A role for protein kinase C and its substrates in the action of valproic acid in the brain: implications for neural plasticity.
Valproic acid (VPA) is a broad-spectrum anticonvulsant with well-documented teratogenic effects, but whose mechanism of action is largely unknown. In the present study we have examined the effects of VPA on the expression of two prominent substrates for protein kinase C (PKC) in the brain, MARCKS and GAP-43, which have been implicated in actin-membrane plasticity and neurite outgrowth during neuronal differentiation, respectively, and are essential to normal brain development. Immortalized hippocampal HN33 cells exposed to VPA exhibited reduced MARCKS protein expression and demonstrated increased GAP-43 protein expression, with concomitant alterations in cellular morphology, including an increase in the number and length of neurites and accompanied by a reduction in cell growth rate. The effects of VPA were observed at clinically relevant concentrations following chronic (>1 day) VPA exposure. We also present evidence for a VPA-induced alteration in PKC activity, as well as temporal changes in individual PKC isozyme expression. Inhibition of PKC with the PKC-selective inhibitor, LY333531, prevented the VPA-induced down-regulation of membrane-associated MARCKS, but had no effect on the cytosolic MARCKS reduction or the GAP-43 up-regulation. Inhibition of PKC by LY333531 enhanced the differentiating effects of VPA; additionally, LY333531 alone induced greater neurite outgrowth in this cell line. Collectively, these data indicate that VPA induces neuronal differentiation, associated with a reduction in MARCKS expression and an increase in GAP-43 expression, consistent with the hypothesis that a reduction in MARCKS at the membrane may be permissive for cytoskeletal plasticity during neurite outgrowth.
Watterson
Jeannette M
JM
Department of Molecular Biology and Immunology, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.
Watson
David G
DG
Meyer
Edwin M
EM
Lenox
Robert H
RH
D000818:Animals; D000927:Anticonvulsants / Q000494:pharmacology*; D001921:Brain / Q000187:drug effects* / Q000196:embryology / Q000201:enzymology; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002461:Cell Line, Transformed; D003599:Cytoskeleton / Q000187:drug effects / Q000201:enzymology; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D019922:GAP-43 Protein / Q000187:drug effects / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes / Q000187:drug effects / Q000378:metabolism; D008301:Maleimides / Q000494:pharmacology; D008565:Membrane Proteins; D051379:Mice; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D009421:Nervous System Malformations / Q000139:chemically induced / Q000201:enzymology / Q000503:physiopathology; D016501:Neurites / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D009473:Neuronal Plasticity / Q000187:drug effects* / Q000502:physiology; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology; D011247:Pregnancy; D011493:Protein Kinase C / Q000187:drug effects* / Q000378:metabolism; D011506:Proteins / Q000187:drug effects / Q000378:metabolism; D013723:Teratogens / Q000494:pharmacology; D014635:Valproic Acid / Q000494:pharmacology*
D000927:Anticonvulsants; D004791:Enzyme Inhibitors; D019922:GAP-43 Protein; D007211:Indoles; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes; C000619710:MARCKS protein, human; D008301:Maleimides; C000619711:Marcks protein, mouse; D008565:Membrane Proteins; D011506:Proteins; D013723:Teratogens; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D014635:Valproic Acid; C099154:ruboxistaurin; D011493:Protein Kinase C