RIG: Postsynaptic Molecular Assembly at the Onset of Neuromuscular Synaptogenesis in vivo

RIG：体内神经肌肉突触发生开始时的突触后分子组装

• 批准号: 0818243
• 负责人: Franklin Carrero-Martinez
• 金额: $ 17.42万
• 依托单位: University of Puerto Rico Mayaguez
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2011-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0818243&HistoricalAwards=false
• 关键词: RIG; Postsynaptic; Molecular; Assembly; Onset

The mechanisms that initiate a synapse have rarely been analyzed in living cells due to limited access in embryos. The investigator has previously observed filopodial processes (myopodia) extending from muscle cells toward neurons prior to neuromuscular synaptogenesis, using three- and four-dimensional imaging of fluorescent-labeled bioprobes in live Drosophila embryos. This appearance of myopodia was seen to coincide with motoneuron outgrowth, suggesting a mechanism in which synaptic partners interact before they make permanent contact. As development continues, transient stabilization of pre- and post-synaptic filopodia from matched partners can be observed. This myopodial clustering event is the focus of the current project. The investigator will test the "synaptic nucleation" hypothesis, which states that myopodial clustering serves as a sub cellular signaling compartment where the nascent synapse is coordinated. Using in vivo imaging and immunocytochemistry, the researcher will study the underlying molecules involved in synaptic partner-induced myopodial clustering, and in particular, will investigate how these proteins are recruited to the myopodial cluster compartment. The goal of this project is to gain a better understanding of synaptic matchmaking and synaptogenesis by identifying the postsynaptic molecular players at the site of synapse formation and to characterize the mechanism by which they are recruited. Once considered as passive players, muscle cells are emerging as active synaptogenic partners, and myopodial interactions offer a chance to dissect the molecular integrations that lead to the successful formation of the synapse. With over 99% of the total undergraduate student enrollment in the Department of Biology at the University of Puerto Rico Mayaguez identified as members of underrepresented groups in the sciences, this proposal will support training of a significant number of minority students. Their research experiences will be enhanced through access to mentors and role models which will be facilitated by the investigator.

启动突触的机制很少在活细胞中进行分析，因为在胚胎中的访问有限。研究人员先前观察到丝状伪足过程（肌足）从肌肉细胞向神经元延伸神经肌肉突触发生之前，使用三维和四维成像的荧光标记的生物探针在活果蝇胚胎。这种肌足的出现与运动神经元的生长相一致，表明突触伴侣在永久接触之前相互作用的机制。随着发育的继续，可以观察到来自匹配伴侣的突触前和突触后丝状伪足的短暂稳定。这种肌足群集事件是当前项目的焦点。研究者将测试“突触成核”假说，该假说指出肌足群集作为一个亚细胞信号室，新生突触在此协调。使用体内成像和免疫细胞化学，研究人员将研究参与突触伴侣诱导的肌足群集的潜在分子，特别是将研究这些蛋白质如何被招募到肌足群集隔室。该项目的目标是通过识别突触形成部位的突触后分子参与者并描述其招募机制来更好地理解突触配对和突触发生。一旦被认为是被动的球员，肌肉细胞正在成为积极的突触伙伴，和肌足的相互作用提供了一个机会，解剖的分子整合，导致突触的成功形成。超过99%的本科生在生物系在波多黎各马亚圭斯大学入学确定为科学代表性不足的群体的成员，这项建议将支持大量的少数民族学生的培训。他们的研究经验将通过获得导师和榜样，这将是由调查员促进增强。