Roles of Mouse Sad-1 in Presynaptic Differentiation

小鼠 Sad-1 在突触前分化中的作用

• 批准号: 6936840
• 负责人: YUCHIN Albert Pan
• 金额: $ 1.22万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6936840
• 关键词: Saccharomyces; axon; cell differentiation; cellular polarity; cytoskeleton; developmental neurobiology; fungal proteins; gene targeting; genetically modified animals; immunocytochemistry; in situ hybridization; laboratory mouse; neural transmission; neurons; phenotype; predoctoral investigator; protein localization; protein structure function; stainings; synapses; synaptic vesicles

DESCRIPTION (provided by applicant): A synapse is comprised of two compartments: presynaptic (the axon terminal) and postsynaptic (the target membrane). During the formation of a synapse, the pre- and postsynaptic compartments both differentiate. Much has been learned about postsynaptic differentiation, but the molecular machinery of presynaptic differentiation remains largely unknown. Identification of the genes that are important for presynaptic development will help fill this gap and facilitate the understanding of brain functions. In C. elegans, presynaptic differentiation requires the serine/threonine kinase Sad-1: synaptic vesicle clustering is decreased in the Sad-1 mutant and overexpression of Sad-1 causes vesicles to mislocalize. This proposed study is aimed at elucidating the functions of the murine orthologues of Sad-1 in presynaptic differentiation. There are two orthologues of Sad-1 in mice, Sad-1A and B. Double knockout mice, recently generated in our lab, are paralyzed and die at birth. The first aim is to look at Sad-1 tissue and cellular distribution by antibody staining and in situ hybridization. This will provide information about where Sad-1 is functioning. The second aim is to characterize the phenotype of the Sad-1 knockout mice. The formation of synapses as well as other aspects of neurodevelopment will be examined in tissues or isolated neurons from knockout mice. The third aim is to generate CRE mediated conditional Sad-1 knockout. This will help the animals live past birth and allow observation of postnatal synaptogenesis

描述（由申请人提供）：突触由两个隔室组成：突触前（轴突末端）和突触后（靶膜）。在突触的形成过程中，前后突触后室都有区别。关于突触后分化的知识已经很多，但是突触前分化的分子机制仍然很大程度上未知。鉴定对突触前发育很重要的基因将有助于填补这一空白并促进对大脑功能的理解。在秀丽隐杆线虫中，突触前的分化需要丝氨酸/苏氨酸激酶SAD-1：SAD-1突变体中的突触囊泡聚类减少，SAD-1的过表达导致囊泡误差。这项拟议的研究旨在阐明SAD-1在突触前分化中的鼠直系同源物的功能。在我们实验室最近产生的小鼠，SAD-1A和B的B.双敲除小鼠中，SAD-1的两名直系同源物在出生时瘫痪并死亡。第一个目的是通过抗体染色和原位杂交来研究SAD-1组织和细胞分布。这将提供有关SAD-1在哪里运行的信息。第二个目的是表征SAD-1敲除小鼠的表型。突触的形成以及神经发育的其他方面将在组织或敲除小鼠的孤立神经元中检查。第三个目的是产生CRE介导的有条件SAD-1敲除。这将有助于动物过去的出生并允许观察产后突触发生