Development of a model system for presynaptic study

描述(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涉及带状视网膜神经元神经递质释放的分子的系统。斑马鱼是研究生理和疾病的优秀脊椎动物模型，既有利于大规模的突变筛选，也有利于快速廉价地产生转基因动物。因此，已经产生了几个作为人类疾病模型的突变体，未来还将开发更多的突变体。我们打算将这一准备工作扩展到研究视觉建立后带状突触的突触前终末的生理学。这项建议旨在开发工具和技术来操纵和研究这些细胞的突触前过程。目的1建立两种斑马鱼制剂，用于研究视网膜神经元的神经递质释放和囊泡回收。目的建立几个转基因细胞系，用于监测视网膜神经元的胞吐和胞吞特性。目的3试图开发新的工具来沉默成年斑马鱼的基因表达。神经系统中的信息在突触的神经细胞之间传输，突触前神经细胞中的电脉冲导致神经递质从被膜结合的小结构释放，称为突触小泡。这一过程中的缺陷被认为是许多神经疾病的基础和病理因素，包括亨廷顿病、阿尔茨海默病、双相情感障碍、精神分裂症、帕金森病和智力低下，但这些疾病如何影响突触前功能尚不清楚。这项提议开发了新的工具来研究突触前神经细胞如何释放神经递质。

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.