Translocation of protein kinase C by halothane in cholinergic cells.
Protein kinase C (PKC) is a signal transducing enzyme that is an important regulator of multiple physiologic processes and a potential molecular target for volatile anaesthetic actions. However, the effects of these agents on PKC activity are not yet fully understood. Volatile anaesthetics increase intracellular calcium concentration ([Ca(2+)](i)) in a variety of cells, thus their effects on PKC activity may be indirect due to [Ca(2+)](i) increase. Alternatively, the anaesthetics could directly stimulate PKC activity. In order to distinguish these two possibilities in intact cells, we used a fully functional green fluorescent protein conjugated PKCbetaII (GFP-PKCbetaII) and confocal microscopy to evaluate the dynamic redistribution of PKC in living SN56 cells, a cholinergic cell line, in response to halothane. Halothane induced PKC translocation in SN56 cells transfected with GFP-PKCbetaII. This effect was not suppressed by dantrolene, a drug that blocks halothane-induced Ca(2+) release from intracellular stores in these cells. These findings indicate that halothane induces PKC translocation in SN56 cells independently of its ability to release calcium from internal stores.
Departamento de Cirurgia, Faculdade de Medicina da UFMG, Belo Horizonte-Minas Gerais, Brazil. renatogomez@ig.com.br
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RS
Gomez
J
J
Barbosa
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C
Guatimosim
A R
AR
Massensini
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MV
Gomez
M A M
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Prado
D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002799:Cholinergic Fibers / Q000187:drug effects / Q000201:enzymology*; D003620:Dantrolene / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D006221:Halothane / Q000494:pharmacology*; D007202:Indicators and Reagents / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal; D009125:Muscle Relaxants, Central / Q000494:pharmacology; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D064546:Protein Kinase C beta
D018685:Anesthetics, Inhalation; D007202:Indicators and Reagents; D007527:Isoenzymes; D008164:Luminescent Proteins; D009125:Muscle Relaxants, Central; D049452:Green Fluorescent Proteins; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D003620:Dantrolene; D002118:Calcium; D006221:Halothane