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Journal of Molecular Endocrinology (2006) 37, 301-316    DOI: 10.1677/jme.1.02096
© 2006 Society for Endocrinology
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The effect of neurogenin3 deficiency on pancreatic gene expression in embryonic mice

Andreas Petri1,2,*, Jonas Ahnfelt-Rønne2,*, Klaus Stensgaard Frederiksen1, David George Edwards3, Dennis Madsen4, Palle Serup2, Jan Fleckner1 and R Scott Heller2

1 Department of Molecular Genetics, Novo Nordisk A/S, DK-2880 Bagsværd, Denmark
2 Department of Developmental Biology, Hagedorn Research Institute, DK-2820 Gentofte, Denmark
3 Department of Biostatistics, Novo Nordisk A/S, DK-2880 Bagsværd, Denmark
4 Department of Scientific Computing, Novo Nordisk A/S, DK-2760 Måløv, Denmark

(Requests for offprints should be addressed to R S Heller; Email: shll{at}hagedorn.dk)

* (A Petri and J Ahnfelt-Rønne contributed equally to this work)

To understand the molecular mechanisms regulating pancreatic endocrine development and function, pancreatic gene expression was compared between Ngn3-deficient mice and littermate controls on embryonic days 13 and 15. Microarray analysis identified 504 genes with significant differences in expression. Fifty-two of these showed at least twofold reduction in Ngn3 knockouts compared to controls. Many of them were previously described to be involved in endocrine development and function. Among the genes not previously characterized were Rhomboid veinlet-like 4, genes involved in tetrahydrobiopterin biosynthesis and the Iroquois-type homeobox gene Irx1, the latter was selected for further investigation. In situ hybridisation demonstrated that two Iroquois genes, Irx1 and Irx2, were expressed in pancreatic endoderm of wild-type, but not Ngn3 mutant embryos. Furthermore, ectopic Ngn3 induced prominent Irx2 expression in chicken endoderm. Co-labelling established that Irx1 and Irx2 mRNA is located to glucagon-, but not insulin- or somatostatin-producing cells in mice and chicken. These data suggest that Irx1 and Irx2 serve an evolutionary conserved role in the regulation of {alpha}-cell-specific gene expression.




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