Functional Analysis of SV2 by Targeted Gene Disruption

通过靶向基因破坏对 SV2 进行功能分析

• 批准号: 6776772
• 负责人: Sandra M Bajjalieh
• 金额: $ 34.11万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6776772
• 关键词: electron microscopy; electroretinography; endocytosis; genetically modified animals; glycoproteins; green fluorescent proteins; hippocampus; laboratory mouse; molecular biology; nerve /myelin protein; neural transmission; neurons; neuroregulation; neurotransmitter transport; polymerase chain reaction; protein isoforms; protein structure function; site directed mutagenesis; synaptic vesicles; synaptotagmin; tissue /cell culture; voltage /patch clamp

DESCRIPTION (provided by applicant): Determining the molecular events that produce and regulate the release of neurotransmitters is one of the central goals of molecular neurobiology. Defining these events, and how they are altered with experience and in disease, will provide an understanding of normal processes such as learning and serve as the biological basis for developing new therapies for mental illness, neurological disorders and addiction. The work proposed here addresses the function of Synaptic Vesicle Protein 2 (SV2), a glycoprotein constituent of all neurotransmitter-containing (synaptic) vesicles. In the last funding period we demonstrated that Synaptic Vesicle Protein 2 (SV2) is an essential modulator of neurotransmission. Loss of SV2A reduces neurotransmission in hippocampal and cortical neurons, and reduces the size of the readily releasable pool of vesicles in chromaffin cells. Reduced neurotransmission is not due to morphological changes or to a decrease in morphologically "docked" synaptic vesicles. Together these results suggest that SV2 is a positive modulator of vesicle priming that acts after vesicle attachment at the plasma membrane. We propose to use SV2 knockout mice to determine SV2's molecular function in fast neurotransmission and to resolve differences in isoform functioning. Our specific aims are: 1. To determine how neurotransmission is altered in cultured hippocampal neurons from SV2 KOs. 2. To use exogeneous protein expression in hippocampal neurons to test hypotheses of SV2 function. 3. To test the hypothesis that SV2B plays a crucial role in ribbon synapse maintenance and/or functioning.

描述（由申请人提供）：确定产生和调节神经递质释放的分子事件是分子神经生物学的核心目标之一。定义这些事件以及如何通过经验和疾病改变它们，将提供对诸如学习的正常过程的理解，并成为开发新的精神疾病，神经系统疾病和成瘾疗法的生物学基础。 这里提出的工作介绍了突触囊泡蛋白2（SV2）的功能，这是所有含神经递质（突触）囊泡的糖蛋白成分的糖蛋白成分。在最后的资金期间，我们证明了突触囊泡蛋白2（SV2）是神经传递的必不可少的调节剂。 SV2A的丧失会减少海马和皮质神经元中的神经传递，并减少铬脂细胞中易于释放的囊泡池的大小。降低的神经传递不是由于形态变化或形态上“停靠”的突触囊泡的减少。这些结果共同表明，SV2是囊泡启动的正调节剂，在质膜处的囊泡附着后起作用。我们建议使用SV2基因敲除小鼠在快速神经传递中确定SV2的分子功能，并解决同工型功能的差异。我们的具体目的是： 1。确定如何在SV2 KOS培养的海马神经元中改变神经传递。 2。在海马神经元中使用外生蛋白表达来检验SV2功能的假设。 3。检验以下假设：SV2B在丝带突触维持和/或功能中起着至关重要的作用。