NEURITE OUTGROWTH AND THE ACQUISITION OF NEURITE IDENTIT

这项提议旨在实现拉丁美洲倡议的目标 从Fogarty国际研究合作计划中寻求 支持继续在美国和 目前仍在阿根廷继续。协作工作构成了 在这一提议的基础上，发展了反义新技术 寡核苷酸抑制微管相关基因表达的研究 蛋白质(MAP)、tau。随后，这项工作被广泛引用为 做出了重要的技术贡献，并展示了一个电池 神经元MAP在轴突生长中的生物学功能。从那以后， 合作工作仍在继续，其中反义技术的使用 已经扩展到解决发动机蛋白这一重要领域。这个 神经元必须沿着微管运送分子的长距离 Traces使这些蛋白质成为两者中神经突起生长的一个重要方面。 神经发育和再生。 小脑大神经元和海马区锥体细胞将在培养中 用于研究轴突生长的分子基础。专注于 MAP，它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零表型。反义基因的专一性 寡核苷酸允许选择性地抑制单个mRNA及其 翻译产品。我们计划分析更多的地图，特别是 MAP1B和基于微管的运动蛋白Kinesin。通过组合 反义处理的空细胞的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 在神经发生过程中为这些蛋白质分配不同的功能。

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.