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) 对谷氨酸信号传导的影响已被广泛报道，但其作用 AUD 中 δ 谷氨酸受体（GluD1 和 GluD2）的作用仍然未知。虽然他们没有参与其中 与传统的谷氨酸兴奋性突触反应不同，GluD 家族可以介导许多更微妙的方面 兴奋性信号传导可能导致 AUD 模型中观察到的神经元活动功能障碍。 多项全基因组关联研究也表明 GluD1 与 AUD 相关。然而，尚无研究 直接评估酒精暴露对 GluD1 功能的影响。 终纹床核 (BNST) 是一个前脑核，被认为是一个重要的神经系统。 酒精暴露后的神经调节中心。先前的研究表明，酒精暴露可以调节 BNST 中的突触谷氨酸，但发生这种情况的确切机制仍不清楚。突触 谷氨酸能传递部分由 AMPA 受体介导，其中约 30% 可以传递钙（钙- 渗透性 AMPAR、CP-AMPAR）。我们的初步数据表明，从慢性病中急性戒断 间歇性乙醇蒸气暴露 (CIE) 会降低 CP-AMPAR 突触电流并降低 GluD1 BNST 介导的强直兴奋电流。尽管之前已经观察到GluD1表达于 BNST，这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 GluD1 调节可能是酒精暴露后神经调节的新靶标。这 该提案旨在进一步研究 CP-AMPAR 和 GluD1 在 BNST 中的作用、它们如何相互作用以及 使用荧光原位杂交、膜片钳组合来研究酒精暴露对它们的影响 电生理学和离体钙成像。目标 1 将使用 GluD1 敲除 (GluD1 KO) 小鼠和野生型 (WT) 同窝小鼠建立 BNST 中 GluD1 的功能图谱，以及它与 CP-AMPAR 的重叠 表达 BNST 细胞。然后目标 2 将评估 CIE 对 BNST CP-AMPAR 和 GluD1 表达的影响 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。最后，目标3 将使用这些方法来检查 BNST CP-AMPAR 和 GluD1 信号传导的翻译相关性 跨越不同模型的澳元。