Protein kinase C inhibitors counteract the ethanol effects on myelin basic protein expression in differentiating CG-4 oligodendrocytes.
Abnormal formation of myelin appears to be one defect contributing to the development of the neuropathology associated with the fetal alcohol syndrome. Using the CG-4 cell line we previously showed that 25-75 mM EtOH downregulates the expression of myelin basic protein (MBP) in differentiating oligodendrocytes (OLGs) without affecting morphological development (Dev. Brain Res. 128 (2001) 9). Here we showed that a relatively low concentration of 12-phorbol-13-myristate acetate (PMA) mimicked the EtOH-caused inhibition of MBP expression without affecting morphology. The inhibition of MBP expression by 100 mM EtOH or 1 nM PMA was completely counteracted by three inhibitors of protein kinase C (PKC), bisindolylmaleimide I, chelerythrine chloride, and calphostin C, indicating that EtOH downregulated MBP expression by activating PKC. We investigated whether the EtOH activation resulted, in part, from upregulation of the expression of PKC isozymes. Of 11 PKC isozymes examined, CG-4 OLGs expressed nine; PKCs alpha, beta1, beta2; delta, epsilon, eta; lambda, zeta; mu; while PKC isozymes gamma and theta were not detected. Only five PKC isozymes, alpha, beta1, beta2, eta, and mu, displayed developmental changes in expression. However, EtOH did not upregulate the early expression of any PKC isozyme during the first 2 days of differentiation, the developmental stage when it downregulates MBP expression in CG-4 cells. The similar effects of PMA and EtOH indicate that EtOH delays MBP expression by activating at least one phorbol ester-sensitive PKC isozyme in oligodendrocytes without upregulating its expression.
Bichenkov
Evgeny
E
Department of Pathology, Anatomy, and Cell Biology, 264 Jefferson Alumni Hall, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA 19107, USA.
Ellingson
John S
JS
D002454:Cell Differentiation; D002460:Cell Line; D002492:Central Nervous System Depressants / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D000431:Ethanol / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D006801:Humans; D015151:Immunoblotting; D004676:Myelin Basic Protein / Q000096:biosynthesis*; D009836:Oligodendroglia / Q000187:drug effects / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002492:Central Nervous System Depressants; D004791:Enzyme Inhibitors; D004676:Myelin Basic Protein; D000431:Ethanol; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate