Glucose-stimulated expressed sequence tags from rat pancreatic islets.
Available evidence suggests that glucose, the most potent physiologic insulin secretagogue, capacitates glucose-induced insulin secretion by stimulating synthesis of various proteins in the beta cell. To obtain more clues about proteins that might be involved in insulin secretion, rat pancreatic cDNA libraries were screened by differential hybridization for non-preproinsulin transcripts that were increased when pancreatic islets were cultured for 1 day at a high (20 mM) versus a low concentration (1 mM) of glucose. More than 100,000 pfu were initially screened. After repeated rescreening, 33 transcripts were 1-3-fold higher in the presence of the high glucose. For comparison, preproinsulin transcripts were 4-8-fold higher at the high concentration of glucose. The sequences of 12 clones were > or = 85% similar to published sequences. These included annexin, calbindin, protein kinase C receptor, the G protein beta subunit, the guanyl cyclase A/atrial natriuretic peptide receptor and the serotonin 5HT-2 receptor. As previously reported, ferritin H chain transcripts were discovered to be 3-6-fold higher in the presence of the high glucose (MacDonald et al., FASEB J. 8, 777-781, 1994). Unidentified glucose responsive clones have been assigned GenBank accession numbers N55606-N55636, N65938 and N65939. The results implicate the proteins encoded by these mRNAs in insulin secretion.
MacDonald
M J
MJ
University of Wisconsin Children's Diabetes Center, Madison 53706, USA. mjmacdon@facstaff.wisc.edu
D000818:Animals; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000494:pharmacology*; D007515:Islets of Langerhans; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic / Q000187:drug effects*
D018076:DNA, Complementary; D005947:Glucose