Expression of calcium channel mRNAs in rat pancreatic islets and downregulation after glucose infusion.
Recent studies have shown that two different voltage-dependent Ca2+ channels are expressed in pancreatic islets, the beta-cell/neuroendocrine-brain and the cardiac subtypes. The effects of chronic hyperglycemia on the levels in pancreatic islets of the mRNAs encoding the alpha 1-subunits of the beta-cell and cardiac subtype Ca2+ channels were studied in rats made hyperglycemic by infusion of glucose for 48 h. A competitive reverse transcriptase-polymerase chain reaction procedure was used to obtain quantitative data on the levels of these two transcripts in islets obtained from individual rats. The quantitative polymerase chain reaction data indicate that the levels of mRNA encoding the alpha 1-subunit of the beta-cell Ca2+ channel are 2.5-fold greater than those for the cardiac subtype. The levels of beta-cell Ca2+ channel mRNA were 72.9% lower in the glucose-infused animals when compared with the saline-infused animals (P < 0.005) and those of the cardiac channel were 72.1% lower in the animals infused with glucose (P < 0.02). In contrast, glucose infusion resulted in a twofold increase in insulin mRNA levels and did not significantly alter levels of beta-actin mRNA. In situ hybridization studies revealed that the mRNAs for these two Ca2+ channels are expressed at higher levels in normal rat islets than in the surrounding acinar tissue, which suggests that the observed changes in mRNA levels occur within cells of the pancreatic islet. To assess the possible functional consequences of this reduction in expression of mRNA for the Ca2+ channels, the insulin secretory responses of perfused pancreases to the Ca2+ channel agonist Bay K8644 were studied.(ABSTRACT TRUNCATED AT 250 WORDS)
Iwashima
Y
Y
Department of Medicine, University of Chicago, IL 60637.
Pugh
W
W
Depaoli
A M
AM
Takeda
J
J
Seino
S
S
Bell
G I
GI
Polonsky
K S
KS
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000199:Actins / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D016375:Antisense Elements (Genetics); D001483:Base Sequence; D015220:Calcium Channels / Q000187:drug effects / Q000235:genetics*; D004247:DNA; D015870:Gene Expression / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology*; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D009838:Oligodeoxyribonucleotides; D010477:Perfusion; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection
D000199:Actins; D016375:Antisense Elements (Genetics); D015220:Calcium Channels; D007328:Insulin; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D004247:DNA; D005947:Glucose