Association of protein kinase C with GABAA receptors containing α1 and α4 subunits in the cerebral cortex:: selective effects of chronic ethanol consumption
Association of protein kinase C with GABAA receptors containing α1 and α4 subunits in the cerebral cortex:: selective effects of chronic ethanol consumption
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DOI：
10.1046/j.1471-4159.2002.00943.x
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发表时间：
2002-07-01
影响因子：
4.7
通讯作者：
Morrow, AL
中科院分区：
文献类型：
--
作者：
Kumar, S;Sieghart, W;Morrow, AL
先前的研究表明，蛋白激酶C （PKC）异构体对大脑γ -氨基丁酸(A) （GABA）受体反应的敏感性有不同的影响。PKCgamma和PKCepsilon敲除小鼠均表现出GABA(A)受体介导的Cl-通量的乙醇增强改变。此外，慢性乙醇消耗改变GABA(A)受体功能和受体亚基肽表达的机制尚不清楚。本研究探讨了PKC异构体与GABA(A)受体直接相关的可能性，这种关联受到慢性乙醇暴露的影响。利用选择性抗体从大鼠大脑皮层膜部分提取的溶解蛋白中免疫沉淀含有alpha1或alpha4亚基的GABA(A)受体。免疫沉淀受体通过western blot分析筛选PKCdelta， gamma和epsilon亚型的存在。我们发现在alpha1和alpha4亚基免疫沉淀中有PKCgamma的明显标记，而没有PKCdelta或PKCepsilon。与pkcγ抗体的免疫沉淀，而不与IgG抗体的免疫沉淀也产生了GABA(A)受体α 1和α 4亚基。长期饮用乙醇后，pkcγ与含α 1受体的关联降低了44 +/- 11%。相比之下，长期饮用乙醇后，与含有alpha4的受体相关的PKCgamma增加了32 +/- 7%。这些结果表明PKCgamma可能参与慢性乙醇消耗后GABA(A)受体的适应。
Previous studies have suggested that protein kinase C (PKC) isoforms differentially influence the sensitivity of gamma-aminobutyric acid(A) (GABA(A)) receptor responses in brain. Both PKCgamma and PKCepsilon knock-out mice exhibit altered ethanol potentiation of GABA(A) receptor mediated Cl- flux. Furthermore, chronic ethanol consumption alters GABA(A) receptor function and receptor subunit peptide expression by mechanisms that are not yet understood. The present study explored the possibility that PKC isoforms are directly associated with GABA(A) receptors, and this association is influenced by chronic ethanol exposure. GABA(A) receptors containing alpha1 or alpha4 subunits were immunoprecipitated from solubilized protein derived from the membrane fraction of rat cerebral cortex using selective antibodies. Immunoprecipitated receptors were screened by western blot analysis for the presence of PKCdelta, gamma and epsilon isoforms. We found pronounced labeling of PKCgamma but not PKCdelta or PKCepsilon in the alpha1 and alpha4 subunit immunoprecipitates. Immunoprecipitation with PKCgamma, but not with IgG antibody also yielded GABA(A) receptor alpha1 and alpha4 subunits in the immunoprecipitate. The association of PKCgamma with alpha1-containing receptors was decreased 44 +/- 11% after chronic ethanol consumption. In contrast, PKCgamma associated with alpha4-containing receptors was increased 32 +/- 7% after chronic ethanol consumption. These results suggest that PKCgamma may be involved in GABA(A) receptor adaptations following chronic ethanol consumption.