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

化学突触是神经元之间的接触点，因此是神经系统信息传递的中心。在突触前一侧，一个复杂的蛋白质机制介导神经递质的释放(储存在突触小泡-SVS中)，下游细胞由包含特定神经递质受体的突触后装置激活。通过纯化，我们在线虫中鉴定了与突触后烟碱型乙酰胆碱受体(NAChRs)相关的蛋白质。我们通过包括电生理学在内的方法发现，其中三种烟碱受体相关蛋白NRA-1、-2和-3调节nAChRs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，我们发现新的线虫蛋白是SVS的一部分。通过药理学、行为学和细胞生物学分析、电生理学和我们开发的一种用于外源性光刺激神经活动的新工具，光门控阳离子通道视紫红质-2(ChR2)，在(突变的)动物中对这些蛋白质进行了功能表征。具体地说，我们希望进一步开发ChR2来分析神经肌肉突触的突触传递缺陷。

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.