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: 8482355
• 负责人: REBECCA P SEAL
• 金额: $ 36.38万
• 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/8482355
• 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); 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; 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; improved; in vivo; insight; nervous system disorder; neuropsychiatry; new therapeutic target; novel; patch clamp; public health relevance; recombinase; 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脑功能的许多方面具有关键作用，包括学习和记忆、注意力、奖赏和运动功能，并且它们的失调导致神经精神障碍，例如帕金森病、精神分裂症和阿尔茨海默病。几十年来，人们认为这些细胞只释放一种经典的递质乙酰胆碱，但现在知道它们表达囊泡谷氨酸转运蛋白3并释放谷氨酸。虽然我们有数据表明，VGLUT 3在纹状体胆碱能神经元介导快速的神经递质传递和调节胆碱能传递，不太知道它在基底前脑神经元的作用或其行为的作用，无论是人口。因此，该基金的目的是确定VGLUT 3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VGLUT 3在基底前脑核的细胞分布，并使用我们的BAC转基因cre重组酶和报告小鼠连同cre依赖的病毒追踪方法绘制其轴突投射。这些新工具提供了比传统方法更高的灵敏度和选择性。在目标2中，我们将确定VGLUT 3介导的突触信号传导的基底前脑胆碱能神经元的机制。我们将使用生物化学方法来评估VGLUT 3和VAChT是否驻留在相同的囊泡上，这表明协同共包装增加了这些细胞的胆碱能传递。我们将使用视紫红质通道依赖性病毒表达与膜片钳电生理学一起检测胆碱能细胞的快速多巴胺能传递。在目标3中，我们将确定VGLUT 3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VGLUT 3敲除小鼠，我们现在有证据表明胆碱能神经元中的行为与VGLUT 3明确相关。这些神经元中的转运蛋白的缺失产生夜间运动过度活动以及两个工作记忆任务中的缺陷，新的手臂识别和自发交替。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。靶向前脑胆碱能神经元的信号传导的促胆碱药通常用于治疗许多神经系统疾病。我们的工作表明，这些细胞的谷氨酸信号可以提供新的治疗靶点。