Cyclic AMP stimulates the gene expression of a non-selective cation channel, mNSC1, in pancreatic beta-cell line, MIN6.
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.
Satoh
J
J
Department of Pharmacology, Jichi Medical School, 3311-1 Yakushiji, Minamikawachi, Kawachi, Tochigi, Japan.
Kutsuwada
K
K
Ohki
G
G
Imai
M
M
Kobayashi
M
M
Suzuki
M
M
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D000242:Cyclic AMP / Q000494:pharmacology*; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D006207:Half-Life; D011495:Histone-Lysine N-Methyltransferase; D007473:Ion Channels / Q000235:genetics*; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D017931:DNA Primers; D007473:Ion Channels; D012333:RNA, Messenger; D000242:Cyclic AMP; D011495:Histone-Lysine N-Methyltransferase; C110219:Nsccn1 protein, mouse