The Role of Kinesin Motors in Actively Delivering Cargos into Dendritic Spines

驱动蛋白马达在主动将货物输送到树突棘中的作用

基本信息

批准号：
9103889
负责人：
Derrick Mcvicker
金额：
$ 2.96万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2016-12-02
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9103889
关键词：
Actomyosin Affect Alzheimer&apos s Disease Brain Brain-Derived Neurotrophic Factor Cells Complement Confocal Microscopy Cytoskeletal Proteins Defect Dendritic Spines Development Diffusion Disease Event F-Actin FMR1 Family member Fluorescence Microscopy Fragile X Syndrome Goals Head Hippocampus (Brain)Human Individual Intellectual functioning disability Invaded Kinesin Label Lead Learning Life Lipids Maintenance Mediating Memory Microtubule Polymerization Microtubules Molecular Molecular Motors Morphology Motor N-Methyl-D-Aspartate Receptors Nervous system structure Neurodegenerative Disorders Neurons Organism Patients Phenotype Physiological Physiology Play Process Proteins Rattus Reagent Role Route Structure Synapses Synaptic plasticity Techniques Testing Tubulin Vertebral column base density excitatory neuron immunocytochemistry induced pluripotent stem cell insight knock-down microtubule-associated protein 1B novel overexpression postsynaptic postsynaptic neurons presynaptic density protein 95 public health relevance small hairpin RNA synaptotagmin IV trafficking

项目摘要

DESCRIPTION (provided by applicant): The long term goals of this project are to elucidate a novel mechanism by which microtubule (MT) invasions of dendritic spines provide a direct route for the transport of postsynaptic material into spines via MT-based molecular motor proteins. In doing so we will identify the specific motor proteins and their cargos which are involved in this process. Additionally, we will investigate the role kinesin motors and MTs play in fragile X syndrome (FXS), a disease that affects spine morphology and composition, resulting in intellectual and developmental disabilities. We have previously demonstrated that MT invasions into spines increase the amount of postsynaptic density-95 (PSD- 95) protein and that spines enlarge when invaded by MTs in an NMDA receptor-dependent manner. However, to date, there is no direct mechanism which demonstrates the precise role MTs play in synaptic plasticity. We hypothesize that the molecular motor protein kinesin can directly move cargos into spines during MT invasions and is essential for the transport of postsynaptic material critical for the formation and plasticity of mature functional synapses. In Aim 1, we will test the hypothesis that kinesin motors directly enter spines via MT invasions and determine which kinesin family members are involved in this process. We will use multi-wavelength live-cell total internal fluorescence microscopy (TIRFM) to visualize fluorescently tagged kinesin-1, 2 and 3 family members and fluorescently labeled tubulin and/or EB3 in mature rat hippocampal neurons. In Aim 2 we will test the hypothesis that specific kinesin family members associate with and transport unique cargos into dendritic spines in an activity based manner. We will test this hypothesis using a variety of techniques to visualize both fluorescently-labeled kinesin family members and cargos, and determine which kinesin family members are responsible for transporting specific cargos into spines prior to and after bouts of synaptic activity (e.g. by BDNF-induced plasticity or activation of NMDA receptors). In Aim 3 we will test the hypothesis that neurons derived from human induced pluripotent stem cells (hiPSC) from FXS patients have defects in MT and kinesin invasions into dendritic spines. Together these studies will provide fundamental insights into a novel mechanism underlying synaptic plasticity and may have implications for a devastating developmental disease.

描述（由申请人提供）：该项目的长期目标是阐明一种新机制，通过该机制，微管（MT）侵入树突棘，为通过基于 MT 的分子运动蛋白将突触后物质转运到树突棘中提供直接途径。在此过程中，我们将鉴定参与该过程的特定运动蛋白及其货物，此外，我们将研究驱动蛋白马达和 MT 在脆性 X 综合征中的作用。 (FXS)，一种影响脊柱形态和组成的疾病，导致智力和发育障碍。我们之前已经证明，MT 侵入脊柱会增加突触后密度 95 (PSD-95) 蛋白的数量，并且当 MT 侵入脊柱时，脊柱会增大。然而，迄今为止，还没有直接的机制证明 MT 在突触可塑性中发挥的精确作用。我们发现分子运动蛋白驱动蛋白可以直接移动货物。在 MT 入侵期间进入脊柱，对于突触后物质的运输至关重要在目标 1 中，我们将测试驱动蛋白马达通过 MT 侵入直接进入棘突的假设，并确定哪些驱动蛋白家族成员参与此过程。荧光显微镜 (TIRFM) 可可视化成熟大鼠中荧光标记的驱动蛋白 1、2 和 3 家族成员以及荧光标记的微管蛋白和/或 EB3在目标 2 中，我们将测试特定驱动蛋白家族成员以基于活动的方式与树突棘相关联并将独特的货物运输到树突棘中的假设，我们将使用各种技术来测试该假设，以可视化荧光标记的驱动蛋白家族成员。和货物，并确定哪些驱动蛋白家族成员负责在突触活动之前和之后将特定货物运输到脊柱中（例如通过 BDNF 诱导的可塑性或激活）在目标 3 中，我们将检验来自 FXS 患者的人类诱导多能干细胞 (hiPSC) 的神经元在 MT 和驱动蛋白侵入树突棘方面存在缺陷的假设。潜在的突触可塑性，可能对毁灭性的发育疾病产生影响。