DEVELOPMENT AND MIGRATION OF LHRH NEURONS

我职业生涯的近期目标是继续向 分子发育神经科学我的长期目标是成为 和我一样熟悉研究发育问题的分子方法 神经解剖学和胚胎学。这两个领域的专业知识将 用于评估基因在发育中的功能。目前， 教学和行政工作占用了我相当大的时间。 时间如果我得到一个RCDA，将被免除许多这些职责。 这将腾出时间，使我能够收到更多的 分子技术培训。托马斯罗森奎斯特博士，我的 部门主席非常支持我申请RCDA。 如果我收到这笔赠款，他已表示，他将提供资金 一个有分子生物学经验的博士后研究员 在我的实验室里细胞生物学和解剖学系 最近成立了一个分子生物学核心机构该设施将 包含许多我需要的资源，以完成拟议的 实验另外，我还将利用单克隆抗体 内布拉斯加大学医学中心的设施。在这个RCDA 我们计划利用我们对LHRH的熟悉， 神经元迁移和嗅觉系统的发育 研究LHRH神经元如何在发育过程中进入大脑。 最近的研究表明，X连锁卡尔曼氏病是 由于嗅觉神经未能进入端脑， LHRH神经元迁移到大脑。与此相关的基因 疾病，KAL，可能是第一个被确定为具有作用的人类基因。 在神经元迁移中。我们有三个具体目标：目标1。的作用 嗅神经在LHRH神经元迁移中的作用在这一具体目标中， 我们想回答两个问题。首先，LHRH神经元是否沿着沿着 嗅觉神经的短暂延伸其次，LHRH神经元 与嗅觉神经中的相同轴突相关， 迁移吗？我们希望通过标记嗅觉来回答这些问题 不同发育阶段的神经轴突。艾姆岛空间- KAL蛋白在光和电子下的时间分布 微观层面。因为这似乎是一个时空不匹配 在KAL信息和LHRH神经元迁移之间， 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，以确定是否LHRH神经元 迁移与KAL蛋白表达相关。Aim III.功能 KAL蛋白质。由于KAL突变破坏了LHRH神经元 迁移和嗅觉神经寻路，有很好的理由 相信KAL与这些事件有关然而，机制 这些影响是未知的。我们会用组织培养，消融， 以及抗体和肽阻断实验，以确定 KAL在神经系统的发展。的独特机会 可以在小鸡系统中使用，而且事实上， KAL的mRNA序列在小鸡中是可用的，这使得它成为 Kallmann综合征的最佳动物模型系统。另外这些 研究将使我们能够剖析神经元迁移的机制， 轴突寻路，这可能适用于其他地区的 神经系统发育。

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.