Role of 14-3-3epsilon in neurite initiation

项目总结/摘要 大脑中复杂神经元网络的形成需要精确的神经发生和神经元的发育。 在发育过程中迁移，然后形成神经突和棘。如果神经突和棘的形成 一旦被破坏，就会导致各种疾病，如精神疾病。最近，患有A 已经报道了17p13.3染色体区域的微复制，并且表现出17p13.3染色体区域的微复制的患者的数量是17p13.3染色体区域的17 p1 这种综合症一直在增加。有趣的是，患有17p13.3微重复综合征的患者 表现出严重的神经和形态缺陷，包括癫痫、智力迟钝和自闭症 谱系障碍（ASD）。更有趣的是，分析这种新的人类遗传综合征的研究澄清了 这一约70 kb的关键区域与ASD密切相关，并且该区域含有一个单一的 Ywhae基因编码14-3-3ε蛋白。这些数据有力地表明，Ywhae可能是负责 这些患者的ASD表型。14-3-3ε在神经元形态发生中的作用 形成，以前没有分析过，仍然未知。因此，这一目标 研究计划是分析神经突中14-3-3ε的新的体内功能和分子靶点 形成，特别是神经突的起始和延伸，并阐明这种细胞的分子机制， 活动我们假设14-3-3ε在神经突的起始和延伸中起着重要的作用， 通过与双皮质素（Dcx）的结合和微管亲和力控制微管（MT）滑动和稳定性 调节激酶3（Mark 3）。为了验证这一假设，我们建议使用各种各样的实验策略 包括子宫内电穿孔、体外和体内的延时实时成像、他莫昔芬诱导的Cre-loxP 系统和14-3-3ε条件性敲除小鼠。我们有三个具体目标。在具体目标1中，我们将测试 14-3-3ε调节神经突形成和突触发生的假说。在具体目标2中，假设 14-3-3ε通过与Dcx结合调节MT滑动来调节皮质中的神经突起始， 测试.在具体目标3中，我们将检验14-3-3ε通过结合 Mark 3并调节其活性。该项目的成功完成不仅将提供理解， 一种新的人类遗传综合征的病因学与ASD的过度表达有关， 14-3-3ε在神经突中的精确功能也显著增强了我们对14-3-3ε在体内功能的理解 启动和扩展。

Project Summary/Abstract The formation of the intricate neuronal network in the brain requires precise neurogenesis and neuronal migration followed by neurite and spine formation during development. If neurite and spine formation is disrupted, it results in a wide range of diseases such as mental illnesses. Recently, patients with a microduplication of the 17p13.3 chromosome region have been reported and the number of patients exhibiting this syndrome has been increasing. Interestingly, the patients with this 17p13.3 microduplication syndrome show severe neurological and morphological defects, including, epilepsy, mental retardation and autism spectrum disorders (ASD). More interestingly, the studies analyzing this new human genetic syndrome clarified that the critical region spanning about 70kb is strongly associated with ASD, and this region contains a single gene, Ywhae, encoding the protein 14-3-3ε. These data strongly suggest that Ywhae might be responsible for the ASD phenotype in these patients. The functions of 14-3-3ε in neuronal morphogenesis including neurite formation, have not been previously analyzed and still remain unknown. Therefore, the objectives of this research proposal are to analyze the novel in vivo functions and molecular targets of 14-3-3ε in neurite formation, in particular neurite initiation and extension, and clarifying the molecular mechanisms in this cellular event. We hypothesize that 14-3-3ε plays an important role in neurite initiation as well as neurite extension by controlling microtubule (MT) sliding and stability through binding to Doublecortin (Dcx) and Microtubule affinity regulating kinase 3 (Mark3). To test this hypothesis, we propose to use a wide variety of experimental strategies including in utero electroporation, time-lapse live imaging in vitro and in vivo, tamoxifen-inducible Cre-loxP system, and 14-3-3ε conditional knockout mice. We have three specific aims. In the Specific Aim 1, we will test the hypothesis that 14-3-3ε regulates neurite formation and synaptogenesis. In Specific Aim 2, the hypothesis that 14-3-3ε regulates neurite initiation in the cortex by regulating MT sliding through binding to Dcx will be tested. In Specific Aim 3, we will test the hypothesis that 14-3-3ε regulates neurite extension by binding to Mark3 and regulating its activity. The successful completion of this project will not only provide understanding as to the etiology of a new human genetic syndrome strongly associating with ASD caused by the overexpression of 14-3-3ε but also significantly enhance our understanding of the precise in vivo functions of 14-3-3ε in neurite initiation and extension.