Optogenetic studies of hypocretin in binge drinking and negative hedonic valence

描述(申请人提供)：酒精中毒是一种慢性疾病，具有严重的社会后果，而压力在导致酒精依赖者酒精消费复发(Etoh)方面起着关键作用。即使在非依赖型人群中，也有证据支持神经应激网络在酒精过量摄入中起到关键作用。因此，需要进一步的研究来推进关于压力相关行为和过量饮酒背后复杂的神经生物学的科学知识。事实上，多个应激神经肽系统以复杂的方式促进了乙醇相关的行为。C57BL/6J(B6)小鼠是研究过量乙醇摄入的神经生物学的理想模型，因为它们在昼夜暗周期的离散时期内自愿摄入足够数量的乙醇，从而产生超过NIAAA的狂饮标准(80 mg/dL或0.08%)的血液乙醇浓度(BECs)。这项建议将把体内的光遗传刺激/抑制实验与长期间歇性狂饮和厌恶场所条件作用的模型结合起来，以建立(或驳斥)下丘脑下丘脑下克隆素系统、应激性高觉醒状态和病理性乙醇消耗之间的因果关系。此外，这些实验将使用病毒示踪和双重荧光免疫组织化学从解剖学上定义连接促肾上腺皮质激素释放因子(CRF)和下丘脑泌素系统的通路，并确定这些相互作用如何控制应激反应和乙醇饮酒行为。应激激素测量、亚型特异性药理学和转录因子定位将提供补充措施来证实这些发现，并进一步确定这些行为背后的精确神经底物。从拟议的实验中获得的知识将为未来减少药物滥用和复发的战略提供依据。

DESCRIPTION (provided by applicant): Alcoholism is a chronic disease with severe consequences to society, and stress plays a key role in precipitating relapse of ethanol (EtOH) consumption in dependent alcoholics. Even in non-dependent populations, evidence supports a key role for neural stress networks in binge EtOH intake. Therefore, further research is required to advance scientific knowledge of the intricate neurobiology underlying stress-related behavior and excessive EtOH drinking. Indeed, multiple stress neuropeptide systems contribute to EtOH- related behaviors in complex ways. C57BL/6J (B6) mice serve as an ideal for model interrogating the neurobiology of excessive EtOH intake, as they voluntarily consume sufficient quantities of EtOH within discrete periods of the circadian dark cycle to produce blood EtOH concentrations (BECs) that surpass the NIAAA's criteria for binge drinking (80 mg/dL, or .08 percent). This proposal will integrate in vivo optogenetic stimulation/inhibition experiments with models of long-term intermittent binge drinking and aversive place conditioning in order to establish (or refute) causal relationships between the hypothalamic hypocretin system, stress-like states of hyperarousal, and pathological EtOH consumption. In addition, these experiments will use viral tracing and double fluorescent immunohistochemistry to anatomically define the pathways connecting the corticotropin-releasing factor (CRF) and hypocretin systems, and determine how these interactions control the stress response and EtOH drinking behavior. Stress hormone measurements, subtype-specific pharmacology, and transcription factor mapping will provide complementary measures to confirm the findings and further define the precise neural substrates underlying these behaviors. Knowledge gained from the proposed experiments will inform future strategies for mitigation of drug abuse and relapse.