Modulation of Microtubule Dynamics in Axon Guidance

轴突引导中微管动力学的调节

• 批准号: 8732123
• 负责人: Guofa Liu
• 金额: $ 44.25万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8732123
• 关键词: Address; Alzheimer' s Disease; Axon; Brain; Chemotactic Factors; Collaborations; Complex; Coupling; Cues; Cytoskeleton; Data; Defect; Development; Disease; Epilepsy; Event; Family; Growth Cones; Injury; Life; Link; Mediating; Microtubules; Nervous system structure; Neurons; Parkinson Disease; Play; Positioning Attribute; Post-Translational Protein Processing; Protein Isoforms; Proteins; Regulation; Role; Signal Pathway; Signal Transduction; Spinal Muscular Atrophy; Spinal cord injury; Stroke; Tubulin; Tyrosine Phosphorylation; axon growth; axon guidance; base; cellular imaging; design; human NTN1 protein; human disease; lissencephaly; netrin receptor; netrin-1; neuronal circuitry; periventricular heterotopia; public health relevance; receptor; research study; response; src-Family Kinases

DESCRIPTION (provided by applicant): Proper axon outgrowth and path finding are essential for the formation of functional neuronal circuits in the developing nervous system. Coordination of different guidance cues, their receptors and intracellular signal transduction cascades eventually converges on orchestrating cytoskeleton dynamics to maneuver growth cone navigation. Netrins, a conserved family of canonical guidance cues, play an important role in axon outgrowth and guidance in the developing nervous system. The coupling of axon guidance cues, such as netrin-1, to microtubule (MT) dynamics is crucial for growth cone (GC) navigation. However, whether axon guidance signaling regulates MT dynamics directly or indirectly is unclear. Our preliminary data indicate that DCC and DSCAM interact directly with TUBB3, the most dynamic ?-tubulin isoform in neurons, and netrin-1 induces these interactions. TUBB3 is required for netrin-1- induced axon outgrowth and path finding in the developing nervous system. These results suggest that DCC and DSCAM directly couple MT dynamics in netrin-1-mediated axon attraction. Interestingly, UNC5 also interacts with TUBB3. In this study, we will determine how netrin-1 modulates microtubule dynamics through coupling of its receptors DCC, DSCAM and UNC5C with dynamic TUBB3 in attractive and repulsive signaling. Aim1. Untangle the role of TUBB3 in coordination of netrin-1 attractive signaling. To further investigate the coordinating role of TUBB3 in netrin/DCC/DSCAM-mediated axon attractive signaling, we will examine: 1) Netrin-1 regulation of the interaction of TUBB3 with DCC and DSCAM; 2) Modulation of MT dynamics by netrin-1 via the interaction of dynamic TUBB3 with DCC and DSCAM; 3) The functional importance of TUBB3 in netrin attractive signaling. Aim 2. Determine the role of TUBB3 in coordination of netrin-1 repulsive signaling. UNC5 interacts with DCC and DSCAM and these interactions mediate repulsive responses to netrin-1. Our preliminary data show that TUBB3 interacts with UNC5C, DCC and DSCAM, suggesting it may play a role in coordinating netrin-1 repulsive signaling. To further characterize this hypothesis, we will focus on studying: 1) Netrin-1 regulation of the interaction of TUBB3 with UNC5C, DCC and DSCAM; 2) Modulation of MT dynamics by netrin-1 via coordination of the interaction of dynamic TUBB3 with UNC5C, DCC and DSCAM; 3) The functional importance of TUBB3 in netrin-1-mediated axon repulsion. Aim 3. Investigate the mechanistic link between netrin signaling and MT dynamics. To examine how netrin-1 directly regulates MT dynamics via TUBB3 in axon turning, we will focus on: 1) assessing tyrosine phosphorylation of TUBB3 in netrin signaling; 2) tracking modulation of MT dynamics in the GC during netrin-1-mediated attraction using live cell imaging; 3) tracking netrin-1- mediated MT dynamics in the GC during axon repulsion. This study should reveal how netrin-1 directly modulates MT dynamics via differential coupling of its receptors with dynamic TUBB3 in axon turning.

