Neuropeptide-dependent parabrachial control of the BNST during alcohol abstinence-induced negative affect

项目总结/摘要 戒酒会诱发一种消极的情感状态，从而导致适应不良的反应 压力和复发。临床前研究已经开始确定神经回路和肽靶点， 禁欲期间的负面影响随着该领域开始对电路有了更深入的了解， 参与成瘾和负面影响，下一步是了解如何在这些沟通 特别是在神经肽和微回路水平上调节神经回路。纹床核 终末肌（BNST）是禁欲相关神经回路的基本组成部分，因为它调节压力， 神经肽依赖性的酒精相关行为。研究肽特异性BNST电路 在禁欲期间调节负面情绪，我们将重点关注臂旁核（PBN）的传入， 一个脑干区域，作为危险信号。PBN投射到BNST释放降钙素基因- 相关肽（CGRP）和垂体腺苷酸环化酶激活多肽（PACAP）， 分别调节疼痛和恐惧回路。我们的研究表明， PBN投射在BNST细胞中诱导异源离体活性时，PBNàBNST回路内的控制 和女性特有的焦虑样行为与BNSTPBN激活。此外，我们的初步研究 提示PBN在酒精戒断中的作用，因为酒精暴露后的失活是抗焦虑的。这 一项提案将通过调查CGRP和PACAP如何在很大程度上建立在这一基础证据的基础上， 有助于PBNàBNST回路诱导的戒断诱导行为、体内和离体活性。 因此，指导K99阶段将建立在我的体内纤维光度测定记录的基础上，并提供培训 在离体记录中，以确定CGRP对BNSTàPBN活性、戒断诱导行为 （AIM 1），以及PACAP对CGRP神经调节的贡献（AIM 2）。独立R 00阶段 将在微电路水平和酒精相关状态下研究PACAP对BNSTàPBN的作用， 目的是进一步描述肽串扰（AIM 3）的复杂性。建议的研究和相关职业 发展培训计划在这个马赛克独立之路奖集体提供了理想的 机制过渡到申请人的职业生涯作为一个独立的成瘾神经科学家。结果将 在肽和微回路水平上显著推进我们对神经回路机制的理解， 长期戒酒，同时告知在酒精使用障碍中使用肽能药物疗法。

PROJECT SUMMARY/ABSTRACT Abstinence from alcohol use induces a negative affective state that can lead to maladaptive responses to stress and relapse. Preclinical studies have begun to identify neurocircuits and peptide targets that regulate negative affect during abstinence. As the field has begun to develop a deeper understanding of the circuitries participating in addiction and negative affect, the next step is to understand how communication within these circuits is modulated particularly at the neuropeptide and microcircuit level. The bed nucleus of the stria terminalis (BNST) is a fundamental component of abstinence-relevant neurocircuitry as it modulates stress and alcohol-related behavior in a neuropeptide-dependent manner. To investigate peptide-specific BNST circuitry modulating negative-affect during abstinence, we will focus on afferents from the parabrachial nucleus (PBN), a brainstem region that functions as a danger signal. PBN projections to the BNST release calcitonin gene- related peptide (CGRP) and pituitary adenylate cyclase activating polypeptide (PACAP), peptides that modulate pain and fear circuits, respectively. Our studies implicate heterogenous and sexually dimorphic control within the PBNàBNST circuit as PBN projections induce heterogeneous ex vivo activity in BNST cells and female-specific anxiety-like behavior with BNSTPBN activation. Furthermore, our preliminary studies suggest a role for the PBN in alcohol-withdrawal, as inactivation is anxiolytic following alcohol exposure. This proposal will significantly build on this foundational evidence by investigating how CGRP and PACAP contribute to PBNàBNST circuit induced abstinence-induced behavior, in vivo and ex vivo activity. Accordingly, the mentored K99 phase will build on my in vivo fiber photometry recordings and provide training in ex vivo recordings to determine the role of CGRP on BNSTàPBN activity, abstinence-induced behavior (AIM1), and the contribution of PACAP on the CGRP-neuromodulation (AIM2). The independent R00 phase will investigate the role of PACAP on BNSTàPBN at the microcircuit level and alcohol-related states, with the goal to further delineate the intricacies of peptide crosstalk (AIM3). The proposed studies and related career development training plan in this MOSAIC Pathway to Independence Award collectively provide the ideal mechanism to transition the applicant to a career as an independent addiction neuroscientist. The results will significantly advance our understanding of neurocircuit mechanisms at the peptide and microcircuit level in protracted abstinence while informing the use of peptidergic pharmacotherapies in alcohol use disorders.