Analysis of the GABA-A receptor alpha-4 promoter

描述（由申请人提供）：本研究项目的目的是 研究GABAA受体α 4亚单位表达的调控 （GABAA-R），它显示出一定程度的可塑性，这是显着的， GABAA-R亚基家族，在治疗期间表达显著增加。 各种过度兴奋综合征，如慢性戒断综合征， 间歇性乙醇（CIE）暴露。导致这种情况的遗传因素 将研究小鼠GABAA-Ra 4亚基表达的控制。的 本发明的具体目的是：1）克隆编码 小鼠GABAA-Ra 4亚基（GABRA 4），包括推定的5 ′-调节结构域， 并表征基因结构和外显子/内含子边界。2)到 研究mRNA编码的转录起始位点， GABAA-Ra 4亚基。3)为了调查和鉴定假定的消音器， 小鼠中的增强子、引发子和/或位置调节元件 α 4亚基基因。α 4亚基的表达仅限于特定的 在非神经元细胞中不存在-这可能是由于 一个或多个神经元限制性沉默元件的存在， 基因的5 '-调控结构域。4)开发一个细胞系或原代神经元 培养系统以研究GABAA-Ra 4亚基的表达调控， 这将概括该基因在体内诱导的关键特征， 过度兴奋的模型反复服用酒精， 原代培养的神经元，或细胞系模型，导致诱导 a4亚基的表达，并且这种表达的增加可以是 通过抑制特定的信号通路来调节， 转录调控这些目标的实现将提高我们的 GABAA-R亚单位的知识与行为相关 超兴奋性，并显示出相当大的可塑性。这些知识将 在涉及诱导的转录机制的研究中是有用的 GABAA-R亚基表达，并可能在设计干预措施， 中断与戒断过度兴奋相关的基因组机制。

DESCRIPTION (provided by applicant): The objective of this research project is to study the regulation of expression of the a4 subunit of the GABAA receptor (GABAA-R), which shows a degree of plasticity that is remarkable among the family of GABAA-R subunits, with a striking increase in expression during a variety of hyper-excitability syndromes, such as withdrawal from chronic intermittent ethanol (CIE) exposure. The genetic elements responsible for the control of expression of the mouse GABAA-R a4 subunit will be studied. The specific aims of the present proposal are: 1) To clone the gene encoding the mouse GABAA-R a4 subunit (GABRA4), including the putative 5'-regulatory domain, and to characterize the gene structure and exon/intron boundaries. 2) To investigate the transcriptional start sites for mRNA species encoding the GABAA-R a4 subunit. 3) To investigate and characterize putative silencer, enhancer, initiator and/or positional regulator element/s in the mouse a4-subunit gene. Expression of the a4 subunit is limited to specific populations of neurons and is absent in non-neuronal cells - this may be due to the existence of one or more neuron-restrictive silencer elements in the 5'-regulatory domain of the gene. 4) To develop a cell line or primary neuronal culture system to study the regulation of expression of the GABAA-R a4 subunit, that will recapitulate key features of the induction of this gene in vivo in models of hyperexcitability. Repeated administration of alcohol, either to neurons in primary culture, or to a cell line model, results in induction of expression of the a4 subunit, and that this increased expression can be regulated by the inhibition of specific signaling pathways associated with transcriptional regulation. The completion of these Aims will increase our knowledge of a GABAA-R subunit that is associated with behavioral hyperexcitability and shows substantial plasticity. This knowledge will be useful in the study of transcriptional mechanisms involved in the induction of GABAA-R subunit expression, and potentially in designing interventions aimed at interrupting genomic mechanisms associated with withdrawal hyperexcitability.