The role of medial prefrontal cortex CRF signaling in binge-like ethanol intake

项目摘要/摘要 酒精依赖是一个广泛的公共卫生问题，可供选择的治疗方法有限。一个不断增长的 大量证据支持过量摄入乙醇是乙醇发展的一个重要风险因素 依赖。此外，这种形式的过量酒精摄入与负面健康影响有关，如 与心脏病和糖尿病一样，并占酒精造成的经济负担的很大一部分 在美国境内使用。我们的实验室和其他实验室已经广泛展示了神经肽促肾上腺皮质激素 释放因子(CRF)在调节酒精暴饮量中起着重要作用。更进一步地，改变 已知CRF系统在乙醇相关行为中发挥重要作用 从酒精依赖到戒断。CRF在狂饮行为中的具体作用 因此，不依赖酒精的动物能够发现这种危险行为模式的治疗方法。 此外，还提供了对与乙醇相关的系统的初始更改的重要见解 酒精依赖的发展和维持。促肾上腺皮质激素释放激素在酒精相关行为中的作用 延伸的杏仁核一直是众多研究的焦点。然而，CRF在区域内的作用 对于控制这些边缘区域和协调奖赏导向/抑制不当行为至关重要 从根本上被忽视了。这项建议的重点是通过审查来解决文献中的这个漏洞 内侧前额叶皮质(MPFC)中CRF在调节酗酒中的作用。 拟议的研究将采用生化、药物遗传学、转基因和行为药理学。 接近了。在啮齿动物“黑暗中饮酒”(DID)中，暴饮式酒精摄取的良好验证模型将是 在这些实验中使用。对于目标1，我将使用DID范例和免疫组织化学 蛋白质印迹技术评估1周或3周的DID是否改变了mPFC CRF系统的成分 CRF，CRF受体1和2(CRF1R/CRF2R)，CRF结合蛋白。对于目标2，我将使用Designer Receiver CRH-IRES-Cre转基因小鼠DID体内病毒注射CRE依赖的DREADDS技术 用于(1)确定是否通过激活抑制性DREADD使mPFC中的CRF神经元“沉默” 减少暴饮式酒精摄入量和相关的血液酒精浓度(BECs)和(2)评估 激活mPFC内CRF神经元(通过兴奋性DREADD)可减少暴饮式酒精摄入 和相关的BEC。对于目标3，我将使用行为药理学和特定部位的微量注射来评估 MPFC内CRF1R和CRF2R受体在调节狂欢样酒精中的相对作用 DID范例中的摄入量。这个项目的结果将促进我们对神经机制的理解。 潜在的暴饮式乙醇摄入，以及表征mPFC特定成分的潜力 CRF系统可作为酒精依赖治疗的靶点。

Project Summary/Abstract Alcohol dependence is a widespread public health concern with limited treatment options available. A growing body of evidence supports binge intake of ethanol as a significant risk factor for the development of ethanol dependence. Additionally, this form of excessive ethanol intake is associated with negative health effects, such as heart disease and diabetes, and represents a significant portion of the financial burden induced by alcohol use within the United States. Our lab and others have extensively demonstrated the neuropeptide corticotropin releasing factor (CRF) plays an important role in modulating binge ethanol consumption. Further, alterations in the CRF system are known to play an important role in ethanol-related behaviors both following the development of ethanol-dependence and into withdrawal. Dissecting the specific role of CRF in binge drinking behavior in non-ethanol dependent animals therefore enables the discovery of treatments for this risky pattern of behavior and, further, provides important insights into initial ethanol related alterations of a system critically involved in the development and maintenance of ethanol-dependence. The role of CRF in ethanol-related behaviors within the extended amygdala has been the focus of numerous investigations. However, the role of CRF within regions critical to entraining these limbic regions and coordinating reward-directed/inhibiting inappropriate behaviors has gone essentially neglected. This proposal focuses on addressing this hole in the literature by examining the role of CRF within the medial prefrontal cortex (mPFC) in modulating binge ethanol consumption. The proposed studies will employ biochemical, pharmacogenetic, transgenic, and behavioral pharmacological approaches. The well-validated model of binge-like ethanol intake in rodents ‘drinking in the dark’ (DID) will be used throughout these experiments. For Aim 1, I will use the DID paradigm and immunohistochemistry and western blot techniques to assess whether 1 or 3-weeks of DID alter components of the mPFC CRF system (CRF, CRF receptor 1 and 2 (CRF1R/CRF2R), CRF-binding protein). For Aim 2, I will use designer receptor technology (viral injection of Cre-dependent DREADDs) along with CRH-IRES-Cre transgenic mice in the DID model to (1) determine whether ‘silencing’ CRF neurons (via activation of inhibitory DREADDs) in the mPFC decreases binge-like ethanol intake and associated blood ethanol concentrations (BECs) and (2) assess whether activation of CRF neurons (via excitatory DREADDs) in the in the mPFC decreases binge-like ethanol intake and associated BECs. For Aim 3, I will use behavioral pharmacology and site-specific microinjections to assess the relative contribution of the CRF1R and CRF2R receptors within the mPFC in modulating binge-like ethanol intake in the DID paradigm. Results from this project will advance our understanding of the neural mechanisms underlying binge-like ethanol intake, as well as characterize the potential for specific components of the mPFC CRF system to serve as a therapeutic target for treatment of alcohol dependence.