NEURITE OUTGROWTH AND THE ACQUISITION OF NEURITE IDENTIT

该提案旨在实现拉丁美洲倡议的目标 来自福格蒂国际研究合作计划，通过寻求 支持继续在美国开始的合作 目前正在阿根廷继续进行。 协作工作形成了 在此提议的基础上开发了反义新技术 寡核苷酸抑制微管相关蛋白的表达 蛋白质 (MAP)、tau。 随后该工作被广泛引用 做出重要的技术贡献并展示细胞 神经元 MAP 在神经突生长中的生物学功能。 从那时起， 合作工作仍在继续，其中使用反义技术 已扩展到解决运动蛋白的重要领域。 的 神经元必须沿着微管输送分子的长距离 轨道使这些蛋白质成为神经突生长的重要方面 神经发育和再生。 培养中的小脑大神经元和海马锥体细胞将 用于研究神经突生长的分子基础。 专注于 MAP，在神经元的顺序发育中发挥作用 形态特征，我们将使用反义寡核苷酸来创建 有丝分裂后神经元的无效表型。 反义的特异性 寡核苷酸允许选择性抑制单个 mRNA 及其 翻译产品。 我们计划分析其他 MAP，特别是 MAP1B 和基于微管的运动蛋白驱动蛋白。 通过结合 对反义处理的无效细胞进行直接视频显微镜分析 形态测量、免疫细胞化学和超微结构图像，我们希望 在神经突发生过程中为这些蛋白质分配离散的功能。

This proposal is intended to meet the aims of the Latin American Initiative from the Fogarty International Research Collaboration program by seeking support for the continuation of a collaboration begun in the U.S. and currently continuing in Argentina. The collaborative work which forms the basis of this proposal developed the novel technique of antisense oligonucleotides to suppress the expression of the microtubule-associated protein (MAP), tau. Subsequently this work has been widely cited for making an important technical contribution and for demonstrating a cell biological function of a neuronal MAP in neurite outgrowth. Since then, collaborative work has continued in which the use of antisense techniques have been expanded to address the important area of motor proteins. The long distances over which neurons must deliver molecules along microtubule tracks makes these proteins an essential aspect of neurite growth in both neural development and in regeneration. Cerebellar macroneurons and hippocampal pyramidal cells in culture will be used to study the molecular basis of neurite outgrowth. Focusing upon the MAPs, which have a role in the sequential development of neuronal morphological features, we will use antisense oligonucleotides to create null phenotypes in post-mitotic neurons. The specificity of the antisense oligonucleotides permits the selective suppression of a single mRNA and its translation product. We plan to analyze additional MAPs, particularly MAP1B and the microtubule-based motor protein, kinesin. By combining direct video-microscopic analyses of antisense-treated null cells with morphometric, immunocytochemical, and ultrastructural images, we hope to assign discrete functions to these proteins during neuritogenesis.