Role of PAK5 in neurons and dendritic spines

PAK5 在神经元和树突棘中的作用

• 批准号: 7371038
• 负责人: AUDREY G MINDEN
• 金额: $ 29.56万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7371038
• 关键词: Actins; Biolistics; Biological Sciences; Brain; Cell Line; Cells; Cellular Morphology; Collaborations; Cultured Cells; Cytoskeletal Modeling; Cytoskeleton; Dendrites; Dendritic Spines; Development; Family; Fibroblasts; Filopodia; Goals; Investigation; Knockout Mice; Life; Mediating; Microscopy; Molecular; Molecular Target; Morphology; Mus; Neocortex; Neurites; Neuroblastoma; Neurons; PAK6 gene; Pathway interactions; Play; Protein-Serine-Threonine Kinases; Proteins; Purkinje Cells; Regulation; Role; Shapes; Slice; Structure; Surface; Synapses; System; Techniques; Technology; Thinking; Tissues; Transfection; Universities; Vertebral column; Work; axon guidance; cell motility; density; experience; genetic regulatory protein; hippocampal pyramidal neuron; member; novel; research study; rho; rho GTP-Binding Proteins; two-photon

DESCRIPTION (provided by applicant): The Rho GTPases Cdc42, Rac, and Rho regulate cell morphology by controlling the organization of the actin cytoskeleton. While they were originally characterized in fibroblasts, they also have key roles in neuronal development. For example, they play important roles in controlling axon guidance and neurite outgrowth. Our recent work has indicated that another function of the Rho GTPases may be to control the densities and morphologies of dendritic spines. Dendritic spines are actin rich projections found on the surface of most dendrites, and they mediate most synaptic contacts in the brain. The regulation of spine density, shape, and motility is thought to have important implications for the development and function of the neocortex. The targets for the Rho GTPases that mediate changes in neuronal cell morphology are not entirely known. We have identified a new target for Cdc42 and Rac called PAK5, which is a serine/threonine kinase that is expressed primarily in the brain. We have found that PAK5 triggers filopodia formation and neurite outgrowth in neuronal cell lines. The substrates for PAK5 are unknown. The goal of this proposal is to identify PAK5 targets and regulatory proteins and to determine whether and how PAK5 and its target proteins control neuronal morphology. In the first aim we will identify molecular targets for PAK5 and proteins that interact with PAK5. In the second aim we will determine whether these proteins are part of the pathway that leads to neurite outgrowth and filopodia formation in N1E-115 cells. In the third aim we will use biolistic transfections and two photon microscopy to investigate the possible role for PAK5 in regulating the morphology, density, and motility of dendritic spines in pyramidal neurons and purkinje cells from mouse brain slices. In this aim we will also examine spine motility and structure in PAK5 knockout mice that are being generated in our lab. Overall, the results from the experiments in this proposal will help elucidate the molecular mechanisms that control neuronal cell morphology and spinogenesis, which have important repercussion for the development and function of the normal and diseased brain.

描述(由申请人提供)： Rho GTP酶CDC42、Rac和Rho通过控制肌动蛋白细胞骨架的组织来调节细胞形态。虽然它们最初以成纤维细胞为特征，但它们在神经元发育中也扮演着关键角色。例如，它们在控制轴突引导和轴突生长方面发挥着重要作用。我们最近的工作表明，Rho GTP酶的另一个功能可能是控制树突棘的密度和形态。树突棘是在大多数树突表面发现的富含肌动蛋白的突起，它们介导了大脑中大多数突触的联系。脊椎密度、形状和运动性的调节被认为对新皮质的发育和功能具有重要意义。调节神经细胞形态变化的Rho GTP酶的靶标尚不完全清楚。我们已经为CDC42和RAC确定了一个新的靶点，称为PAK5，它是一种丝氨酸/苏氨酸激酶，主要在大脑中表达。我们已经发现PAK5在神经细胞系中触发丝状足突的形成和轴突的生长。PAK5的底物尚不清楚。该提案的目的是确定PAK5的靶点和调控蛋白，并确定PAK5及其靶蛋白是否以及如何控制神经元的形态。在第一个目标中，我们将确定PAK5的分子靶点以及与PAK5相互作用的蛋白质。在第二个目标中，我们将确定这些蛋白质是否是导致N1E-115细胞轴突生长和丝状足形成的途径的一部分。在第三个目标中，我们将使用生物转基因和双光子显微镜来研究PAK5在调节来自小鼠脑片的锥体神经元和浦肯野细胞中树突棘的形态、密度和运动方面的可能作用。在这个目标中，我们还将检查我们实验室培育的PAK5基因敲除小鼠的脊柱运动和结构。总体而言，这项提议中的实验结果将有助于阐明控制神经细胞形态和棘突发生的分子机制，这些机制对正常和疾病大脑的发育和功能具有重要影响。