Functional analysis of novel proteins associated with nicotinic acetylcholine receptors and synaptic vesicles in Caenorhabditis elegans

与秀丽隐杆线虫烟碱乙酰胆碱受体和突触小泡相关的新型蛋白质的功能分析

• 批准号: 46383571
• 负责人: Professor Dr. Alexander Gottschalk
• 金额: --
• 依托单位: Buchmann Institute for Molecular Life Sciences
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/46383571?language=en
• 关键词: Functional; analysis; novel; proteins; associated

Chemical synapses are contact sites between neurons and thus central for information transfer in the nervous system. At the pre-synaptic side, a complex protein machinery mediates release of neurotransmitters (stored in synaptic vesicles - SVs), and the downstream cell is activated by a post-synaptic apparatus containing specific neurotransmitter receptors. By purification, we identified proteins associated with post-synaptic nicotinic acetylcholine receptors (nAChRs) in the nematode Caenorhabditis elegans. As we showed by methods including electrophysiology, three of these nicotinic receptor associated proteins, NRA-1, -2 and -3, regulate synaptic density or functional properties of nAChRs. Function of these proteins shall now be characterized in detail. Similarly, via purification and cross-species conservation, we identify novel C. elegans proteins as part of SVs. These proteins are functionally characterized in (mutant) animals lacking them, by pharmacological, behavioural and cell biological assays, electrophysiology, and a new tool we developed for exogenous photo-stimulation of neural activity, the light-gated cation channel Channelrhodopsin-2 (ChR2). Specifically, we want to further develop ChR2 for the analysis of defects in synaptic transmission at the neuromuscular synapse.

化学突触是神经元之间的接触点，因此是神经系统中信息传递的中心。在突触前侧，复杂的蛋白质机制介导神经递质（储存在突触囊泡-SV中）的释放，下游细胞被含有特定神经递质受体的突触后装置激活。通过纯化，我们确定了与突触后烟碱乙酰胆碱受体（nAChRs）在线虫秀丽隐杆线虫。正如我们通过包括电生理学在内的方法所示，这些烟碱受体相关蛋白中的三种NRA-1、NRA-2和NRA-3调节nAChR的突触密度或功能特性。现在将详细描述这些蛋白质的功能。同样，通过纯化和跨物种保护，我们确定了新的C。elegans蛋白作为SV的一部分。这些蛋白质的功能特点是在（突变）动物缺乏它们，药理学，行为和细胞生物学测定，电生理学，和一个新的工具，我们开发的外源性光刺激的神经活动，光门控阳离子通道视紫红质-2（ChR 2）。具体来说，我们希望进一步开发ChR 2用于分析神经肌肉突触的突触传递缺陷。