CAM-1 function at the C. elegans neuromuscular junction

描述(由申请人提供)： 许多滥用和治疗药物的作用是通过改变突触传递的效果来实现的。同样，许多神经疾病的病理生理学根源是突触功能障碍。然而，调节突触结构的组织和维持的过程在很大程度上仍然是未知的。受体酪氨酸激酶(RTK)调节突触后神经递质受体的聚集和突触突起的生长。线虫RTK CAM-1与哺乳动物神经肌肉接头(NMJ)突触后乙酰胆碱受体(AChRs)聚集所需的肌肉特异性激酶(Musk)和哺乳动物RTK家族中的Ror家族相关。ROR RTK在不同物种中高度保守，在哺乳动物神经系统中广泛表达。然而，Ror RTK在神经系统中的功能作用尚不清楚。我们的初步结果表明，CAM-1定位于线虫NMJ，并且cam-1基因的靶向突变导致线虫神经肌肉突触的突触传递改变。本方案中的实验旨在测试CAM-1在线虫NMJ中的功能作用。我们将确定CAM-1的亚细胞分布，并评估它是否与特定的神经递质受体共定位。此外，我们将确定CAM-1是否在物理上与参与神经传递的特定突触蛋白相互作用。最后，为了确定Ror RTK的潜在配体和下游效应因子，我们将筛选线虫肌肉中过表达CAM-1引起的瘫痪的抑制因子。这项研究计划将对CAM-1和ROR激酶的潜在突触功能提供新的见解。拟议的实验还将为申请者提供线虫研究技术方面的宝贵培训。通过包括正式讲座、非正式陈述和研究在内的多样化培训计划，申请者将在线虫分子遗传方法的设计和实施方面获得新的经验。这些方法将补充申请者之前在离子通道生理学方面的培训，最终使申请者能够指导专注于线虫突触功能的生物调节的独立研究计划。Maricq实验室将基因技术与电生理方法相结合，使这里成为拟议培训的理想地点。

DESCRIPTION (provided by applicant): A number of drugs of abuse and therapeutics act by altering the efficacy of synaptic transmission. Likewise, the pathophysiology of many neurological disorders is rooted in synaptic dysfunction. However, the processes that regulate the organization and maintenance of the synaptic architecture remain largely unknown. Receptor tyrosine kinases (RTKs) have been shown to regulate neurite outgrowth and the clustering of post-synaptic neurotransmitter receptors. The C. elegans RTK CAM-1 is related to both Muscle-specific kinase (MUSK), an RTK required for clustering of post-synaptic acetylcholine receptors (AChRs) at the mammalian neuromuscular junction (NMJ), and the mammalian Ror family of RTKs. Ror RTKs are highly conserved across species and are widely expressed in the mammalian nervous system. However, the functional roles of Ror RTKs in the nervous system are unknown. Our preliminary results show that CAM-1 is localized to the C. elegans NMJ and that targeted mutation of the cam-1 gene results in altered synaptic transmission at C. elegans neuromuscular synapses. The experiments in this proposal are designed to test the functional role of CAM-1 at the C. elegans NMJ. We will determine the subcellular distribution of CAM-1 and assess whether it is colocalized with specific neurotransmitter receptors. Furthermore, we will determine if CAM-1 physically interacts with specific synaptic proteins involved in neurotransmission. Finally, to identify potential ligands and downstream effectors for Ror RTKS, we will screen for suppressors of the paralysis caused by overexpression of CAM-1 in C. elegans muscles. This research program will provide novel insights into potential synaptic functions for CAM-1 and Ror kinases in general. The proposed experiments will also provide valuable training for the applicant in C. elegans research techniques. Through a diverse training program including formal lectures, informal presentations and research, the applicant will gain new experience in the design and implementation of C. elegans molecular genetic approaches. These approaches will complement the applicant's previous training in ion channel physiology, ultimately enabling the applicant to direct an independent research program focused upon the biological regulation of synaptic function in C. elegans. The application of genetic techniques in concert with electrophysiological approaches in the Maricq laboratory makes this an ideal location for the proposed training.