Investigating tonic and synaptic excitatory signaling in the bed nucleus of the stria terminalis across models of alcohol exposure

研究酒精暴露模型中终纹床核的强直和突触兴奋信号传导

• 批准号: 10825889
• 负责人: Sara Yi-Ling Conley
• 金额: $ 3.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-14 至 2025-11-27
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10825889
• 关键词: AMPA Receptors; Acute; Affect; Affective Symptoms; Alcohol consumption; Alcohols; Anterolateral; Anxiety; Area; Attenuated; Brain; Brain region; Calcium; Cell Nucleus; Cells; Chronic; Coupled; Data; Dimensions; Dorsal; Electrophysiology (science); Ethanol; Event; Excitatory Postsynaptic Potentials; Exposure to; Family; Female; Fluorescent in Situ Hybridization; Functional disorder; Genetic; Genotype; Glutamate Receptor; Glutamates; Goals; Health; Image; In Situ Hybridization; Knock-out; Knockout Mice; Knowledge; Ligands; Link; Long-Term Depression; Macaca; Macaca mulatta; Mediating; Messenger RNA; Modeling; Mus; Neuronal Dysfunction; Neurons; Pattern; Permeability; Pharmaceutical Preparations; Phenotype; Population; Prosencephalon; Reporting; Research; Role; Self Administration; Sex Differences; Signal Transduction; Stress; Structure of terminal stria nuclei of preoptic region; Synapses; Synaptic plasticity; System; Testing; Withdrawal; Work; addiction; alcohol effect; alcohol exposure; alcohol relapse; alcohol seeking behavior; alcohol use disorder; clinical development; clinically relevant; effective therapy; experimental study; genome wide association study; glutamatergic signaling; in vivo calcium imaging; insight; interest; mRNA Expression; male; mouse model; negative affect; neuronal excitability; neuroregulation; nonhuman primate; novel; patch clamp; receptor; response; sex; species difference; targeted treatment; transmission process; vapor

Abstract/Project Summary While the effects of alcohol use disorder (AUD) on glutamatergic signaling have been widely reported, the role of delta glutamate receptors (GluD1 and GluD2) in AUD remains unknown. Although they are not involved in traditional excitatory synaptic responses to glutamate, the GluD family is known to mediate many subtler aspects of excitatory signaling which may contribute to the dysfunctions of neuronal activity observed in models of AUD. GluD1 has also been implicated in AUD by several genome-wide association studies. However, no research has been conducted directly evaluating the effect of alcohol exposure on GluD1 function. The bed nucleus of the stria terminalis (BNST) is a forebrain nucleus that has been heavily implicated as a critical hub for neuromodulation following alcohol exposure. Previous work has shown that alcohol exposure regulates synaptic glutamate in the BNST, but the exact mechanisms by which this happen remain unclear. Synaptic glutamatergic transmission is in part mediated by AMPA receptors, ~30% of which can pass calcium (calcium- permeable AMPARs, CP-AMPARs). Our preliminary data suggests that acute withdrawal from chronic intermittent ethanol vapor exposure (CIE) decreases both CP-AMPAR synaptic current and reduces a GluD1 mediated tonic excitatory current in the BNST. Although it has been observed before that GluD1 is expressed in the BNST, these are the first data to that they may be involved in regulating BNST signaling. These data also implicate GluD1 modulation as a possible novel target for neuromodulation following alcohol exposure. This proposal aims to further examine the role of CP-AMPARs and GluD1 in the BNST, how they may interact, and how they are affected by alcohol exposure using a combination of fluorescent in situ hybridization, patch-clamp electrophysiology, and ex vivo calcium imaging. Aim 1 will use GluD1 knockout (GluD1 KO) mice and wildtype (WT) littermates to establish a functional profile of GluD1 in the BNST, as well as its overlap with CP-AMPAR expressing BNST cells. Aim 2 will then evaluate the impact of CIE on BNST CP-AMPAR and GluD1 expression and signaling, neuronal excitability, and spontaneous calcium dynamics in these mouse models. Finally, Aim 3 will use these approaches to examine the translational relevance of BNST CP-AMPAR and GluD1 signaling across different models of AUD.

摘要/项目摘要 虽然酒精使用障碍（AUD）对多巴胺能信号传导的影响已被广泛报道，但其作用 δ谷氨酸受体（GluD 1和GluD 2）在AUD中的作用尚不清楚。虽然他们没有参与 传统的兴奋性突触对谷氨酸的反应，已知GluD家族介导许多微妙的方面 兴奋性信号可能导致AUD模型中观察到的神经元活动功能障碍。 GluD 1也被几个全基因组关联研究牵连在AUD中。然而，没有研究表明 直接评估酒精暴露对GluD 1功能的影响。 终纹床核（BNST）是一个前脑核团，它被认为是一个重要的神经元。 酒精暴露后神经调节中枢。以前的研究表明，酒精暴露调节 突触谷氨酸在BNST，但这种情况发生的确切机制尚不清楚。突触 谷氨酸能传递部分由AMPA受体介导，约30%的AMPA受体可以传递钙（钙- 可渗透的AMPAR，CP-AMPAR）。我们的初步数据表明，急性戒断从慢性 间歇性乙醇蒸汽暴露（CIE）降低CP-AMPAR突触电流和减少GluD 1 介导的紧张性兴奋电流在BNST。虽然之前已经观察到GluD 1在大肠杆菌中表达， BNST，这些是它们可能参与调节BNST信号传导的第一个数据。这些数据还 暗示GluD 1调节是酒精暴露后神经调节可能的新靶点。这 该提案旨在进一步研究CP-AMPAR和GluD 1在BNST中的作用，它们如何相互作用， 使用荧光原位杂交，膜片钳， 电生理学和离体钙成像。目的1将使用GluD 1敲除（GluD 1 KO）小鼠和野生型 (WT)建立BNST中GluD 1的功能谱，以及其与CP-AMPAR的重叠 表达BNST细胞。目的2然后将评估CIE对BNST CP-AMPAR和GluD 1表达的影响 以及这些小鼠模型中的信号传导、神经元兴奋性和自发钙动力学。第三，目标 我将使用这些方法来研究BNST CP-AMPAR和GluD 1信号转导的翻译相关性 在不同的AUD模型中。