Novel role of mitotic kinesin KIF11 in structural plasticity and memory

有丝分裂驱动蛋白 KIF11 在结构可塑性和记忆中的新作用

• 批准号: 10607093
• 负责人: Jenna Lynne Wingfield
• 金额: $ 6.76万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10607093
• 关键词: Alzheimer' s Disease; Architecture; Behavior; Behavioral Paradigm; Binding; Binding Proteins; Biological Assay; Brain; COX7A2L Protein; Cells; Data; Defect; Dendrites; Dendritic Spines; Development; Disease; Dorsal; Dynein ATPase; Electrophysiology (science); Ensure; Exhibits; Extinction (Psychology); Face; Frequencies; Genes; Glutamates; Growth; Health; Hippocampus (Brain); Human; Image; Intellectual functioning disability; Intracellular Transport; Kinesin; Learning; Learning Disorders; Lymphedema; Mediating; Mediator of activation protein; Memory; Mental Retardation; Microcephaly; Microtubule Proteins; Microtubules; Mitosis; Mitotic; Morphology; Motor; Mus; Mutation; Neurobiology; Neurons; Operative Surgical Procedures; Penetrance; Physiological; Protein Family; Proteins; RNA Interference; Regulation; Role; Synapses; Synaptic Transmission; Synaptic Vesicles; Synaptic plasticity; Synaptophysin; Syndrome; Testing; Training; Vertebral column; Work; autism spectrum disorder; density; experimental study; gain of function; in vivo; insight; knock-down; long term memory; loss of function; member; mental function; nervous system disorder; novel; overexpression; pleiotropism; polarized cell; preservation; protein function; synaptic function; therapeutic development; two photon microscopy

Project Summary Mitotic cells dynamically regulate microtubule (MT) architecture through the use of the MT motors: kinesin and dynein. Intriguingly, many of the canonical mitotic kinesins are expressed in post-mitotic neurons. These kinesins are thought to participate in repositioning MTs in neurons, yet their precise function and how they are regulated in neurons are unknown. Notably, mutations in kinesin-5 result in disorders associated with intellectual disabilities in humans, however, there is a lack of mechanistic understanding of why this occurs. Previously, the Puthanveettil lab revealed that members of the kinesin family of proteins (KIFs) are physiologically regulated in neurons and are required for long-term memory storage. Recent studies from the lab uncovered that KIF11 (Kinesin-5), acts as an inhibitory constraint on excitatory synaptic transmission in hippocampal neurons. RNAi- mediated loss-of-function analysis of KIF11 resulted in increased mini excitatory post-synaptic currents frequencies, increased dendritic branching, and increased synapse density. KIF11 is the only known homotetrameric kinesin, with four, slow motor domains that face outward to bind interpolar MTs during mitosis, and brake against other kinesins to ensure proper spindle formation. It is unknown how KIF11 mediates structural changes in hippocampal neurons, and whether KIF11 has a role in long-term memory storage. My central hypothesis is that KIF11 mediated regulation of MT dynamics is required for the activity-dependent structural changes in dendritic spines and long-term memory. To test this, in Aim1 I will uncover the mechanism behind how KIF11 regulates structural plasticity through examining activity-dependent changes in spine morphology following glutamate uncaging in WT, KIF11 KD, and KIF11 overexpression neurons by two-photon microscopy (2P) imaging while simultaneously visualizing MT dynamics via EB1. In Aim 2 I will test the effects of the loss of function and gain of function of KIF11 in dorsal CA1 neurons in learning and memory behavior assays. Completion of these aims will bring novel insights into the mechanism and regulation of KIF11 in structural plasticity and memory. Moreover, these studies will significantly advance our understanding of mitotic kinesins in post-mitotic neurons.

项目摘要 有丝分裂细胞通过使用MT电动机动态调节微管（MT）结构 动力蛋白。有趣的是，许多规范有丝分裂驱动蛋白在有丝分裂后神经元中表达。这些运动素 人们认为参与神经元中的MTS重新定位，但它们的精确功能以及如何调节它们 在神经元中是未知的。值得注意的是，运动蛋白5中的突变导致与智障相关的疾病 然而，在人类中，缺乏机械理解的理解。以前， Puthanveettil实验室透露，蛋白质的动力蛋白家族（KIF）在生理上受到调节 神经元，是长期记忆存储所必需的。实验室的最新研究发现了KIF11 （驱动蛋白5），对海马神经元中兴奋性突触传播的抑制作用。 rnai- 介导的KIF11功能丧失分析导致迷你兴奋性后突触后电流增加 频率，树突分支增加以及突触密度增加。 KIF11是唯一已知的 同型式驱动蛋白，有四个缓慢的运动结构域，它们在有丝分裂过程中向外朝外结合MT， 并针对其他驱动器制动，以确保正确的主轴形成。尚不清楚KIF11如何介导结构 海马神经元的变化以及KIF11是否在长期存储器中起作用。我的中央 假设是KIF11介导的MT动力学调节是活性依赖性结构所必需的 树突状刺和长期记忆的变化。为了测试这一点，在AIM1中，我将发现背后的机制 KIF11如何通过检查脊柱形态的活动依赖性变化来调节结构可塑性 在WT，KIF11 KD和KIF11的过表达神经元中，通过两光子显微镜检查后进行谷氨酸渗透。 （2p）成像，同时通过EB1同时可视化MT动力学。在AIM 2中，我将测试丢失的影响 在学习和记忆行为分析中，KIF11的功能和功能获得。 这些目标的完成将使KIF11在结构中的机制和调节中提供新的见解 可塑性和记忆。此外，这些研究将大大提高我们对有丝分裂驱动蛋白的理解 在有丝分裂后神经元中。