Toronto, Canada, 4 The Hospital for Sick Children, Toronto, Canada

Background: Efficient collection, freezing, archiving, and re-derivation of sperm are essential components for random mutagenesis in the mouse. The purpose of this study was to determine the effect(s) of a triple dose of ENU (225 or 300 mg/kg total dose) on fresh and frozen-thawed sperm quality, spermatozoa DNA integrity, and unassisted in vitro fertility rates in C3B6G1 and B6129S1G1 mice that are generated, used, and maintained in the mutagenesis program at the Centre for Modeling Human Disease in Toronto.

Methods:  Sperm was collected from C3B6G1 and B6129S1G1 control and ENU mice. Sperm assessment parameters included progressive motility, plasma membrane integrity, concentration, membrane function integrity, acrosome integrity, and DNA integrity. To assess the effect(s) of oocyte donor strain, the in vitro fertilization rate, embryo culture, and live-born offspring rates from fresh and frozen sperm from G1 and age-matched control wild-type hybrid mice were determined when either of the parental or hybrid strain females were used as oocyte donors.

Results: There were no significant differences in fresh or frozen sperm quality parameters between wild-type and C3B6G1 or B6129S1G1 mice (P>0.05) with these doses of ENU. However, there were significant differences in the in vitro fertilization rate, embryo culture rate, and live-born offspring rates when the background strain of the oocyte donor did not match the sperm donor. Representative data will be presented to confirm that background strain of sperm or oocyte donor is an important consideration and can affect the efficiency of derivation of mice from frozen mouse sperm.

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