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Cyclic AMP stimulates the gene expression of a non-selective cation channel, mNSC1, in pancreatic beta-cell line,
1Department of Pharmacology, Jichi Medical School, 3311-1 Yakushiji, Minamikawachi, Kawachi, Tochigi, Japan.
Abstract:
Mouse non-selective cation channel 1 (mNSC 1) cDNA from mouse pancreatic beta-cell line, MIN6, have recently been cloned. Since the number of non-selective cation channel in pancreatic duct cells has been reported to increase 9-fold in 5 h incubation with cAMP, the effect of cAMP on the gene expression of mNSC1 in MIN6 cells was examined. Dibutyryl cAMP (db-cAMP) was shown to increase the level of the mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The copy number of the mRNA was increased 4-fold in 6 h incubation with db-cAMP by competitive PCR. Western blot analysis also indicated a 4-fold increase in the quantity of the newly synthesized protein in 9 h incubation with db-cAMP. Experiments with 5'-flanking region and with a transcriptional inhibitor suggested that db-cAMP affected transcription, and protected the mRNA from its degradation as well. It is concluded that the expression of mNSC1 is indeed increased by cAMP in the pancreatic beta-cells.
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