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

项目摘要/摘要 戒酒会导致消极的情绪状态，从而导致不良适应反应 压力和故态复萌。临床前研究已经开始识别神经回路和调节 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 参与上瘾和消极情绪，下一步是了解这些人是如何进行沟通的 神经回路尤其在神经肽和微回路水平上受到调节。纹状体的床核 终末(BNST)是戒断相关神经回路的基本组成部分，因为它调节应激和 以神经肽依赖的方式与酒精相关的行为。研究多肽特异的BNST回路 调控戒断时的负性情绪，我们将重点关注臂旁核(PBN)的传入， 脑干区域起危险信号作用的脑干区域PBN向BNST释放降钙素基因的投射 相关肽(CGRP)和垂体腺苷环化酶激活多肽(PACAP)，这些肽 分别调节疼痛和恐惧回路。我们的研究涉及异性型和性二型性 当PBN投射诱导BNST细胞的异质性体外活动时，在PBNàBNST回路中进行控制 以及BNSTPBN激活的女性特有的焦虑样行为。此外，我们的初步研究 建议PBN在酒精戒断中发挥作用，因为酒精暴露后的失活是缓解焦虑的。这 提案将通过调查CGRP和PACAP如何 参与PBNàBNST环路诱导的戒断行为，在体内和体外均有活性。 因此，指导K99阶段将建立在我的体内纤维光度记录和提供培训 在体外记录中确定降钙素基因相关肽对BNSTàPBN活性、戒断诱导行为的作用 (AIM1)和PACAP在CGRP神经调节中的作用(AIM2)。独立的R00阶段 将在微电路水平和酒精相关状态下研究PACAP对BNSTáPBN的作用，并与 目的进一步阐明多肽串扰(AIM3)的复杂性。拟修读的课程及相关职业 发展培训计划在这条通往独立之路的马赛克奖集体提供理想 将申请者转变为独立成瘾神经学家的机制。结果将会是 极大地促进了我们在多肽和微电路水平上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.