CAM-1 function at the C. elegans neuromuscular junction

CAM-1 在秀丽隐杆线虫神经肌肉接头处的功能

• 批准号: 7091652
• 负责人: MICHAEL M FRANCIS
• 金额: $ 2.3万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7091652
• 关键词: Caenorhabditis elegans; biological signal transduction; electrophysiology; ligands; molecular genetics; neuromuscular junction; neurotransmitter receptor; protein localization; protein structure function; protein tyrosine kinase; receptor expression

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.

描述（由申请人提供）： 许多滥用药物和治疗药物通过改变突触传递的功效起作用。同样，许多神经系统疾病的病理生理学根源于突触功能障碍。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未知。受体酪氨酸激酶（RTK）已被证明可以调节神经突起的生长和突触后神经递质受体的聚集。梭线虫RTKCAM-1与肌肉特异性激酶（Muscle-specific kinase，MUSK）（一种在哺乳动物神经肌肉接头（NMJ）处聚集突触后乙酰胆碱受体（AChR）所需的RTK）和RTK的哺乳动物Ror家族相关。Ror RTK在物种间高度保守，并且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C。并且cam-1基因的靶向突变导致C.神经肌肉突触。本实验旨在检测CAM-1在C.秀丽隐杆线虫NMJ.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是否与参与神经传递的特定突触蛋白发生物理相互作用。最后，为了鉴定Ror RTKS的潜在配体和下游效应物，我们将筛选C.优雅的肌肉这项研究计划将为CAM-1和Ror激酶的潜在突触功能提供新的见解。所提出的实验也将为申请人在C. elegans研究技术。通过一个多样化的培训项目，包括正式的讲座，非正式的演讲和研究，申请人将获得新的经验，在设计和实施C。elegans分子遗传学方法这些方法将补充申请人之前在离子通道生理学方面的培训，最终使申请人能够指导专注于C中突触功能的生物调节的独立研究计划。优美的在Maricq实验室中，遗传技术与电生理方法的应用使其成为拟议培训的理想场所。