Brain Myosin5 and Synaptic Plasticity

脑肌球蛋白5和突触可塑性项目摘要人类的大脑是一个灵活的结构，它会随着经验的变化而变化，但人们并不完全理解经验是如何在细胞水平上改变大脑并导致学习和记忆的。这个提议提出的问题是“肌动蛋白依赖的分子马达，脑肌球蛋白5，在经验驱动的突触可塑性中扮演什么角色，即肌球蛋白5如何参与突触强化？“脑肌球蛋白5（Myosin-Va）具有多种功能，将其与突触可塑性联系起来。它在轴突终末和树突棘上运输囊泡以及其他种类的货物。以往的研究表明，myosin 5与驱动蛋白（heteromotor complex）结合，是内质网（endoplasmic reticulum，ER）小泡转运的分子马达。这些研究引出了本项目要解决的具体问题：肌球蛋白5和驱动蛋白如何协调ER囊泡的转运。具体目标如下：PI和他的研究小组将使用基于亲和性的生化方法分离myosin 5的脑特异性结合伴侣，并使用蛋白质组学方法（脑特异性myosin 5蛋白质组）鉴定蛋白质。蛋白质组学筛选的结果将用于确定哪些蛋白质构成肌球蛋白5/驱动蛋白异源运动复合体。从蛋白质组学鉴定的蛋白质列表中，他们将鉴定与肌球蛋白5/驱动蛋白运动复合体相关的信号蛋白，并使用体外运动分析确定这些信号蛋白中的哪一种在ER囊泡转运过程中起协调驱动蛋白和肌球蛋白5活性的作用。更广泛的影响将包括通过达特茅斯的欧内斯特埃弗雷特正义计划，由PI指导少数民族学生，资助本科生在他的实验室从事独立研究项目和荣誉论文研究，并与来自斯佩尔曼学院和莫尔豪斯学院（2 HBCU）的学生一起工作，他们作为交换生来到达特茅斯。此外，PI将继续在他的实验室在海洋生物实验室，伍兹霍尔MA的本科生。每年夏天，他都会就他的研究发表演讲，并为MBL的SPINES（神经）课程的少数研究生提供指导。

Brain Myosin5 and synaptic plasticityAbstract of projectThe human brain is a flexible structure that changes in response to experiences but it isnot fully understood how experience changes the brain at the cellular level and resultsin learning and memory. The question posed in this proposal is "what is the role ofthe actin-dependent molecular motor, brain myosin5, in experience-driven synapticplasticity, i.e how does myosin5 participate in synaptic strengthening?" Brainmyosin5 (Myosin-Va) has multiple functions that link it to synaptic plasticity. Ittransports vesicles as well as other kinds of cargo at axon terminals and dendriticspines. In addition, it is a constitutive component of the postsynaptic density.Previous studies have shown that myosin5 binds to kinesin (heteromotor complex)and functions as a molecular motor for the transport of endoplasmic reticulum (ER)vesicles. Those studies led to the following specific question to be addressed in thisproject: How is transport of ER vesicles by myosin5 and kinesin coordinated. Thespecific aims are as follows: The PI and his research group will isolate brain-specificbinding partners of myosin5 using affinity-based biochemical methods and identifythe proteins using proteomic approaches (brain-specific myosin5 proteome). Theresults from this proteomics screen will be used to determine which proteinsconstitute the myosin5/kinesin heteromotor complex. From the list of proteinsidentified by proteomics, they will identify signaling proteins that associate with themyosin5/kinesin motor complex and determine which of these signaling proteinsfunction to coordinate kinesin and myosin5 activity during transport of ER vesiclesusing an in vitro motility assay.Broader impacts resulting from the proposed activities: The broader impacts willinclude mentoring by the PI of minority students through the Ernest Everett JustProgram at Dartmouth, sponsoring undergraduate students engaged in independentresearch projects and honor's theses research in his laboratory and working withstudents from Spelman College and Morehouse College (2 HBCUs) who come toDartmouth as exchange students. In addition, the PI will continue to sponsorundergraduate students in his laboratory at the Marine Biological Laboratory, WoodsHole MA. Each summer he gives lectures on his research and provides mentoring tominority graduate students enrolled in the SPINES (Neuro) course at the MBL.