The role of presynaptic C1QL3 in a hypothalamic arousal circuit

项目总结： 发作性睡病合并猝发是一种衰弱的神经疾病，会导致白天和 晚上睡得支离破碎。这种情况与下丘脑皮层下丘脑促性腺激素/食欲素(HCRT/OX)神经元的丢失有关 下丘脑外侧区(LHA)，它是睡眠/觉醒行为和觉醒状态的重要调节区域。 尽管对HCRT/OX神经元在生理和行为方面的功能进行了广泛的研究，但 对其突触结构和功能背后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C1q13， 编码C1QL3，一种先前在其他大脑区域描述的兴奋性突触组织蛋白，可能 同样，它也是HCRT/OX神经元兴奋性突触传递的重要因素，并帮助我们摆脱 了解这些重要的唤醒调节突触的分子组成。为了测试这一点，我将确认 C1qL3及其基因产物C1QL3在HCRT/OX神经元中的表达 (目标1)。然后，我将有条件地敲除HCRT/OX神经元中的C1ql3基因，以确定其后果 它在HCRT/OX突触结构和功能上的缺失(目标2)，以及它在 HCRT/OX神经元在促进觉醒中的作用(目标3)。这项工作将有几个重要的结果， 包括1)确定一种新的遗传标记在HCRT/OX神经元中的作用，2)告知我们其重要性 C1QL3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 神经科学技术，从细胞和分子到行为，使我成为一个更好的- 全能的科学家和科学传播者，因为我追求成为一名独立研究人员的目标。

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.