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 尽管已广泛报道了酒精使用障碍（AUD）对谷氨酸能信号传导的影响，但该作用 AUD中的三角洲谷氨酸受体（GLUD1和GLUD2）的含量尚不清楚。尽管他们没有参与 众所周知，GLUD家族对谷氨酸的传统兴奋性突触反应进行了调节 兴奋性信号传导可能导致在AUD模型中观察到的神经元活性功能障碍。 GLUD1也与几项全基因组关联研究有关。但是，没有研究 已直接评估酒精暴露对GLUD1功能的影响。 脊柱末端（BNST）的床核是一种前脑核，已被严重牵涉到关键 酒精暴露后神经调节的枢纽。先前的工作表明酒精暴露会调节 BNST中的突触谷氨酸，但这种情况的确切机制尚不清楚。突触 谷氨酸能传播部分是由AMPA受体介导的，其中约30％可以通过钙（钙 - 可渗透的Ampars，CP-Ampars）。我们的初步数据表明，急性退出慢性 间歇性乙醇蒸气暴露（CIE）降低了CP-Ampar突触电流并降低了GLUD1 BNST中介导的补他的兴奋剂。尽管已经在表达Glud1之前观察到了 BNST，这些是第一个数据可能参与调节BNST信号的数据。这些数据也是如此 暗示GLUD1调制是酒精暴露后神经调节的可能新靶标。这 建议旨在进一步研究CP-Ampars和Glud1在BNST中的作用，它们如何相互作用以及 它们如何使用荧光原位杂交，斑块钳的结合使用酒精暴露。 电生理学和离体钙成像。 AIM 1将使用Glud1敲除（GLUD1 KO）小鼠和野生型 （wt）同窝仔在BNST中建立Glud1的功能曲线，并与CP-Ampar重叠 表达BNST细胞。然后，AIM 2将评估CIE对BNST CP-AMPAR和GLUD1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。最后，目标3 将使用这些方法来检查BNST CP-AMPAR和GLUD1信号的转化相关性 跨不同型号的AUD。