描述（由申请人提供）：在发育中的神经系统中，适当的轴突生长和寻径对于功能性神经元回路的形成至关重要。不同的引导信号，它们的受体和细胞内信号转导级联的协调最终汇聚在协调细胞骨架动力学来操纵生长锥导航。神经网络是一类保守的典型引导信号，在神经系统发育的轴突生长和引导中起着重要作用。轴突引导信号（如netrin-1）与微管（MT）动力学的耦合对于生长锥（GC）导航至关重要。然而，轴突引导信号是否直接或间接调节MT动力学尚不清楚。我们的初步数据表明，DCC和DSCAM直接与最具活力的TUBB3相互作用。-微管蛋白异构体和netrin-1诱导了这些相互作用。在发育中的神经系统中，TUBB3是由netrin-1诱导的轴突生长和寻径所必需的。这些结果表明DCC和DSCAM在netrin-1介导的轴突吸引中直接耦合MT动力学。有趣的是，UNC5也与TUBB3相互作用。在本研究中，我们将确定netrin-1如何通过其受体DCC、DSCAM和UNC5C与动态TUBB3在吸引和排斥信号传导中的偶联来调节微管动力学。Aim1。解开TUBB3在协调netrin-1吸引信号中的作用。为了进一步研究TUBB3在netrin/DCC/DSCAM介导的轴突吸引信号传导中的协调作用，我们将研究：1)netrin -1调节TUBB3与DCC和DSCAM的相互作用；2) netrin-1通过动态TUBB3与DCC和DSCAM的相互作用调制MT动力学；3) TUBB3在网络蛋白吸引信号传导中的功能重要性。目标2。确定TUBB3在协调netrin-1排斥性信号传导中的作用。UNC5与DCC和DSCAM相互作用，这些相互作用介导对netrin-1的排斥反应。我们的初步数据显示，TUBB3与UNC5C、DCC和DSCAM相互作用，表明它可能在协调netrin-1排斥信号传导中发挥作用。为了进一步表征这一假设，我们将重点研究：1)Netrin-1调控TUBB3与UNC5C、DCC和DSCAM的相互作用；2)通过协调动态TUBB3与UNC5C、DCC和DSCAM的相互作用，netrin-1对MT动力学的调制；3) TUBB3在netrin-1介导的轴突排斥中的功能重要性。目标3。研究神经网络信号传导和MT动力学之间的机制联系。为了研究netrin-1如何通过TUBB3在轴突转向中直接调节MT动力学，我们将重点关注：1)评估netrin信号中TUBB3的酪氨酸磷酸化；2)利用活细胞成像跟踪调制netrin-1介导的吸引过程中GC中MT动力学；3)在轴突排斥过程中跟踪神经网络-1介导的GC MT动力学。这项研究将揭示netrin-1如何通过其受体与动态TUBB3在轴突转动中的差异偶联直接调节MT动力学。

项目成果

Human TUBB3 Mutations Disrupt Netrin Attractive Signaling.

• DOI: 10.1016/j.neuroscience.2018.01.046
• 发表时间: 2018-03-15
• 期刊: Neuroscience
• 影响因子: 3.3
• 作者: Huang H;Yang T;Shao Q;Majumder T;Mell K;Liu G
• 通讯作者: Liu G

Coordinated interaction of Down syndrome cell adhesion molecule and deleted in colorectal cancer with dynamic TUBB3 mediates Netrin-1-induced axon branching.

• DOI: 10.1016/j.neuroscience.2015.02.042
• 发表时间: 2015-05-07
• 期刊: Neuroscience
• 影响因子: 3.3
• 作者: Huang H;Shao Q;Qu C;Yang T;Dwyer T;Liu G
• 通讯作者: Liu G

Disease-associated mutations in human TUBB3 disturb netrin repulsive signaling.

人类 TUBB3 中与疾病相关的突变会扰乱 netrin 排斥信号。

• DOI: 10.1371/journal.pone.0218811
• 发表时间: 2019
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Shao,Qiangqiang;Yang,Tao;Huang,Huai;Majumder,Tanushree;Khot,BhaktiAjit;Khouzani,MohammadMasoudian;Alarmanazi,Farrah;Gore,YasminK;Liu,Guofa
• 通讯作者: Liu,Guofa