Protein kinase C activators work in synergy with specific growth factors to initiate tyrosine hydroxylase expression in striatal neurons in culture.
Our previous studies indicate that, in the noncatecholamine (non-CA) neurons of the striatum, expression of the gene for the CA biosynthetic enzyme tyrosine hydroxylase (TH) can be initiated by the synergistic interaction of acidic fibroblast growth factor (aFGF) and a second partner molecule. In this study, we sought to determine whether the activators of protein kinase C (PKC) signaling pathways, either alone or in conjunction with various growth factors, is sufficient to induce TH in striatal neurons. We found that when the active beta from of 4 beta-12-O-tetradecanoylphorbol 13-acetate (TPA), but not the inactive alpha analogue, was incubated in the presence of aFGF, basic FGF, or brain-derived neurotrophic factor, TH expression was initiated. Activation of the PKC pathways alone (in the absence of growth factors) did not mimic these effects, suggesting that multiple pathway activation is required for novel TH expression. Although other specific activators of PKC were effective growth factor partners, TPA was the most potent with an ED50 of 0.008 muM. Conversely, inhibitors of protein kinases, such as H7, H8, or H89, prevented the expression of TH by aFGF and TPA. Because pretreatment with protein (cycloheximide) or RNA synthesis (amanitin and actinomycin D) inhibitors eliminated the inductive effect of aFGF and TPA, we conclude that de novo transcription and translation are necessary for the expression of TH after convergence of both PKC and growth factor pathways.
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Department of Neurobiology and Anatomy, Medical College of Pennsylvania, Philadelphia, USA.
Iacovitti
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D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology; D002478:Cells, Cultured / Q000187:drug effects / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006133:Growth Substances / Q000494:pharmacology*; D051379:Mice; D017072:Neostriatum / Q000166:cytology; D009474:Neurons / Q000187:drug effects / Q000201:enzymology*; D010641:Phenotype; D011247:Pregnancy; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013755:Tetradecanoylphorbol Acetate / Q000031:analogs & derivatives / Q000494:pharmacology; D013997:Time Factors; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics / Q000378:metabolism*
D019208:Brain-Derived Neurotrophic Factor; D002273:Carcinogens; D004791:Enzyme Inhibitors; D006133:Growth Substances; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D014446:Tyrosine 3-Monooxygenase; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate