Role of Cholinergic-Glutamatergic Co-transmission in Forebrain Circuits

胆碱能-谷氨酸共同传输在前脑回路中的作用

• 批准号: 9231506
• 负责人: REBECCA P SEAL
• 金额: $ 34.45万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9231506
• 关键词: Acetylcholine; Alzheimer' s Disease; Attention; Behavior; Behavioral; Biochemical; Brain; Cell Nucleus; Cells; Cognition; Cognitive; Corpus striatum structure; Data; Defect; Disease; Dopamine; Electrophysiology (science); Enterobacteria phage P1 Cre recombinase; Genetic; Glutamate Transporter; Glutamates; Goals; Grant; Hyperactive behavior; Knockout Mice; Learning; Link; Literature; Location; Maps; Mediating; Memory; Mental disorders; Methods; Moods; Motor; Mus; Neurologic Dysfunctions; Neurons; Neurotransmitters; Nicotine; Parkinson Disease; Phenotype; Population; Prosencephalon; Regulation; Reporter; Resolution; Rewards; Role; SLC17A8 gene; Schizophrenia; Short-Term Memory; Signal Transduction; Slice; Synapses; Techniques; Transgenic Organisms; Vesicle; Viral; Work; acetylcholine transporter; arm; basal forebrain; basal forebrain cholinergic neurons; cholinergic; cholinergic neuron; glutamatergic signaling; improved; in vivo; insight; nervous system disorder; neuropsychiatric disorder; new therapeutic target; novel; patch clamp; public health relevance; quantum; symporter; tool; transmission process

DESCRIPTION (provided by applicant): Basal forebrain and striatal cholinergic neurons have a critical role in many aspects of brain function including learning and memory, attention, reward and motor function and their dysregulation contributes to neuropsychiatric disorders such as Parkinson's disease, schizophrenia and Alzheimer's disease. For decades, it was assumed that these cells release only one classical transmitter, acetylcholine, however it is now known that they express the vesicular glutamate transporter 3 and release glutamate. While we have data showing that VGLUT3 in striatal cholinergic neurons mediates fast glutamatergic transmission and modulates cholinergic transmission, not much is known about its role in basal forebrain neurons or its behavioral role in either population. Thus, the goal of this grant is to determine te functional and behavioral role of VGLUT3 in forebrain cholinergic neurons. In Aim 1 we will determine the cellular distribution of VGLUT3 in basal forebrain nuclei and map their axonal projections using our BAC transgenic cre recombinase and reporter mice together with cre-dependent viral tracing methods. These new tools provide higher sensitivity and selectivity over more traditional methods. In Aim 2 we will determine mechanisms of VGLUT3-mediated synaptic signaling by basal forebrain cholinergic neurons. We will use a biochemical approach to assess whether VGLUT3 and VAChT reside on the same vesicles, which would suggest synergistic co-packaging increases cholinergic transmission by those cells. We will use cre-dependent viral expression of channelrhodopsin together with patch clamp electrophysiology to detect fast glutamatergic transmission by cholinergic cells. In Aim 3 we will determine the behavioral role of VGLUT3 in striatal and basal forebrain cholinergic neurons. Using our conditional VGLUT3 knockout mice, we now have evidence of behaviors definitively linked to VGLUT3 in cholinergic neurons. Deletion of the transporter in these neurons produces nocturnal hyperlocomotor activity as well as deficits in two working memory tasks, novel arm recognition and spontaneous alternation. Work in this aim will determine which population of cholinergic neurons mediates these phenotypes and if there is a behavioral role for the glutamate signaling by these cells. Cholinomemetics that target signaling by forebrain cholinergic neurons are commonly used to treat a number of neurological disorders. Our work here suggests that glutamate signaling by these cells could provide novel therapeutic targets.

描述（由申请人提供）：基底前脑和纹状体胆碱能神经元在大脑功能的许多方面发挥着关键作用，包括学习和记忆、注意力、奖励和运动功能，它们的失调会导致帕金森病、精神分裂症和阿尔茨海默病等神经精神疾病。几十年来，人们一直认为这些细胞只释放一种经典递质，即乙酰胆碱，但现在已知它们表达囊泡谷氨酸转运蛋白 3 并释放谷氨酸。虽然我们有数据显示纹状体胆碱能神经元中的 VGLUT3 介导快速谷氨酸能传递并调节胆碱能传递，但对其在基底前脑神经元中的作用或其在任一群体中的行为作用知之甚少。因此，这项资助的目标是确定 VGLUT3 在前脑胆碱能神经元中的功能和行为作用。在目标 1 中，我们将确定 VGLUT3 在基底前脑核中的细胞分布，并使用我们的 BAC 转基因 cre 重组酶和报告小鼠以及依赖于 cre 的病毒追踪方法来绘制其轴突投影图。这些新工具比传统方法具有更高的灵敏度和选择性。在目标 2 中，我们将确定基底前脑胆碱能神经元 VGLUT3 介导的突触信号传导机制。我们将使用生化方法来评估 VGLUT3 和 VAChT 是否驻留在同一囊泡上，这表明协同共包装会增加这些细胞的胆碱能传递。我们将使用视紫红质通道的cre依赖性病毒表达以及膜片钳电生理学来检测胆碱能细胞的快速谷氨酸能传递。在目标 3 中，我们将确定 VGLUT3 在纹状体和基底前脑胆碱能神经元中的行为作用。使用我们的条件性 VGLUT3 基因敲除小鼠，我们现在有证据表明胆碱能神经元中的行为与 VGLUT3 明确相关。这些神经元中转运蛋白的缺失会导致夜间运动过度，以及两项工作记忆任务（新手臂识别和自发交替）的缺陷。这一目标的工作将确定哪些胆碱能神经元群体介导这些表型，以及这些细胞的谷氨酸信号传导是否具有行为作用。针对前脑胆碱能神经元信号传导的胆碱模拟剂通常用于治疗许多神经系统疾病。我们在这里的工作表明，这些细胞的谷氨酸信号传导可以提供新的治疗靶点。