Cyclic AMP stimulates the gene expression of a non-selective cation channel, mNSC1, in pancreatic beta-cell line, MIN6

Mol Cell Endocrinol. 2000 Feb 25;160(1-2):165-71. doi: 10.1016/s0303-7207(99)00214-2.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cyclic AMP / pharmacology*
  • DNA Primers / genetics
  • Gene Expression / drug effects
  • Half-Life
  • Histone-Lysine N-Methyltransferase
  • Ion Channels / genetics*
  • Islets of Langerhans / drug effects*
  • Islets of Langerhans / metabolism*
  • Mice
  • Promoter Regions, Genetic
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism

Substances

  • DNA Primers
  • Ion Channels
  • RNA, Messenger
  • Cyclic AMP
  • Histone-Lysine N-Methyltransferase
  • Nsccn1 protein, mouse