International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


POSTER 56 - A TRANSCRIPTOMIC APPROACH TO STUDY THE REGULATORY FUNCTION OF HFE, THE HEMOCHROMATOSIS GENE PRODUCT, IN MOUSE MODELS

E Abgueguen
UMR 6061 CNRS

Fergelot P , Ferran H, Sachot S, Orhant M, Gicquel I, Mottier S, Soriano N, Mosser J
UMR 6061 CNRS, Faculté de Médecine

We aimed to precise the regulatory mechanisms involved in intestinal iron absorption, which dysfunction leads to anemia or iron overload. For this purpose we used transgenic models of targeted expression of HFE in gut epithelium, HFE -/- (Bahram et al., 1999) and HFE knock-out bred with HFE transgenic mice (Fergelot et al., 2002). These mice provide integrative models to study the role of HFE1, the gene mutated in hereditary hemochromatosis, on iron transfer during enterocyte differentiation. We are now developing a transcriptomic approach to correlate the pattern of expression of cryptic and mature enterocytes with their HFE status. Transcriptomic analysis was initiated on intestinal cell lines to select a subset of genes that could be differentially expressed in varied biological conditions. cDNAs clones obtained after subtractive suppressive hybridization (SSH) between undifferentiated and differentiated Caco2 intestinal cell line were spotted and hybridized with mouse mRNA from cryptic cells or differentiated enterocytes. Due to a considerable variation of differentially expressed clones with strigency conditions, we built cDNA microarrays dedicated to murine tissues. SSH between the whole duodenal cells of HFE-/- mice and transgenic mice generated 100 up-regulated cDNA clones in HFE transgenic mice and 180 up-regulated cDNA clones in HFE-/- mice. Expression profiles of cryptic or mature enterocytes from HFE-/-, normal and the different HFE transgenic lines are analysed with the Genepix Pro 4.0 sofware (Axon). Statistical analysis is performed according to Dudoit and colleagues (Dudoit et al., 2000). We are also currently implementing an ontology database to improve the clustering of differentially expressed genes.


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