The role of protein kinase C in the regulation of serotonin-2A receptor expression.
We have investigated in C6 glioma cells the involvement of protein kinase C (PKC) in the regulation of serotonin-(2A) receptor (5-HT(2A) receptor) expression by agonist treatment. Comparison of the time-courses of agonist-induced downregulation of receptor number and mRNA indicate that a decrease in the number of 5-HT(2A) receptor binding sites in response to serotonin (5-HT) treatment is preceded by a decrease in 5-HT(2A) receptor mRNA. This decrease in 5-HT(2A) receptor mRNA as a result of agonist exposure was not due to a change in the stability or half-life of the transcript. Pretreatment of cells with the PKC inhibitor bisindolylmaleimide blocked the decrease in 5-HT(2A) receptor mRNA levels, and attenuated the down-regulation of 5-HT(2A) receptor binding sites induced by treatment with 5-HT. Experiments performed with the PKC inhibitors calphostin C and Gö 6976 confirmed that PKC was involved in the regulation of 5-HT(2A) receptor mRNA by agonist and implicate the conventional subgroup of PKC isoforms. Western blot analysis, using isoform-specific anti-PKC antibodies showed that under our culture conditions C6 glioma cells express the conventional isoforms PKC alpha, PKC gamma, as well as the novel isoforms PKC delta, PKC epsilon, and the atypical isoforms PKC lambda and PKC iota. Upon treatment with 5-HT for 10 min levels of the conventional isoforms PKC alpha and PKC gamma increased in the nuclear fraction. Taken together, our results implicate PKC alpha and/or PKC gamma in the regulation of 5-HT(2A) mRNA receptor and binding sites in response to agonist treatment.
Anji
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A
Department of Pharmacology, University of Texas Health Science Center, San Antonio, Texas, USA.
Sullivan Hanley
N R
NR
Kumari
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Hensler
J G
JG
D000818:Animals; D001665:Binding Sites; D002227:Carbazoles / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects*; D004790:Enzyme Induction / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005910:Glioma / Q000473:pathology; D007211:Indoles / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D051379:Mice; D009281:Naphthalenes / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis; D012334:RNA, Neoplasm / Q000096:biosynthesis; D044402:Receptor, Serotonin, 5-HT2A; D011985:Receptors, Serotonin / Q000096:biosynthesis* / Q000235:genetics; D012701:Serotonin / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured / Q000201:enzymology
D002227:Carbazoles; D007211:Indoles; D008301:Maleimides; D009281:Naphthalenes; D009363:Neoplasm Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D044402:Receptor, Serotonin, 5-HT2A; D011985:Receptors, Serotonin; C081021:Go 6976; D012701:Serotonin; D011493:Protein Kinase C; C058819:calphostin C; C088060:bisindolylmaleimide