Localized translation of cytoskeletal proteins in axonal morphogenesis

细胞骨架蛋白在轴突形态发生中的局部翻译

• 批准号: 8577831
• 负责人: GIANLUCA GALLO
• 金额: $ 33.91万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8577831
• 关键词: Actins; Address; Adult; Affect; Animal Model; Axon; Biological Models; Cell Adhesion Molecules; Cells; Cellular biology; Chick Embryo; Code; Complex; Cytoskeletal Proteins; Cytoskeleton; Data; Development; Disease; Embryo; Extracellular Matrix; Filopodia; Fluorescent in Situ Hybridization; Foundations; Future; Genetic Translation; Goals; Growth Cones; Image; Image Analysis; In Vitro; Individual; Investigation; Joints; Lead; Life; Link; Mediating; Mediator of activation protein; Messenger RNA; Methods; Microfluidics; Morphogenesis; Nerve Growth Factors; Nervous System Trauma; Nervous system structure; Neuronal Injury; Neuronal Plasticity; Neurons; Organism; Pattern; Phosphatidylinositols; Phosphotransferases; Preparation; Protein Biosynthesis; Proteins; Reagent; Recovery of Function; Regulation; Reporter; Reverse Transcriptase Polymerase Chain Reaction; Role; Semaphorin-3A; Sensory; Set protein; Signal Transduction; Small Interfering RNA; Spinal Cord; Synapses; System; Testing; Therapeutic; Time; Translating; Translations; Work; axon growth; base; experimental analysis; extracellular; frontier; human EMS1 protein; in vivo; microtubule-associated protein 1B; neuronal cell body; novel; novel strategies; public health relevance; tool

DESCRIPTION (provided by applicant): The formation of axon collateral branches underlies the ability of neurons to make synaptic contacts with multiple target neurons and thus give rise to complex neuronal networks. In the context of nervous system injury, the formation of axon collateral branches can have either beneficial or undesired effects depending on the affected circuitry. Collateral branches are also affected by a variety of disease states. However, the cellular mechanisms of collateral branching are only minimally understood. The preliminary data presented in this proposal unveils for the first time that the branch-inducing signal nerve growth factor drives the intra-axonal synthesis of cytoskeletal proteins with roles in the formation of collateral branches. Importantly, the preliminary data also demonstrate that axonal protein synthesis is required for the induction of axon collateral branching by nerve growth factor. Although recent studies have revealed a large set of mRNAs targeted to axons, the functional roles of the axonal translation of axonal mRNAs remains minimally understood. The main aim of the proposal is to determine the roles of the axonal synthesis of individual cytoskeletal proteins in the formation of axon collateral branches. By bridging the expertise of the PI (Dr. Gallo; neuronal cytoskeletal cell biology) and the Co-I (Dr. Twiss; axonal protein synthesis), the project provides the unique opportunity to uncover a new aspect of the mechanism of axon branching and aims to directly link the axonal synthesis of specific cytoskeletal proteins to the regulation f the dynamics of the axonal cytoskeleton. Through the joint expertise of the PI and Co-I, the project takes a multi-pronged in vivo and in vitro approach to address the main Aims. Collateral branching is affected by nervous system injury and disease. However, the ability to manipulate branching in a therapeutic context is mostly lacking. The project has the potential to lead to strategies for the regulation of axon branching by targeting specific axonal mRNA species in the context of neuronal injury and disease. The reagents developed for use in the project (e.g., cell permeable tools to selectively inhibit the axonal translation of individual mRNA species) have the potential for translation to animal model system of collateral branching in future directions o the project. The work we propose will serve as the foundation for these long term goals by determining the relevant mRNA targets through the elucidation of their roles in branching.

描述（由申请人提供）：轴突侧支的形成是神经元与多个靶神经元进行突触接触的能力的基础，从而产生复杂的神经元网络。在神经系统损伤的情况下，轴突侧支的形成可以具有有益或不期望的效果，这取决于受影响的回路。侧枝也受到各种疾病状态的影响。然而，侧支的细胞机制只有最低限度的了解。该提案中提出的初步数据首次揭示了分支诱导信号神经生长因子驱动轴突内细胞骨架蛋白的合成，并在侧支形成中发挥作用。重要的是，初步的数据还表明，轴突蛋白质合成所需的诱导轴突侧支神经生长因子。虽然最近的研究已经揭示了大量的靶向轴突的mRNA，轴突mRNA的轴突翻译的功能作用仍然知之甚少。该提案的主要目的是确定轴突合成的作用，个别细胞骨架蛋白的轴突侧支的形成。通过桥接PI（Gallo博士;神经元细胞骨架细胞生物学）和Co-I（Twiss博士;轴突蛋白合成）的专业知识，该项目 为揭示轴突分支机制的新方面提供了独特的机会，并旨在将特定细胞骨架蛋白的轴突合成与轴突细胞骨架动力学的调节直接联系起来。通过PI和Co-I的联合专业知识，该项目采用多管齐下的体内和体外方法来实现主要目标。侧支受神经系统损伤和疾病的影响。然而，在治疗背景下操纵分支的能力大多缺乏。该项目有可能导致的策略，通过针对特定的轴突mRNA物种的神经元损伤和疾病的情况下，轴突分支的调节。为项目中使用而开发的试剂（例如，选择性抑制单个mRNA种类的轴突翻译的细胞可渗透工具）在该项目的未来方向中具有翻译到侧枝分支的动物模型系统的潜力。我们提出的工作将作为这些长期目标的基础，通过阐明它们在分支中的作用来确定相关的mRNA靶点。