GABAA Receptor-Interacting Proteins

GABAA 受体相互作用蛋白

• 批准号: 7752481
• 负责人: ANGEL L DE BLAS
• 金额: $ 35.57万
• 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/7752481
• 关键词: Affect; Antibodies; Axon; Biological Assay; Brain; Cells; Complex; Data; Development; Electron Microscopy; Epilepsy; Exocytosis; Extracellular Domain; Family; Family member; Figs - dietary; GABA-A Receptor; Glutamates; Grant; Gray unit of radiation dose; Heterogeneity; Hippocampus (Brain); Interneurons; Knowledge; Link; Mass Spectrum Analysis; Mental disorders; Molecular; NCOA2 gene; Names; Neurologic; Neurons; Play; Proteins; RNA Splicing; Rattus; Role; Signal Transduction; Small Interfering RNA; Specificity; Structure; Subcellular Fractions; Synapses; Synaptic Vesicles; Synaptic plasticity; Techniques; Testing; Time; Yeasts; density; gephyrin; hippocampal pyramidal neuron; human RIPK1 protein; immunocytochemistry; light microscopy; member; nerve supply; novel; postsynaptic; presynaptic; receptor; synaptic function; trafficking; yeast two hybrid system

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 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.

关于GABA-A受体集中在抑制性的机制的知识有限 GABA能灰色的2型突触。 GABA-A受体在大脑功能中起主要作用。它一直 估计40％的大脑突触是GABA能。该提议的长期目标是 确定与突触后聚类和锚定涉及的2型GABA能突触的蛋白质 GABA-A受体。该提议的目的是：1）证明握把蛋白参与 Gephyrin和Gaba-A受体的突触后聚类； 2）研究 原钙粘蛋白的突触聚类，其细胞内结构域与大的细胞内环相互作用 GABA-A受体的伽马2亚基和GABA-A受体的突触聚类和3） 检验以下假设：Septins 6和7，它们富含大鼠大脑中的2型突触后密度， 参与GABA-A受体聚类和/或胞吐作用。光学显微镜和电子显微镜 具有特定抗体的免疫细胞化学将用于揭示突触共定位和 完整大脑和海马培养物中GABA-A受体和突触蛋白的聚类。 突触蛋白的鉴定将通过质谱法完成。诸如siRNA处理等技术 培养的海马神经元和在这些培养物中标记的突触后蛋白的表达 完整的大脑还将用于阐明突触蛋白的突触靶向和聚类 和GABA-A受体。这些研究与理解分子成分有关 GABA能突触，突触靶向和突触后聚类以及机制 参与神经元连通性和突触可塑性。可以预测，不适当的突触和 GABA-A受体的外innaptrynaptic定位会影响GABA能突触功能和大脑发育， 导致癫痫和其他神经和精神障碍。