Roberts D S, Raol Y H, Bandyopadhyay S, Lund I V, Budreck E C, Passini M A, Wolfe J H, Brooks-Kayal A R, Russek S J
Laboratory of Molecular Neurobiology, Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA 02118, USA.
Proc Natl Acad Sci U S A. 2005 Aug 16;102(33):11894-9. doi: 10.1073/pnas.0501434102. Epub 2005 Aug 9.
GABA is the major inhibitory transmitter at CNS synapses. Changes in subunit composition of the pentameric GABA(A) receptor, including increased levels of alpha4 subunit in dentate granule cells and associated functional alterations such as increased zinc blockade of GABA currents, are hypothesized to be critical components of epileptogenesis. Here, we report that the minimal promoter of the human alpha4 subunit gene (GABRA4p), when used to drive reporter gene expression from adeno-associated viral vectors, controls condition-specific up-regulation in response to status epilepticus, defining a transcriptional mechanism for seizure-induced changes in levels of alpha4 subunit containing GABA(A) receptors. Transfection studies in primary hippocampal neurons show that inducible early growth response factor 3 (Egr3) up-regulates GABRA4p activity as well as the levels of endogenous alpha4 subunits. Given that Egr3 knockout mice display approximately 50% less GABRA4 mRNAs in the hippocampus and that increases in alpha4 and Egr3 mRNAs in response to pilocarpine-induced status epilepticus are accompanied by increased binding of Egr3 to GABRA4 in dentate granule cells, our findings support a role for Egr3 as a major regulator of GABRA4 in developing neurons and in epilepsy.
γ-氨基丁酸(GABA)是中枢神经系统(CNS)突触中的主要抑制性神经递质。五聚体GABA(A)受体的亚基组成变化,包括齿状颗粒细胞中α4亚基水平的增加以及相关的功能改变,如GABA电流的锌阻断增加,被认为是癫痫发生的关键组成部分。在此,我们报告,当使用人α4亚基基因的最小启动子(GABRA4p)来驱动腺相关病毒载体的报告基因表达时,它可控制对癫痫持续状态的条件特异性上调,从而确定了癫痫发作诱导含α4亚基的GABA(A)受体水平变化的转录机制。在原代海马神经元中的转染研究表明,诱导型早期生长反应因子3(Egr3)上调GABRA4p活性以及内源性α4亚基的水平。鉴于Egr3基因敲除小鼠海马中的GABRA4 mRNA减少约50%,并且在毛果芸香碱诱导的癫痫持续状态下α4和Egr3 mRNA的增加伴随着齿状颗粒细胞中Egr3与GABRA4结合的增加,我们的研究结果支持Egr3在发育中的神经元和癫痫中作为GABRA4主要调节因子的作用。