GABAA Receptor-Interacting Proteins

GABAA 受体相互作用蛋白

• 批准号: 7139770
• 负责人: ANGEL L DE BLAS
• 金额: $ 22.2万
• 依托单位: UNIVERSITY OF CONNECTICUT STORRS
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7139770
• 关键词: GABA receptor; confocal scanning microscopy; guanine nucleotide exchange factors; protein localization; protein protein interaction; protein sequence; protein structure function; receptor binding; synapses; yeast two hybrid system

DESCRIPTION (provided by applicant): There is limited knowledge about the mechanisms by which GABA-A receptors are concentrated at inhibitory GABAergic Gray's type-2 synapses. The GABA-A receptors play a major role in brain function. It has been estimated that 40% of the brain synapses are GABAergic. The long-term objective of this proposal is to identify the proteins of the type-2 GABAergic synapses involved in the postsynaptic clustering and anchoring of GABA-A receptors. The aims of this proposal are: 1) To demonstrate that GRIP proteins are involved in the postsynaptic clustering of gephyrin and GABA-A receptors; 2) To study the relationship between the synaptic clustering of a protocadherin, whose intracellular domain interacts with the large intracellular loop of the gamma 2 subunit of the GABA-A receptor, and the synaptic clustering of GABA-A receptors and 3) To test the hypothesis that septins 6 and 7, which are enriched in type-2 postsynaptic densities in the rat brain, are involved in GABA-A receptor clustering and/or exocytosis. Light microscopy and electron microscopy immunocytochemistry with specific antibodies will be used for revealing the synaptic co-localization and clustering of GABA-A receptors and synaptic proteins in the intact brain and in hippocampal cultures. Identification of synaptic proteins will be done by mass spectrometry. Techniques such as siRNA treatment of cultured hippocampal neurons and the expression of tagged postsynaptic proteins in these cultures and in the intact brain will also be used to elucidate the synaptic targeting and clustering of the synaptic proteins and GABA-A receptors. These studies are relevant for understanding the molecular components of GABAergic synapses, their synaptic targeting and postsynaptic clustering as well as the mechanisms involved in neuronal connectivity and synaptic plasticity. It is predicted that the inappropriate synaptic and extrasynaptic localization of GABA-A receptors affects GABAergic synaptic function and brain development, leading to epilepsy and other neurological and mental disorders.

描述（由申请人提供）：关于GABA-A受体集中在抑制性Gabaergic Gray 2型突触的机制的知识有限。 GABA-A受体在大脑功能中起主要作用。据估计，40％的大脑突触是GABA能。该提案的长期目标是确定与GABA-A受体的突触后聚类和锚定有关的2型GABA能突触的蛋白质。该提案的目的是：1）证明抓地力蛋白参与gephyrin和Gaba-A受体的突触后聚类； 2）研究原粘蛋白的突触聚类之间的关系，其细胞内结构域与GABA-A受体的伽马2亚基的较大的细胞内环与GABA-A受体的突触聚类与GABA-A受体的突触聚类相互作用，而GABA-A受体的突触聚类和3）与SEPTINS 6和7相互作用，该假说与脑部6和7相关。 GABA-A受体聚类和/或胞吐作用。光学显微镜和电子显微镜免疫细胞化学具有特定抗体将用于揭示完整大脑和海马培养物中GABA-A受体和突触蛋白的突触共定位和聚类。突触蛋白的鉴定将通过质谱法完成。诸如培养的海马神经元的siRNA处理以及在这些培养物中和完整大脑中标记的突触后蛋白的表达等技术也将用于阐明突触蛋白和GABA-A受体的突触靶向和聚类。这些研究与理解GABA能突触的分子成分有关，它们的突触靶向和突触后聚类以及涉及神经元连通性和突触可塑性的机制。据预测，GABA-A受体的不当突触和突触外定位会影响GABA能突触功能和大脑发育，从而导致癫痫以及其他神经系统和精神疾病。