Development of a model system for presynaptic study

描述（由申请人提供）：本项目的目标是建立斑马鱼双极细胞作为研究带状型视网膜神经元神经递质释放相关分子的系统。斑马鱼是研究生理学和疾病的一种极好的脊椎动物模型，有利于大规模诱变筛选和快速廉价地产生转基因动物。因此，已经产生了几种突变体，作为人类疾病模型，未来将开发更多的突变体。我们打算扩大这种制备的实用性，从带状突触后的视觉已经建立突触前末梢的生理学研究。该建议旨在开发工具和技术，用于操纵和研究这些细胞中的突触前过程。目的1建立两种研究视网膜神经元神经递质释放和囊泡再循环的斑马鱼标本。目的2建立几个转基因细胞系，用于检测视网膜神经元胞吐和胞吞的特性。目的3试图开发新的沉默成年斑马鱼基因表达的工具。神经系统中的信息在突触处的神经细胞之间传递，其中“突触前”神经细胞中的电脉冲导致神经递质从被称为突触囊泡的小膜结合结构释放。已经提出，该过程中的缺陷是许多神经障碍的基础并促成其病理，包括亨廷顿病、阿尔茨海默病、双相情感障碍、精神分裂症、帕金森病和智力迟钝，但尚不清楚这些障碍如何影响突触前功能。这一提议为研究突触前神经细胞如何释放神经递质提供了新的工具。

DESCRIPTION (provided by applicant): The goal of this project is to establish the zebrafish bipolar cells as a system for studying the molecules involved neurotransmitter release from ribbon-type retinal neurons. Zebrafish are an excellent vertebrate model for physiology and disease, conducive to both large-scale mutagenesis screens and the rapid and cheap generation of transgenic animals. Hence, several mutants have been generated that serve as human disease models and many more will be developed in the future. We intend to extend the utility of this preparation to the study of the physiology of presynaptic terminals from ribbon-type synapses after vision has been established. This proposal aims at developing tools and techniques for manipulation and investigation of presynaptic processes in these cells. Aim 1 establishes two zebrafish preparations for studying neurotransmitter release and vesicle recycling from retinal neurons. Aim 2 generates several transgenic lines for monitoring the properties of exocytosis and endocytosis in retinal neurons. Aim 3 attempts to develop new tools for silencing gene expression in adult zebrafish. Information in the nervous system is transmitted between nerve cells at the synapse, where an electrical impulse in the "presynaptic" nerve cell causes the release of neurotransmitter from small membrane bound structures, known as synaptic vesicles. Deficiencies in this process have been suggested to underlie and contribute to the pathologies of a number of neurological disorders, including Huntington's disease, Alzheimer's disease, bipolar disorder, Schizophrenia, Parkinson's Disease and mental retardation, yet it is unclear how presynaptic function is affected by these disorders. This proposal develops new tools to study how presynaptic nerve cells release neurotransmitter.