Mutants Showing SLO-1 Synaptic Mislocalization

显示 SLO-1 突触错位的突变体

• 批准号: 6762747
• 负责人: ZHAO-WEN WANG
• 金额: $ 16.31万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-10 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6762747
• 关键词: Caenorhabditis elegans; calcium flux; gene targeting; genetic screening; genetically modified animals; green fluorescent proteins; immunoelectron microscopy; molecular chaperones; neural transmission; neurons; neurotransmitter transport; nuclear matrix; potassium channel; protein localization; protein protein interaction; protein transport; synapses

DESCRIPTION (provided by applicant): The large-conductance calcium-activated potassium channel (BK channel, Slol) is a prominent ion channel at nerve terminals. BK channels co-localize with calcium channels at the presynaptic active zone, and serve as a potent regulator of neurotransmitter release. The function of BK channels is regulated by many factors, suggesting that the channel is potentially an important player in synaptic plasticity. Since proper subcellular localization is likely critical for BK channel function, it is important to determine how BK channel synaptic localization is achieved. Defining the mechanism of BK channel synaptic targeting and localization would be a significant advance in our understanding of synaptic structure and function. In C. elegans, loss-of-function mutations of the BK channel (SLO-1) results in increased neurotransmitter release (Wang et al., 2001), suggesting that the normal function of SLO-1 is to downregulate the release. Similar to BK channels of other species, SLO-1 is localized to synaptic regions in the nervous system (Wang et al., 2001). This restricted distribution pattern was changed to diffuse expression in neurons following a modification of the SLO-1 carboxyl terminus. This proposal seeks to test the hypothesis that presynaptic localization of BK channels is mediated by specific transport and scaffold molecules. Specifically, a genetic screen will be performed to isolate mutants showing mislocalization of a GFP-tagged SLO-1 transgene in C. elegans. Analysis of these mutants in subsequent studies may allow the identification of molecules mediating SLO-1 synaptic targeting and localization.

描述（申请人提供）：大电导钙激活钾通道（BK通道，Slol）是神经末梢的重要离子通道。 BK 通道与突触前活性区的钙通道共定位，并作为神经递质释放的有效调节器。 BK 通道的功能受多种因素调节，表明该通道可能在突触可塑性中发挥重要作用。由于正确的亚细胞定位对于 BK 通道功能可能至关重要，因此确定如何实现 BK 通道突触定位非常重要。定义 BK 通道突触靶向和定位的机制将是我们对突触结构和功能理解的重大进步。 在秀丽隐杆线虫中，BK 通道 (SLO-1) 的功能丧失突变导致神经递质释放增加（Wang 等人，2001），这表明 SLO-1 的正常功能是下调释放。与其他物种的 BK 通道类似，SLO-1 定位于神经系统的突触区域 (Wang et al., 2001)。 SLO-1 羧基末端修饰后，这种限制性分布模式变为神经元中的弥散表达。该提案旨在检验 BK 通道的突触前定位是由特定转运和支架分子介导的假设。具体来说，将进行遗传筛选以分离显示 GFP 标记的 SLO-1 转基因在秀丽隐杆线虫中错误定位的突变体。在后续研究中对这些突变体的分析可能有助于鉴定介导 SLO-1 突触靶向和定位的分子。