Delineating the synapse coordination pathway

项目总结 大脑的所有功能，从感觉到行为，都源于突触的模式和特性 在数十亿个神经元之间发现了联系。这些突触的形式多种多样，但有两种 主要有两类：电突触和化学突触。虽然这些类被认为是不同的 生化单位，很明显这两类的发展是有联系的。例如，电力供应中断 突触改变化学突触的发育，导致各种行为缺陷，同时扰乱 化学突触影响电突触的形态和功能。田野中的一道屏障仍然存在 了解神经元如何协调这些不同类型突触的发展。引人注目的是，正在出现 有证据表明，这种突触协调可能是通过一种协调的调节途径实现的。我们的 研究发现与自闭症和癫痫相关的基因Neurobeachin对电气和化学都是必需的 通过结合两种类型突触的细胞内支架形成突触，这是 构建功能性突触。此外，我们的初步工作表明，神经导航素的干扰- 相互作用的分子也会导致电和化学突触形成的缺陷。这些碎片 证据开始建立一个模型，在这个模型中，突触协调是通过一条调控途径实现的，即 对于两种突触类型都是必不可少的。然而，在理解基因的全部范围方面仍然存在一个关键的差距 构成突触协调途径的调控网络。因此，这个项目的目标是 用遗传学方法确定调节体内电和化学突触协调的基因 无障碍脊椎动物模型系统。这项提案使用了斑马鱼莫特纳赛道和它的刻板印象 电和化学突触以及无与伦比的基因和成像可获得性来评估突触 脊椎动物体内的协调性。在Aim1中，我们建议确定协调电信号的候选基因 和化学突触的形成使用我们高效的CRISPR筛选管道。在AIM2中，我们研究 候选突变对电突触和化学突触形成和功能的影响。团结在一起， 拟议的研究将确定协调脑内电和化学突触形成所需的基因 活着。这项工作有可能揭示神经科学的一个新前沿，一条协调 电和化学突触的发育。

PROJECT SUMMARY All of brain function, from sensory perception to behavior, is derived from the pattern and properties of synaptic connections found between billions of individual neurons. These synapses are found in diverse forms, but two major classes exist: electrical and chemical synapses. While these classes are thought to be distinct biochemical units, it is clear the development of both types is linked. For example, the disruption of electrical synapses alters chemical synapse development resulting in a variety of behavioral defects, while perturbing chemical synapses impacts electrical synapse form and function. A barrier in the field remains in understanding how neurons coordinate the development of these distinct synapse types. Strikingly, emerging evidence suggests that such synaptic coordination may be achieved by a concerted regulatory pathway. Our work identifies the autism- and epilepsy-linked gene Neurobeachin as required for both electrical and chemical synapse formation by binding the intracellular scaffolds of both types of synapses, which are necessary for building functional synapses. Additionally, our preliminary work reveals that the disruption of Neurobeachin- interacting molecules also results in defects in both electrical and chemical synapse formation. These pieces of evidence begin to build a model wherein synapse coordination is achieved by a regulatory pathway that is essential for both synapse types. Yet, a critical gap remains in understanding the full extent of the genetic regulatory network that constitutes the synapse coordination pathway. Therefore, the goal of this project is to identify the genes that regulate electrical and chemical synapse coordination in vivo using a genetically accessible vertebrate model system. This proposal uses the zebrafish Mauthner circuit with its stereotyped electrical and chemical synapses and unparalleled genetic and imaging accessibility to assess synaptic coordination in vertebrates in vivo. In Aim1 we propose to identify candidate genes that coordinate electrical and chemical synapse formation using our efficient CRISPR screening pipeline. In Aim2 we examine the effects of candidate mutations on the formation and function of electrical and chemical synapses. Together, the proposed studies will identify the genes required to coordinate electrical and chemical synapse formation in vivo. This work has the potential to reveal a novel frontier in neuroscience, a pathway that coordinates electrical and chemical synapse development.