Development and afferent regulation of auditory neurons

项目摘要 神经元树突的形态和固有特性是其功能的专门化。树枝状缺陷是 与许多神经发育障碍密切相关。在这个提案中，我努力确定角色 脆性X智力低下蛋白(FMRP)在听神经元树突调节中的作用。缺少FMRP 结果导致脆性X综合征(FXS)，这是自闭症最常见的遗传性单基因原因，表现为 包括智力缺陷和感觉障碍在内的一系列症状。我建议研究FMRP 脑干双耳回路发育过程中的调控及其在脑发育中的作用 传入活性和完整性改变后树突状细胞形态和生化的调节。 我将对鸡的大细胞核(NM)和板层核(NL)进行详细的分析。 用于研究听觉时间加工的特征良好的动物模型和易于处理的基因系统 操作，并将一些分析扩展到人脑。 我将用免疫印迹和免疫印迹技术研究鸡NM和NL中FMRP的发育特征。 免疫细胞化学，并确定FMRP与良好记录的时间相关性 树突发育的特点和特殊的生理特性。 我将确定FMRP表达下调如何影响树突状细胞形态的发展 以及鸡NM和NL神经元关键蛋白的表达。利用时间和时间进行基因操作 空间控制、单个细胞填充、免疫细胞化学和高分辨率共聚焦显微镜将 被利用。 我将确定FMRP如何调控鸡NL中依赖传入的树突状细胞重组。这就做 首先研究传入如何调节FMRP，然后评估FMRP基因敲除对 树突状结构和生化的传入影响。结合了遗传和传入的操作， 以及固定和活组织的共焦和多光子成像。 ？与Kulesza博士合作，我们将开始探索FMRP在人类听觉中的潜在功能 检测FMRP及其结合的脑干神经元的时空分布 用免疫细胞化学方法在固定的人脑干切片中检测蛋白质。 总体而言，这些研究将为FMRP对树突分枝和特化的调控提供洞察力 听觉处理神经元的特性和脊椎动物神经元发育机制的洞察 和疾病病理学。

Project Summary Neuronal dendritic morphology and intrinsic properties are specialized for their function. Dendritic defects are strongly associated with numerous neurodevelopmental disorders. In this proposal, I endeavor to identify roles of fragile X mental retardation protein (FMRP) in dendritic regulation of auditory neurons. Absence of FMRP results in fragile X syndrome (FXS), the most frequent inherited monogenetic cause of autism, presenting with a constellation of symptoms that include intelligence deficits and sensory dysfunction. I propose to study FMRP regulation during development of very well characterized binaural circuitry in the brainstem and its role in regulation of dendritic morphology and biochemistry following changes in afferent activity and integrity. I will conduct detailed analyses in the chicken nucleus magnocellularis (NM) and nucleus laminaris (NL), a well-characterized animal model for studying auditory temporal processing and a tractable system for gene manipulation, and will extend some analyses to human brains. ¿ I will characterize the developmental profile of FMRP in chicken NM and NL using Western blot and immunocytochemistry, and identify the temporal correlations of FMRP with well-documented characteristics of dendritic development and specialized physiological properties. ¿ I will determine how knockdown of FMRP expression affects the development of dendritic morphology and expression of key proteins of chicken NM and NL neurons. Gene manipulations with temporal and spatial control, individual cell filling, immunocytochemistry, and high-resolution confocal microscopy will be used. ¿ I will determine how FMRP regulates afferent-dependent dendritic reorganization in chicken NL. I will first examine how afferent regulates FMRP and then assess the effects of FMRP knockdown on afferent influence of dendritic structure and biochemistry. Combined genetic and afferent manipulations, as well as confocal and multi-photon imaging of fixed and live tissues, will be used. ¿ In collaboration with Dr. Kulesza, we will start to explore potential function of FMRP in human auditory brainstem neurons by examining the temporal and spatial distribution of FMRP and FMRP-binding proteins in fixed human brainstem sections using immunocytochemistry. Overall, these studies will provide insight into FMRP regulation of dendritic arborization and specialized properties for auditory processing neurons, and insight into mechanisms of vertebrate neuronal development and disease pathology.