In Vivo Time-Lapse Imaging of Retinal Synaptogenesis

描述(申请人提供)：高度具体和可重复性的图案 在中枢神经系统发育过程中建立了神经元的连接 神经系统(CNS)。单个神经元重塑其神经细胞结构 根据电活动的相对水平。大多数，如果不是全部的话，神经学的 这些“突触产生”过程的中断导致了疾病的发生。因为 神经元突触的形成是一个高度动态的过程，能够观察 这些事件发生时--最好是在完整的神经系统中--将 大大加强努力，阐明负责监管的机制 突触发生和突触调节。这项提议将在体内利用 时间推移成像研究细胞和分子机制 调节突触接触，组织成功能不同的板层， 在斑马鱼视网膜的发育过程中。这个系统有很多优点： (I)视网膜内的突触板层特征良好；(Ii)鱼 视网膜发育迅速(Iii)眼睛是透明的，因此非常适合 实时成像，(Iv)转基因和分子操纵技术 已建立的(V)转基因斑马鱼品系，在这些品系中存在视网膜亚群 神经元表达荧光标记蛋白，以及(Vi)突变鱼，其中 视网膜发育和/或神经元分层受到干扰是可能的。这个 这项研究的结果将为今后旨在 探讨在分子水平上组织突触发育的机制， 蜂窝和网络级别。

DESCRIPTION (provided by applicant): Highly specific and reproducible patterns of neuronal connectivity are established during development of the central nervous system (CNS). Individual neurons remodel their neuritic architecture based on relative levels of electrical activity. Most, if not all, neurological disorders result from disruption of these "synaptogenic" processes. Because neuronal synapse formation is a highly dynamic process, the ability to watch these events as they occur - preferably in the intact nervous system - will greatly enhance efforts to elucidate the mechanisms responsible for regulating both synaptogenesis and synaptic modulation. This proposal will utilize in vivo time-lapse imaging to investigate the cellular and molecular mechanisms that regulate synaptic contacts, organized into functionally distinct laminae, during development of the zebrafish retina. This system has many advantages: (i) synaptic laminae within the retina are well-characterized, (ii) the fish retina develops rapidly, (iii) the eye is transparent and therefore ideal for live imaging, (iv) transgenic and molecular manipulation techniques are established, (v) transgenic zebrafish lines exist in which subsets of retinal neurons express fluorescent marker proteins, and (vi) mutant fish in which retinal development and/or neuronal lamination is perturbed are available. The results of this study will provide the foundation for future studies aimed at addressing the mechanisms that organize synaptic development at the molecular, cellular and network levels.