Chronic ethanol effects on the expression of phospholipase C isozymes and Gq/11-protein in primary cultures of astrocytes.
The goal of this investigation was to determine whether chronic ethanol exposure alters the expression of specific protein sites distal to receptors [Gq/11-protein, phospholipase C (PLC) isozymes] in primary cultures of astrocytes obtained from neonatal rat cortex. The protein expression (immunolabeling) of the PLC-beta 1, -gamma 1, -delta 1 isozymes and of the Gq/11 alpha subunit was determined by Western blot analysis using specific monoclonal antibodies. The PLC-beta 1, -gamma 1, -delta 1 isozymes and the Gq/11 alpha subunit migrated at apparent molecular masses (PLC-beta 1, 41 kDa; PLC-gamma 1, 145 kDa. PLC-delta 1, 85 kDa: Gq/11 alpha protein, 42 kDa). Thus, a PLC-beta 1 fragment of 41 kDa, but not the biologically active 150 kDa PLC-beta 1, was detected in primary cultures of astrocytes. Chronic ethanol exposure (4 days) resulted in a significant increase in the expression of PLC-delta 1, whereas under identical conditions, the expression of PLC-beta 1, -gamma 1, and of the alpha subunit of Gq/11 protein was not significantly altered in astrocytes. These results suggest that chronic ethanol exposure results in an increased expression of the PLC-delta 1, isozyme in primary cultures of astrocytes.
Department of Psychiatry, College of Medicine, University of Illinois at Chicago 60612, USA.
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Pandey
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Pandey
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Smith
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001253:Astrocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000096:biosynthesis* / Q000378:metabolism; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000096:biosynthesis*; D008970:Molecular Weight; D051381:Rats; D017207:Rats, Sprague-Dawley; D010738:Type C Phospholipases / Q000096:biosynthesis*
D000911:Antibodies, Monoclonal; D002492:Central Nervous System Depressants; D007527:Isoenzymes; D000431:Ethanol; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins