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Submitted on April 19, 2002
Accepted on July 17, 2002
1 The Department of Medicine, Banting and Best Diabetes Centre, Toronto General Hospital, University of Toronto, Toronto, Ontario, M5G 2C4
* To whom correspondence should be addressed. E-mail: d.drucker{at}utoronto.ca.
Tissue-specific proglucagon gene transcription is achieved through combinations of transcription factors expressed in pancreatic A cells and enteroendocrine L cells of the small and large intestine. Cell transfection and electrophoretic mobility shift assay (EMSA) experiments previously identified Pax-2 as a regulator of islet proglucagon gene expression. We examined whether Pax-2 regulates gut proglucagon gene expression using enteroendocrine cell lines and Pax21NEU mutant mice. Immunoreactive Pax-2 was detected in STC-1 enteroendocrine cells and Pax-2 activated proglucagon promoter activity in transfected BHK and GLUTag cells. Pax-2 antisera diminished the formation of a Pax-2-G3 complex in EMSA studies using nuclear extracts from islet and enteroendocrine cell lines. Surprisingly, Pax-2 mRNA transcripts were not detected by RT-PCR in RNA isolated from adult rat pancreas, rat islets, E19 or adult murine pancreas and gastrointestinal tract. Furthermore, E19 or neonatal day 1 Pax21NEU mice exhibited normal islet A cells and gut endocrine L cells, and no decrement in pancreatic or intestinal glucagon gene expression. These findings demonstrate that Pax-2 is not essential for the developmental formation of islet A or gut L cells and does not play a role in the physiological control of proglucagon gene expression in vivo.
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