The role of presynaptic C1QL3 in a hypothalamic arousal circuit

项目概要： 发作性睡病与cataeprazole是一种衰弱的神经系统疾病，导致睡眠发作在白天， 晚上断断续续的睡眠。这种情况与下丘脑泌素/食欲素（HCRT/OX）神经元的丢失有关， 下丘脑外侧区（LHA），其是睡眠/觉醒行为和唤醒状态的重要调节器。 尽管有大量研究HCRT/OX神经元在生理和行为方面功能的工作， 但对其突触结构和功能的分子机制知之甚少。基于 根据我们实验室最近对LHA的单细胞RNA测序（scRNA-seq）分析的结果，我们提出C1 q13， 编码C1 QL 3，一种先前描述的在其他脑区的兴奋性突触组织蛋白， 同样是HCRT/OX神经元兴奋性突触传递的重要元素，并帮助我们摆脱 这些重要的唤醒调节突触的分子组成部分。为了验证这一点，我将确认 HCRT/OX神经元中C1 ql 3及其基因产物C1 QL 3在mRNA和蛋白质水平的表达 (Aim 1）。然后，我将有条件地敲除HCRT/OX神经元中的C1 ql 3基因，以确定结果 其缺失对HCRT/OX突触结构和功能的影响（目的2），以及其对 HCRT/OX神经元在促进觉醒中的功能（目的3）。这项工作将产生若干重要成果， 包括1）确定HCRT/OX神经元中新遗传标记的作用，2）告知我们 C1 QL 3在与HCRT/OX神经元相关的突触功能和行为中的作用，以及3）为我提供以下方面的培训： 神经科学技术，从细胞和分子到行为，使我成为一个更好的， 在我追求成为一名独立研究人员的目标时，我是一名全面的科学家和科学传播者。

PROJECT SUMMARY: Narcolepsy with cataplexy is a debilitating neurological disorder that results in sleep attacks during the day and fragmented sleep at night. This condition is associated with the loss of hypocretin/orexin (HCRT/OX) neurons in the lateral hypothalamic area (LHA), which are important regulators of sleep/wake behavior and arousal states. Despite a broad array of work investigating HCRT/OX neuron function in physiology and behavior, comparatively little is known about the molecular mechanisms underlying their synaptic structure and function. Based on the results of our lab’s recent single cell RNA sequencing (scRNA-seq) analysis of the LHA, we propose that C1ql3, encoding C1QL3, a previously described excitatory synapse organizing protein in other brain regions, may likewise be an important element in excitatory synaptic transmission in HCRT/OX neurons, and help us shed light on the molecular components of these important arousal-regulating synapses. To test this, I will confirm the expression of C1ql3 and its gene product C1QL3 in HCRT/OX neurons, at both the mRNA and protein levels (Aim 1). I will then conditionally knock out the C1ql3 gene in HCRT/OX neurons to determine the consequences of its deletion on HCRT/OX synaptic structure and function (Aim 2), as well as its potential importance for HCRT/OX neuron function in promoting arousal (Aim 3). This work will have several important outcomes, including 1) identifying the role of a novel genetic marker in HCRT/OX neurons, 2) informing us of the importance of C1QL3 in synaptic function and behavior relating to HCRT/OX neurons, and 3) provide me with training in neuroscience techniques ranging from cellular and molecular to behavioral to enable me to become a more well- rounded scientist and scientific communicator, as I pursue my goal of becoming an independent researcher.