Functional characterization of the trout insulin promoter: implications for fish as a favorable model of pancreas development.
The complex anatomy of the mammalian pancreas, in which the endocrine cells are grouped in islets dispersed among the predominant exocrine component, has hampered study of the molecular events governing the development of pancreatic cell lineages. To investigate whether fish may provide relevant, complementary models of pancreas development, we characterized the trout insulin (tINS) promoter and its molecular interactions with PDX1, a key transcriptional and developmental factor of the mammalian pancreas. Transfection of a luciferase reporter plasmid containing the 280 bp 5'-flanking region of the tINS gene resulted in strong activity in mammalian pancreatic beta cells but not in CHO or pituitary cells. Footprinting assays and cotransfection experiments indicated that mammalian PDX1 binds to and activates the tINS promoter. By microinjecting plasmids to fertilized zebrafish eggs, we showed that the expression of mouse PDX1 is capable of activating the co-injected tINS promoter plasmid in most cell types of the 24-h zebrafish embryo. The conserved role of PDX1 in vertebrate insulin gene regulation opens the possibility to exploit fish models in the study of pancreas development.
Argenton
F
F
Dipartimento di Biologia, Università di Padova, Padua, Italy.
Walker
M D
MD
Colombo
L
L
Bortolussi
M
M
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006224:Cricetinae; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins; D007328:Insulin / Q000235:genetics*; D007515:Islets of Langerhans / Q000166:cytology / Q000254:growth & development*; D051379:Mice; D008969:Molecular Sequence Data; D010902:Pituitary Gland / Q000166:cytology; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051381:Rats; D015534:Trans-Activators / Q000378:metabolism*; D014162:Transfection; D014337:Trout / Q000235:genetics*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics
D018398:Homeodomain Proteins; D007328:Insulin; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein