Core C - Arrigoni

项目总结/摘要 电生理学核心将为电生理学提供集中服务， 研究，包括体外脑切片的制备，膜片钳记录， 免疫组化及相关数据分析。项目1（Saper）、项目2 （Scammell）和项目4（Chamberlin）将使用该核心设施。我们将使用补丁- 钳位记录和通道视紫红质2（ChR 2）辅助电路映射（CRACM）， 对于项目1，确定CGRP表达之间的功能性突触连接性， 臂旁外侧核（PBel）神经元和基底节神经元 前脑（BF）、外侧下丘脑（LH）、乳头体上核（SUM）和中央 杏仁核（CeA）。对于项目2，我们将确定功能性突触连接 在背外侧和背外侧的胆碱能神经元、多巴胺能神经元和GABA能神经元之间， 脚桥被盖核（LDT/PPT）和BF、中线丘脑的神经元 和SUM，以及对于项目4， 舌下神经运动神经元。在这些实验中，我们 定义参与这些输入的神经递质和突触后受体。在 此外，核心将继续验证和优化刺激措施 光遗传学和药物遗传学工具的范例（ChR 2和ArchT兴奋性 和抑制性DREADD）。这些项目将受益于 核心的专业知识和共享资源。

Project Summary/Abstract The Electrophysiology Core will provide centralized services for electrophysiological studies, including preparation of in vitro brain slices, patch-clamp recordings, immunohistochemistry, and the associated data analysis. Project 1 (Saper), Project 2 (Scammell) and Project 4 (Chamberlin) will use this Core facility. We will use patch- clamp recordings and channelrhodopsin2 (ChR2)-assisted circuit mapping (CRACM) to determine, for Project 1, functional synaptic connectivity between CGRP expressing neurons of the external lateral parabrachial nucleus (PBel) and neurons of the basal forebrain (BF), lateral hypothalamus (LH), supramammillary nucleus (SUM) and central amygdala (CeA). For Project 2 we will determine functional synaptic connectivity between cholinergic, glutamatergic and GABAergic neurons of the laterodorsal and pedunculopontine tegmental nuclei (LDT/PPT) and neurons of the BF, midline thalamus and SUM, and for Project 4 between glutamatergic premotorneurons of the parahypoglossal zone and hypoglossal motoneurons. In these experiments we will define the neurotransmitters and postsynaptic receptors involved in these inputs. In addition, the Core will be continuing the validation and optimization of the stimulation paradigms for the opto-genetic and pharmaco-genetic tools (ChR2 and ArchT excitatory and inhibitory DREADDs) used by these Projects. The Projects will benefit from the expertise and shared resources of the Core.