Postsynaptic Rapsyn Dynamics at the Neuromuscular Junction of Living Animals

活体动物神经肌肉接头处的突触后 Rapsyn 动力学

• 批准号: 0718211
• 负责人: Mohammed Akaaboune
• 金额: $ 37.5万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0718211&HistoricalAwards=false
• 关键词: Postsynaptic; Rapsyn; Dynamics; Neuromuscular; Junction

The efficiency of nerve impulse transmission to muscle cells depends on the number and density of acetylcholine receptor proteins on the muscle membrane. These receptors are localized to specialized structures known as synapses, characterized by folds in the postsynaptic membrane. Receptors are firmly held in the crest of the folds by a host of specialized scaffold proteins. The goal of this study is to investigate the dynamics of one critical component of this scaffold, rapsyn, when levels of synaptic activity are manipulated. To address the dynamics of rapsyn at the synapse, the PI will fuse jellyfish green fluorescent protein (GFP) to rapsyn and introduce the construct into the neck of the mouse (sternomastoid muscle). This makes it possible to monitor and image a particular synapse over the course of several days in the living animal. Receptor proteins will be labeled by fluorescent bungarotoxin, a snake venom that binds with high affinity to the receptor. The PI will investigate the following questions: first, what is the lifetime of rapsyn at individual synapse in vivo. Second what is the effect of rapsyn overexpression on the metabolic stability of receptors in vivo. Finally, the PI will investigate the effect of synaptic activity on the dynamics of rapsyn at individual synapses in vivo. The PI will promote the training of graduate, undergraduate and high school students, including those of underrepresented groups. The PI has extensive experience in training undergraduate students. In addition this proposal will enhance teaching performance and learning in the areas of neuroscience. Results will be disseminated to the scientific community through publications and by presenting findings in meetings. Finally the PI's lab will be involved in outreach to the community.

神经冲动传递到肌肉细胞的效率取决于肌肉膜上乙酰胆碱受体蛋白的数量和密度。这些受体定位于称为突触的专门结构，其特征在于突触后膜中的褶皱。受体被一系列特殊的支架蛋白牢固地固定在褶皱的顶部。本研究的目的是研究当操纵突触活动水平时，这种支架的一个关键成分rapsyn的动力学。 为了解决rapsyn在突触处的动力学，PI将水母绿色荧光蛋白（GFP）融合到rapsyn，并将构建体引入小鼠颈部（胸乳肌）。这使得有可能在活体动物的几天内监测和成像特定的突触。受体蛋白将被荧光银环蛇毒素标记，这是一种与受体结合的高亲和力蛇毒。PI将研究以下问题：首先，rapsyn在体内单个突触的寿命是多少。第二，rapsyn过表达对受体体内代谢稳定性的影响。最后，PI将研究突触活动对体内单个突触中rapsyn动力学的影响。PI将促进对研究生、本科生和高中生的培训，包括那些代表性不足的群体。PI在培养本科生方面拥有丰富的经验。此外，该提案将提高神经科学领域的教学效果和学习效果。将通过出版物和在会议上介绍调查结果向科学界传播结果。最后，PI的实验室将参与社区外展活动。