Identification and modulation of insula to BNST circuit specific afferents in ethanol abstinence

项目摘要 酒精使用障碍（AUD）是影响数百万美国人的主要健康问题， 成本超过2000亿美元。限制AUD影响的一个关键组成部分是防止复发， 酗酒者的共同苦难。压力极大地增加了恢复的可能性， 长期禁欲，AUD患者通常患有共病焦虑症。调制 内源性大麻素（eCB）系统可以降低对应激源的反应。英国广播公司和英国国家广播公司已经出现 作为涉及AUD停药阶段的区域。有证据表明，大脑皮层在以下方面起着至关重要的作用： 在禁欲期间经历的负性内感受性刺激。BNST是延伸杏仁核的一部分， 在成瘾中整合了奖励和压力我们的实验室已经发现了功能性单突触连接， BNST中的促肾上腺皮质激素释放因子（CRF）细胞（BNSTCRF细胞）。的 控制BNST投射岛（insulaBNST）神经元的输入和这些输入的调制仍然存在 重要的未知数这项研究旨在了解insulaBNST电路的调节以及如何 它在乙醇提取中发生变化。为此，我们将进一步研究传入神经的分布， insulaBNST细胞（子目标1.1），在传入区域之间建立单突触连接（子目标1.2），以及 询问乙醇和eCB在这些突触处功能（子目标1.3）。Subaim 2.1将确定 酒精戒断对该回路中离体切片生理学的影响，而子目标2.2将确定 通过纤维光度法使用钙成像的酒精戒断在体内的影响。我们希望调查结果 从这些实验中将提供一个更好的理解这些大脑区域之间的相互作用， 这种相互作用是如何调节的。这样做，我们将能够建立正常的生理和影响， 酒精戒断在这个新的电路上。

PROJECT SUMMARY Alcohol use disorder (AUD) is a major health concern affecting millions of Americans with an estimated annual cost of well over $200 Billion. A critical component in limiting the impact of AUD is preventing relapse, a common tribulation for alcoholics. Stress dramatically increases the likelihood of reinstatement during protracted abstinence, and patients with AUD often have comorbid anxiety disorders. Modulation of the endocannabinoid (eCB) system can decrease the response to stressors. The insula and BNST have emerged as regions implicated in the withdrawal phase of AUD. Evidence suggests that the insula plays a vital role in the negative interoceptive stimuli experienced during abstinence. The BNST, part of the extended amygdala, integrates reward and stress in addiction. Our lab has found functional monosynaptic connections between the insula and anxiety-responsive corticotrophin-releasing factor (CRF) cells in the BNST (BNSTCRF cells). The inputs that control the BNST projecting insular (insulaBNST) neurons and the modulation of those inputs remain essential unknowns. The proposed research aims to understand the regulation of the insulaBNST circuit and how it is changed in ethanol withdrawal. To do so, we will further investigate the distribution of afferents to insulaBNST cells (subaim 1.1), establish monosynaptic connectivity between afferent regions (subaim 1.2), and interrogate ethanol and eCB functionality at these synapses (subaim 1.3). Subaim 2.1 will ascertain the effects of alcohol withdrawal on ex vivo slice physiology in this circuit while subaim 2.2 will establish the implications of alcohol withdrawal in vivo using calcium imaging via fiber photometry. We hope the findings from these experiments will provide a better understanding of the interplay between these brain regions and how this interaction is regulated. In so doing, we will be able to establish the normal physiology and impact that alcohol withdrawal has on this novel circuit.