Brain Myosin5 and Synaptic Plasticity

脑肌球蛋白5和突触可塑性项目摘要人脑是一种灵活的结构，它会随着经验的变化而变化，但人们并不完全了解经验是如何在细胞水平上改变大脑以及学习和记忆的结果。这个提案提出的问题是“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如何参与突触加强？”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上运输囊泡和其他类型的货物。此外，它还是突触后密度的组成部分。以往的研究表明，肌球蛋白5与动蛋白(异位运动复合体)结合，并作为内质网小泡运输的分子马达发挥作用。这些研究导致了本项目要解决的以下具体问题：肌球蛋白5和动蛋白是如何协调内质网小泡的运输的。具体目标如下：PI和他的研究小组将使用基于亲和力的生物化学方法分离大脑特异的肌球蛋白5结合伙伴，并使用蛋白质组学方法(大脑特异的肌球蛋白5蛋白质组)鉴定蛋白质。这个蛋白质组学筛选的结果将被用来确定哪些蛋白质构成了肌球蛋白5/动蛋白异构体复合体。从蛋白质组学鉴定的蛋白质清单中，他们将识别与肌球蛋白5/肌球蛋白运动复合体相关的信号蛋白，并使用体外动力分析确定这些信号蛋白中哪些起到协调肌动蛋白和肌球蛋白5活性的作用。拟议活动产生的广泛影响包括由达特茅斯大学少数族裔学生的PI通过欧内斯特·埃弗雷特·贾斯特项目进行指导，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，以及与来自斯皮尔曼学院和莫尔豪斯学院(2HBCU)的学生作为交换生来到达特茅斯大学。此外，PI将继续资助他在马萨诸塞州伍兹霍尔海洋生物实验室的实验室的本科生。每年夏天，他都会讲授他的研究，并为在MBL注册的脊椎(神经)课程的少数专业研究生提供指导。

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.