DEVELOPMENT AND MIGRATION OF LHRH NEURONS

我目前的职业目标是继续过渡到 分子发育神经科学。我的长期目标是成为 像我一样熟悉解决发育问题的分子方法 我学的是神经解剖学和胚胎学。这两个领域的专业知识将是 在评估基因在发育过程中的功能方面很有用。目前， 教学和行政工作占据了我相当大的一部分 时间到了。如果L获得了RCDA，将被免除许多这些职责。 这将腾出时间，让我可以收到额外的 分子技术方面的培训。托马斯·罗森奎斯特博士，我的 部门主席非常支持我的RCDA申请。 如果我收到这笔赠款，他已经表示他将提供资金 对于一个有分子生物学经验的博士后来说 在我的实验室里。细胞生物学和解剖学系有 最近成立了一个分子生物学核心设施。这个设施将 包含许多我需要的资源，以完成建议的 实验。此外，L还将利用单抗 内布拉斯加州大学医学中心的设施。在这个RCDA中 建议，我们计划利用我们对LHRH的熟悉 大鼠嗅觉系统发育与神经元迁移 鸡来研究LHRH神经元在发育过程中是如何进入大脑的。 最近的研究表明，X连锁Kallmann病是 由于嗅神经未能进入端脑和 LHRH神经元迁移到大脑中。与此相关的基因 疾病，KAL，可能是第一个被发现有作用的人类基因 在神经元迁移中。我们有三个具体的目标：瞄准L。 嗅神经在LHRH神经元迁移中的作用在这个特定的目标中 我们试图回答两个问题。首先，LHRH神经元是否会沿着 嗅神经的短暂延伸？第二，LHRH神经元 与嗅神经中相同的轴突相联系 移民？我们希望通过给嗅觉贴上标签来回答这些问题 处于不同发育阶段的神经轴突。Aim Il.空间- KAL蛋白在光和电子状态下的时间分布 微观层面。因为似乎存在时空不匹配 在KAL消息和LHRH神经元迁移之间，关键是 检测KAL蛋白在发育过程中的分布。我们将使用 双标记免疫细胞化学检测LHRH神经元 迁移与KAL蛋白表达相关。目标三.职能 KAL蛋白的。由于KAL的突变扰乱了LHRH神经元 迁移和嗅神经寻路，有很好的理由 相信KAL参与了这些事件。然而，这种机制 因为这些影响是未知的。我们将使用组织培养，消融， 并通过抗体和多肽阻断实验来确定其作用 卡尔在神经系统发育中的作用。独一无二的机遇 在Chick系统中可用，并且事实是完整的编码 KAL的mRNA序列在雏鸡中是可用的，使这成为 最佳Kallmann综合征动物模型系统。此外，这些 研究将使我们能够剖析神经元迁移和 可能适用于大脑其他区域的轴突路径发现 发育中的神经系统。

My immediate career objective is to continue the transition to molecular developmental neuroscience. My long term goal is to become as familiar with molecular approaches to developmental problems as I am with neuroanatomy and embryology. Expertise in both areas will be useful in evaluating the function of genes in development. Currently, teaching and administrative duties take up a considerable amount of my time. If l received a RCDA, would be relieved of many of these duties. This would free up time that would allow me to receive additional training in molecular techniques. Dr. Thomas Rosenquist, my Departmental Chairman, is very supportive of my application for a RCDA. If I receive this grant, he has indicated that he will provide funds for a post-doctoral fellow with experience in molecular biology to work in my laboratory. The department of Cell Biology and Anatomy has recently formed a Molecular Biology Core Facility. This facility will contain many of the resources I need to complete the proposed experiments. In addition, l will also utilize the Monoclonal Antibody Facility at the University of Nebraska Medical Center. In this RCDA proposal, we plan to take advantage of our familiarity with LHRH neuronal migration and the development of the olfactory system in the chick to examine how LHRH neurons enter the brain during development. Recent studies have demonstrated that X-linked Kallmann's disease is due to a failure of the olfactory nerve to enter the telencephalon and LHRH neurons to migrate into the brain. The gene involved in this disease, KAL, may be the first human gene identified as having a role in neuronal migration. We have three specific aims: Aim l. The role of the olfactory nerve in LHRH neuronal migration. In this specific aim we seek to answer two questions. First, do LHRH neurons migrate along a transitory extension of the olfactory nerve? Second, are LHRH neurons associated with the same axons in the olfactory nerve throughout their migration? We hope to answer these questions by labeling olfactory nerve axons at different stages of development. Aim Il. Spatial- temporal distribution of the KAL protein at the light and electron microscopic levels. As there appears to be a spatial-temporal mismatch between KAL message and LHRH neuronal migration, it is crucial to examine the distribution of KAL protein during development. We will use double-label immunocytochemistry to determine whether LHRH neuronal migration is correlated with KAL protein expression. Aim III. Function of the KAL protein. Since mutations in KAL disrupt LHRH neuronal migration and olfactory nerve pathfinding, there is good reason to believe that KAL is involved in these events. However, the mechanism for these effects is not known. We will use tissue culture, ablation, and antibody and peptide blocking experiments to determine the role of KAL in the development of the nervous system. The unique opportunities available in the chick system, and the fact that the complete coding sequence of the mRNA for KAL is available in the chick, makes this the best animal model system for Kallmann's syndrome. In addition, these studies will allow us to dissect mechanisms for neuronal migration and axon pathfinding that may be applicable to other regions of the developing nervous system.