To investigate the role of the Sry/hydroxymethylglutaryl box (Sox) transcription factors in the development of the pancreas, we determined the expression pattern of Sox factors in the developing mouse pancreas. By RT-PCR, we detected the presence of multiple Sox family members in both the developing pancreas and mature islets and then focused on two factors, Sox2 and Sox4. The expression field of Sox2, which plays a role in the maintenance of some stem cell populations, included the developing duodenum, but Sox2 was specifically excluded from the pancreatic buds. In contrast, Sox4 was detected broadly in the early pancreatic buds and eventually became restricted to the nuclei of all islet cells in the adult mouse. Mice homozygous for a null mutation of the sox4 gene showed normal pancreatic bud formation and endocrine cell differentiation up to embryonic day 12.5. Beyond that date, cultured pancreatic explants lacking sox4 failed to form normal islets. Instead, a markedly reduced number of endocrine cells were found scattered through the explant. We show here that several Sox transcription factors are expressed in the developing pancreas and in the islet, and that one of these factors, Sox4, is required for the normal development of pancreatic islets. Diabetes 54:3402-3409, 2005 S cattered through the exocrine pancreas, the islets of Langerhans are highly organized clusters of endocrine cells comprised of four distinct cell types: the glucagon-secreting ␣-cells, the insulinsecreting ␤-cells, the somatostatin-secreting ␦-cells, and the pancreatic polypeptide-secreting cells. Because of the essential role of the islet hormones in energy metabolism, defects in the development, maintenance, or function of the endocrine pancreas have serious consequences, including diabetes.The pancreas forms from the endoderm in the region of the foregut/midgut junction and is first visible in mice at embryonic day 9.5 (e9.5) (1). The first endocrine cells of the pancreas appear at around e9.5 in the dorsal pancreatic bud. These cells express glucagon; a few insulinexpressing cells appear ϳ1 day later. At ϳe13-14, the pancreas undergoes a distinct change termed the secondary transition, characterized by the appearance of ductal cells, exocrine cells, and delta cells and a rapid expansion of the insulin-expressing cells. The endocrine cells found in the pancreas before the secondary transition have clear differences from mature islet cells and may result from developmental pathways that are distinct from those that produce the more mature endocrine cells that arise following the secondary transition (2-6). By e18, the first pancreatic polypeptide-expressing cells appear, and the endocrine cells organize into distinct islets with the ␤-cells forming the central core. Around the same time, ␤-cell neogenesis declines, but the simultaneous onset of ␤-cell replication continues to expand the ␤-cell population and enlarge the forming islets (7).This process of endocrine cell determination and differentiation depends on the proper sequentia...

Recent genetic analysis of the Drosophila dachshund (dac) gene has established that dac encodes a novel nuclear protein that is involved in both eye and leg development. In the Drosophila eye, dac expression appears to be controlled by the product of the eyeless/Pax6 gene. In order to analyze the Pax6 pathway in vertebrates we have isolated and characterized the cDNA and genomic clones corresponding to the human and mouse homologues of Drosophila dac. A full-length human cDNA encoding dachshund (DACH) encodes the 706 amino acids protein with predicted molecular weight of 73 kDa. A 109 amino acid domain located at the N-terminus of the DACH showed significant sequence and secondary structure homologies to the ski/sno oncogene products. Northern blot analysis found human DACH predominantly in adult kidney, heart, and placenta, with less expression detected in the brain, lung, skeletal muscle and pancreas. A panel of human cell lines was studied and most notably a large proportion of neuroblastomas expressed DACH mRNA. Mouse Dach encodes a protein of 751 amino acids with predicted molecular weight of 78 kDa that is 95% identical to the human DACH. RNase protection analysis showed the highest Dach mRNA expression in the adult mouse kidney and lung, whereas lower expression was detected in the brain and testis. RT/PCR analysis readily detected Dach mRNA in the adult mouse cornea and retina. Dach mRNA expression in the mouse E11.5 embryo was observed primarily in the fore and hind limbs, as well as in the somites.

All metazoans use insulin to control energy metabolism, but they secrete it from different cells: neurons in the central nervous system in invertebrates and endocrine cells in the gut or pancreas in vertebrates. Despite their origins in different germ layers, all of these insulin-producing cells share common functional features and gene expression patterns. In this study, we tested the role in insulin-producing cells of the vertebrate homologues of Dachshund, a transcriptional regulator that marks the earliest committed progenitors of the neural insulin-producing cells in Drosophila. Both zebrafish and mice expressed a single dominant Dachshund homologue in the pancreatic endocrine lineage, and in both species loss of this homologue reduced the numbers of all islet cell types including the insulin-producing β-cells. In mice, Dach1 gene deletion left pancreatic progenitor cells unaltered, but blocked the perinatal burst of proliferation of differentiated β-cells that normally generates most of the β-cell mass. In β-cells, Dach1 bound to the promoter of the cell cycle inhibitor p27Kip1, which constrains β-cell proliferation. Taken together, these data demonstrate a conserved role for Dachshund homologues in the production of insulin-producing cells.

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