SPECTRIN-LIKE PROTEIN IN DEVELOPING BRAIN

该项目旨在探索幽灵蛋白和相关的 细胞骨架蛋白与细胞结构和功能的关系 发育中的神经系统。在这件事的前两年 格兰特我们发现了两种脑影蛋白亚型(240/235) 和(240/235E)，并对它们进行了定位 免疫细胞化学和免疫电子技术的异构体 显微镜。脑影蛋白(240/235)主要位于 神经元的轴突和突触前终末。脑部光谱 (240/235E)存在于胞体、树突和突触后 神经元的终末，以及某些类型的神经胶质细胞。这个 这些血影蛋白亚型的个体发育在 免疫放射自显影实验，并与 免疫细胞化学程序。每一种血影蛋白亚型都有一个 在发展中表现和分布的独特模式 神经系统。在本申请中，我们描述了实验 旨在让我们更好地了解大脑 血影蛋白亚型及相关的血影蛋白结合蛋白 关于哺乳动物神经系统的发育。 这项建议的目的是：(1)确定选址和 发育过程中小鼠脑内血影蛋白亚型的分布 用免疫电子显微镜观察脑组织。(2)量化大脑 血影蛋白(240/235E)用免疫斑点定量分析。(3) 定义在同种异构体中是否存在结构变化 在发育过程中使用免疫沉淀和 多肽图谱技术。(4)检查是否有两个 哺乳动物脑内Ankyrin(Syndein)亚型的利用 免疫印迹和免疫组织化学。(5)确定 用免疫印迹法研究脑内锚蛋白亚型的个体发育 免疫斑点、免疫组织化学和免疫电子 显微镜。(6)检测Amelin在大鼠脑内的表达。 使用概述的技术的哺乳动物大脑发育 在(5)中。所提出的研究构成了重要而新颖的内容 血影蛋白样蛋白功能意义的探讨 在神经细胞和组织的结合蛋白中。这 调查是正在进行的蜂窝和 分子神经生物学，试图了解 正常和异常的大脑发育。

This project is designed to explore spectrin and associated cytoskeletal proteins in regard to the structure and function of the developing nervous system. During the first 2 years of this grant we have discovered two subtypes of brain spectrin (240/235) and (240/235E) in the mammalian brain, and localized these isoforms with immunocytochemistry and immunoelectron microscopy. Brain spectrin (240/235) is located primarily in the axons and presynaptic terminals of neurons. Brain spectrin (240/235E) is found in the cell bodies, dendrites, and postsynaptic terminals of neurons, as well as in certain glial cell types. The ontogeny of these spectrin subtypes was explored in immunoautoradiography experiments, and closely examined with immunocytochemical procedures. Each spectrin subtype had a distinct pattern of expression and distribution in the developing nervous system. In this application we describe experiments which are designed to give us a better understanding of brain spectrin subtypes and associated spectrin binding proteins in regard to the development of the mammalian nervous system. The aims of this proposal are: (1) Determine the location and distribution of brain spectrin subtypes in the developing mouse brain using immunoelectron microscopy. (2) Quantitate brain spectrin (240/235E) with quantitative immunodot assay. (3) Define whether there are structural changes in the isoforms of spectrin during development using immunoprecipitation and peptide mapping techniques. (4) Examine whether there are two ankyrin (syndein) subtypes in the mammalian brain utilizing immunoblots and immunohistochemistry. (5) Determine the ontogeny of ankyrin subtypes in the brain with immunoblots, immunodots, immunohistochemistry, and immunoelectron microscopy. (6) Examine the expression of amelin during mammalian brain development employing the techniques outlined in (5). The studies proposed constitute important and novel inquiries into the functional significance of spectrin-like proteins in the binding proteins in neural cells and tissues. This investigation is part of an ongoing program in cellular and molecular neurobiology which seeks to understand the process of normal and abnormal brain development.