Acetylcholine agonists stimulate mitogen-activated protein kinase in oligodendrocyte progenitors by muscarinic receptors.
Oligodendrocytes, the myelin-producing cells of the central nervous system, express muscarinic acetylcholine receptors (mAChR). Activation of this neurotransmitter receptor by the stable acetylcholine analog carbachol (CCh) triggers transducing events, modulating c-fos expression and cellular proliferation. To elucidate the signal transduction pathways involved in the transmission of these cellular events, we examined the ability of CCh to activate mitogen-activated protein kinase (MAPK) in primary cultures of oligodendrocyte progenitors prepared from newborn rat brain. CCh produced a concentration- and time-dependent increase in MAPK activity (predominantly the p42mapk or ERK2) as determined by in-gel MBP kinase assays. Using the non-selective muscarinic antagonist atropine we determined that MAPK-activation by CCH is mediated by muscarinic receptors. In the presence of PD098059, a specific inhibitor of MAPK kinase (MEK), MAPK activity was blocked. Similarly, the presence of extracellular calcium was required for CCh-mediated MAPK activation. To further elucidate the mechanisms involved in MAPK activation by CCh, the role of PKC was studied. In cells in which protein kinase had been downregulated by chronic treatment with 12-O-tetradecanoylphorbol 13-acetate (TPA), the effect of carbachol on MAPK activation was maintained. In contrast, the response to CCh was blocked by the PKC inhibitors H7 and bisindolylmaleimide GF109203X. Our results suggest that MAPK is implicated in the transmission of the signal for mACh receptors and involves a TPA-insensitive PKC pathway. Further work is required to define the upstream and downstream events which result in CCh-mediated MAPK activation and proliferation of oligodendrocyte progenitors.
Larocca
J N
JN
Department of Neurology, Albert Einstein College of Medicine, Bronx, New York 10461, USA. Larocca@aecom.yu.edu
Almazan
G
G
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D001285:Atropine / Q000494:pharmacology; D001921:Brain / Q000166:cytology / Q000502:physiology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology*; D002478:Cells, Cultured; D018678:Cholinergic Agents / Q000494:pharmacology*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D009836:Oligodendroglia / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D051381:Rats; D017207:Rats, Sprague-Dawley; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects; D013234:Stem Cells / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D018678:Cholinergic Agents; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007211:Indoles; D008301:Maleimides; D011976:Receptors, Muscarinic; D015240:Phorbol 12,13-Dibutyrate; D001285:Atropine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D002217:Carbachol; D011493:Protein Kinase C; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; C070515:bisindolylmaleimide I; D013755:Tetradecanoylphorbol Acetate; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one