Mechanisms of Presynaptic Maintenance in C. elegans

项目摘要 虽然越来越多的工作有助于我们对Synapse的理解 对于突触如何在整个生命周期中保持不变，人们知之甚少。这漫长的一生 突触对神经回路的持续功能至关重要，包括那些支持 认知、运动和其他重要功能。然而，维持长寿命的突触 连接是一个细胞生物学挑战，因为突触蛋白的寿命是有限的， 突触小泡快速转换，突触前的蛋白质合成很少 车厢。本项目将研究突触前维持的机制。 雅致作为一种模范系统。在目标1中，我将研究与维持 突触结构，使用生长素诱导的降解系统来去除候选蛋白 来自成熟神经系统的Syd-2、Syd-1、SAD-1、CDK-5和PCT-1。中的更改 将使用突触的内源性细胞类型特异性标记物来评估突触组织 囊泡和活动区蛋白。在目标2中，我将确定突触前的调节者 支架蛋白SYD-2。我将实施一个可视的向前遗传屏幕来识别候选人 其调节SYD-2稳定性，使用脉冲追逐SYD-2 HaloTag方法可视化SYD-2 营业额。在目标3中，我将通过使用Split-TurboID近似性来识别SYD-2的调节器 生物素化以检测突触前间隔中SYD-2的相互作用伙伴。侯选人 SYD-2周转调节器将使用SYD-2 HaloTag系统进行评估。这项工作将 确定关键的突触维持蛋白及其调节因子。了解相关机制 保持稳定的突触为在衰老过程中保存突触提供了治疗途径 和神经退行性疾病。这个项目是在沈康博士的实验室里进行的 斯坦福大学为我提供了一个很好的细胞生物学和 神经科学，我将获得线虫模型系统的新经验，基因 操控和显微镜。

Project Summary While a growing body of work has contributed to our understanding of synapse assembly, less is known about how synapses are maintained throughout life. This long life of synapses is crucial for the sustained function of neural circuits, including those supporting cognition, movement, and other vital functions. However, maintaining long-lived synaptic connections presents a cell biological challenge, as synaptic proteins have finite lifetimes, synaptic vesicles turnover rapidly, and protein synthesis is scarce in the presynaptic compartment. This project will study the mechanisms of presynaptic maintenance, using C. elegans as a model system. In Aim 1, I will investigate the proteins involved in maintaining synaptic structures, using the auxin-inducible degron system to remove the candidate proteins SYD-2, SYD-1, SAD-1, CDK-5, and PCT-1 from the mature nervous system. Changes in synapse organization will be assessed using endogenous, cell-type specific markers of synaptic vesicles and active zone proteins. In Aim 2, I will identify regulators of the presynaptic scaffolding protein SYD-2. I will implement a visual forward genetic screen to identify candidates that regulate SYD-2 stability, using a pulse-chase SYD-2 HaloTag approach to visualize SYD-2 turnover. In Aim 3, I will identify regulators of SYD-2 through the use of Split-TurboID proximity biotinylation to detect interacting partners of SYD-2 in the presynaptic compartment. Candidate regulators of SYD-2 turnover will be assessed using the SYD-2 HaloTag system. This work will identify key synaptic maintenance proteins and their regulators. Understanding the mechanisms that maintain stable synapses provides therapeutic avenues for preserving synapses in aging and neurodegenerative diseases. This project, performed in the laboratory of Dr. Kang Shen at Stanford University, provides a strong training opportunity for me in the fields of cell biology and neuroscience, and I will gain new experience with the C. elegans model system, genetic manipulations, and microscopy.