MOLECULAR GENETIC ANALYSIS OF DEVELOPING SYNAPSES

描述（来自申请人摘要的逐字）：主要手段是 神经细胞之间的交流是通过释放 神经递质的化学突触。大脑处理信息的能力 信息依赖于精确可靠的突触连接 数百种不同类型的神经元之间。本申请针对 对调节突触的过程的分子理解 阵我们已经创建了突触囊泡蛋白-GFP融合体， 这些在线虫C. elegans使我们能够在光水平上可视化突触前的专业化 在活体动物中的分辨率利用这些转基因动物，我们分离出 突变体C.它们的形成能力受到了影响 神经元间突触特化。我们建议进一步分析这些 突变体，特别是集中在超微结构表征 与遗传病变相关的异常。其次，我们建议 这些突变体中的几个中受损的基因的分子克隆。分子 克隆这些基因中的第一个表明它编码一个大的蛋白质， 与脊椎动物脑中表达的蛋白质同源。第三，我们建议 鉴定的基因产物的详细分子遗传特征 通过我们的遗传分析来评估它们在突触形成过程中的作用， 发展而C.优雅线虫的复杂性可能比 脊椎动物，分析分子参与的过程中C。 优雅的人应该建立一个基本的框架， 与更普遍和可能保守的突触原则有关， 发展

DESCRIPTION (Verbatim From the Applicant's Abstract): The primary means by which nerve cells communicate with each other is through the release of neurotransmitter at chemical synapses. The ability of the brain to process information depends on synaptic connections forming precisely and reliably between hundreds of different types of neurons. This application is directed towards developing a molecular understanding of processes that regulate synapse formation. We have created synaptic vesicle protein-GFP fusions and expressed these in subsets of glutamatergic mechanosensory neurons in the nematode C. elegans enabling us to visualize presynaptic specializations at light level resolution in live animals. Using these transgenic animals, we have isolated mutants of C. elegans that are compromised in their ability to form interneuronal synaptic specializations. We propose to further analyze these mutants, particularly concentrating on ultrastructural characterization of abnormalities associated with the genetic lesions. Secondly, we propose molecular cloning of the genes lesioned in several of these mutants. Molecular cloning of the first of these genes suggests it encodes a large protein with homology to a protein expressed in the vertebrate brain. Thirdly, we propose the detailed molecular genetic characterization of the gene products identified through our genetic analyses to assess their roles in the process of synapse development. While synaptogenesis in C. elegans likely is less complex than in vertebrates, analysis of the molecules participating in the process in C. elegans should lay a basic scaffold on which to integrate the complexities associated with more general and likely conserved principles of synaptic development.