SPECTRIN-LIKE PROTEIN IN DEVELOPING BRAIN

该项目旨在探索血影蛋白和相关的 细胞骨架蛋白的结构和功能 发育中的神经系统 在最初的两年里， 我们发现了两种脑血影蛋白亚型（240/235） 和（240/235 E）在哺乳动物脑中的分布，并定位了这些 免疫细胞化学和免疫电子学 显微镜 脑血影蛋白（240/235）主要位于 神经元的轴突和突触前末梢。 脑血影蛋白 (240/235 E）存在于细胞体、树突和突触后 神经元的末端，以及某些神经胶质细胞类型。 的 这些血影蛋白亚型的个体发生进行了探讨， 免疫放射自显影实验，并仔细检查 免疫细胞化学方法。 每种血影蛋白亚型都有一个 不同的表达和分布模式， 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 血影蛋白亚型和相关血影蛋白结合蛋白 关于哺乳动物神经系统的发展。 本建议的目的是：（1）确定地点， 发育期小鼠脑血影蛋白亚型的分布 大脑的免疫电子显微镜。 (2)定量脑 血影蛋白（240/235 E）定量免疫斑点法。 （三） 定义是否存在异构体的结构变化 血影蛋白在显影过程中使用免疫沉淀， 肽图谱技术。 (4)检查是否有两个 锚蛋白（syndein）亚型在哺乳动物大脑中利用 免疫印迹和免疫组织化学。 (5)确定 用免疫印迹分析脑中锚蛋白亚型的个体发生， 免疫点、免疫组织化学和免疫电子 显微镜 (6)检查amelin的表达 哺乳动物的大脑发育采用的技术概述 在（5）中。 提出的研究构成了重要和新颖的 血影蛋白样蛋白的功能意义探讨 神经细胞和组织中的结合蛋白。 这 调查是正在进行的蜂窝计划的一部分， 分子神经生物学试图理解 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.