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Developmental Biology of the Psammomys obesus Pancreas: Cloning and Expression of the Neurogenin-3 Gene

Louise Vedtofte, Thóra B. Bödvarsdóttir, Allan E. Karlsen and R. Scott Heller

Department of Developmental Biology, Hagedorn Research Institute, Gentofte, Denmark (LV,RSH); Department of Diabetes Pharmacology (TBB) and Department of Insulin, Incretin, and Islet Biology (AEK), Novo Nordisk, Måløv, Denmark; and Institute for Clinical Science, University of Lund, Lund, Sweden (AEK)

Correspondence to: R. Scott Heller, PhD, Hagedorn Research Institute, DK 2820 Gentofte, Denmark. E-mail: shll{at}hagedorn.dk

The desert gerbil Psammomys obesus, an established model of type 2 diabetes (T2D), has previously been shown to lack pancreatic and duodenal homeobox gene 1 (Pdx-1) expression. Pdx-1 deficiency leads to pancreas agenesis in both mice and humans. We have therefore further examined the pancreas of P. obesus during embryonic development. Using Pdx-1 antisera raised against evolutionary conserved epitopes, we failed to detect Pdx-1 immunoreactivity at any time points. However, at E14.5, Nkx6.1 immunoreactivity marks the nuclei of all epithelial cells of the ventral and dorsal pancreatic buds and the only endocrine cell types found at this time point are glucagon and PYY. At E18.5 the pancreas is well branched and both glucagon- and ghrelin-positive cells are scattered or found in clusters, whereas insulin-positive cells are not found. At E22.5, the acini of the exocrine pancreas are starting to mature, and amylase and carboxypeptidase A immunoreactivity is found scattered and not in all acini. Ghrelin-, glucagon-, PYY-, gastrin-, somatostatin (SS)-, pancreatic polypeptide (PP)-, and insulin-immunoreactive cells are found scattered or in small groups within or lining the developing ductal epithelium as marked by cytokeratin 19. Using degenerate PCR, the P. obesus Neurogenin-3 (Ngn-3) gene was cloned. Nucleotide and amino acid sequences show high homology with known Ngn-3 sequences. Using specific antiserum, we can observe that Ngn-3-immunoreactive cells are rare at E14.5 but readily detectable at E18.5 and E22.5. In conclusion, despite the lack of detection of Pdx-1, the P. obesus pancreas develops similarly to Muridae species, and the Ngn-3 sequence and expression pattern is highly conserved in P. obesus. (J Histochem Cytochem 55:97–104, 2007)

Key Words: Psammomys obesus • neurogenin-3 • type 2 diabetes • developmental biology • glucagon • insulin • pancreas • sand rat

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