Keir W J, Morrow A L
Department of Psychiatry, University of North Carolina School of Medicine, Chapel Hill 27599.
Brain Res Mol Brain Res. 1994 Sep;25(3-4):200-8. doi: 10.1016/0169-328x(94)90154-6.
Several lines of evidence suggest an important role for ethanol interactions with GABAA receptors in the development of the ethanol withdrawal syndrome. The present study was undertaken to determine whether there is a genetic relationship between ethanol withdrawal seizure severity and the expression of particular GABAA receptor subunits in mouse lines selectively bred for differential sensitivity to ethanol withdrawal seizures. Since GABAA receptor subunit levels are subject to modulation by ethanol, the levels of GABAA receptor alpha 1, alpha 6 and beta 2 subunit mRNAs were measured in cerebellum while alpha 1 and beta 2 subunit levels were determined in cerebral cortex of ethanol-naive WSR and WSP mice. Poly(A)+ RNA was isolated from groups of 6-10 animals and the GABAA receptor subunit mRNA levels were quantified by Northern blot analysis using subunit selective cRNA probes. In the cerebellum, greater levels of each of these subunit mRNAs were detected in WSR1 mice compared to WSP1 mice. The levels of GABAA receptor alpha 1 subunit mRNAs were approximately 26 +/- 16 percent greater for the 4.4 kb transcript and 84 +/- 23 percent greater for the 4.8 kb transcript in WSR mice vs WSP mice. GABAA receptor alpha 6 subunit (2.7 kb) mRNA levels in cerebellum were 159 +/- 58 percent greater in WSR mice than WSP mice, while beta 2 subunit mRNA levels were 110 +/- 30 percent greater in WSR than WSP mice. These results were replicated for the alpha 1 and alpha 6 subunits in WSR2 vs WSP2 mouse cerebella. No differences in beta-actin mRNA levels were detected on the same RNA blots.(ABSTRACT TRUNCATED AT 250 WORDS)
多项证据表明，乙醇与γ-氨基丁酸A型（GABAA）受体的相互作用在乙醇戒断综合征的发展中起重要作用。本研究旨在确定在对乙醇戒断性癫痫具有不同敏感性而选择性培育的小鼠品系中，乙醇戒断性癫痫发作的严重程度与特定GABAA受体亚基的表达之间是否存在遗传关系。由于GABAA受体亚基水平会受到乙醇的调节，因此在未接触过乙醇的WSR和WSP小鼠的小脑中测量了GABAA受体α1、α6和β2亚基mRNA的水平，同时在大脑皮层中测定了α1和β2亚基的水平。从6 - 10只动物的群体中分离出多聚腺苷酸（Poly(A)+）RNA，并使用亚基选择性cRNA探针通过Northern印迹分析对GABAA受体亚基mRNA水平进行定量。在小脑中，与WSP1小鼠相比，WSR1小鼠中这些亚基mRNA的水平更高。与WSP小鼠相比，WSR小鼠中4.4 kb转录本的GABAA受体α1亚基mRNA水平大约高26±16%，4.8 kb转录本高84±23%。小脑中GABAA受体α6亚基（2.7 kb）的mRNA水平在WSR小鼠中比WSP小鼠高159±58%，而β2亚基mRNA水平在WSR小鼠中比WSP小鼠高110±30%。在WSR2和WSP2小鼠的小脑中，α1和α6亚基也得到了同样的结果。在相同的RNA印迹上未检测到β-肌动蛋白mRNA水平的差异。（摘要截断于250字）
