Stress, alcohol and GABAergic neuroactive steroids in primates

描述（由申请人提供）：本提案的目的是探索灵长类动物血浆中应激反应、饮酒和神经活性类固醇水平之间的关系。啮齿动物模型中的大量证据表明，GABA能神经活性类固醇有助于乙醇敏感性、耐受性、防止依赖性并减少过量饮酒。灵长类动物合成不同的类固醇前体并表现出5 α-前体酶活性，这需要开发高度灵敏的GCMS测定来测量灵长类动物血浆中孕酮、脱氧皮质酮和皮质醇的3 α 5 β和3 α 5 β-还原代谢物。待检验的总体假设是，在未接触过乙醇的猴中激活HPA轴和/或给予乙醇后，灵长类动物血浆中的GABA能神经活性类固醇升高，但在慢性乙醇暴露后丢失。将研究下丘脑垂体肾上腺轴的药理学激活、酒精输注、饮酒诱导和慢性饮酒的影响，以确定神经活性类固醇对HPA轴激发的反应与饮酒倾向之间是否存在相关性。第一个目标是开发一种高灵敏度的GCMS分析方法，用于测量灵长类动物血浆中孕酮、脱氧皮质酮和皮质醇的3a 5 α-和3a 5 [3]-还原代谢物。第二个目的是确定在下丘脑（纳洛酮给药）、垂体（oCRF输注）或肾上腺（地塞米松预处理后ACTH输注）水平的HPA轴刺激和全身乙醇给药是否改变猴血浆中GABA受体神经活性类固醇的水平。第三个目标是研究使用相同的激发程序刺激HPA轴后，计划诱导的饮酒和随意饮酒一年对前体类固醇和神经活性类固醇水平的影响。第四个目标将为其他INIA研究人员提供服务核心，探索长期乙醇暴露对小鼠，猕猴和酗酒者神经类固醇水平的影响。每名研究者将探索对乙醇或HPA轴刺激的神经活性类固醇反应是否因慢性乙醇史或预测自愿饮酒水平而改变。

DESCRIPTION (provided by applicant): The purpose of this proposal is to explore the relationship between stress response, alcohol drinking and neuroactive steroid levels in primate plasma. A large body of evidence in rodent models suggests that GABAergic neuroactive steroids contribute to ethanol sensitivity, tolerance, protection against dependence and reduces excessive alcohol consumption. Primates synthesize different steroid precursors and exhibit 5preductase activity that necessitates the development of a highly sensitive GCMS assay to measure both 3<x5ccand 3a5p- reduced metabolites of progesterone, deoxycorticosterone and cortisol in primate plasma. The overall hypothesis to be tested is that GABAergic neuroactive steroids are elevated in primate plasma following activation of the HPA axis and/or ethanol administration in ethanol nai've monkeys, but lost after chronic ethanol exposure. The effects of pharmacological activation of the hypothalamic pituitary adrenal axis, alcohol infusion, induction of alcohol drinking and chronic alcohol consumption will be investigated to determine if there is an association between neuroactive steroid responses to HPA axis challenge and the propensity to drink alcohol. These studies will be integrated into the INIA consortium by comparison with similar studies in mice and man. The first aim will develop a highly sensitive GCMS assay to measure both 3a5a- and 3a5[3- reduced metabolites of progesterone, deoxycorticosterone and cortisol in primate plasma. The second aim will determine if HPA axis stimulation at the level of the hypothalamus (naloxone administration), pituitary (oCRF infusion) or adrenal gland (ACTH infusion after dexamethasone pretreatment) and systemic ethanol administration alters the levels of GAB A receptor neuroactive steroids in plasma of monkeys. The third aim will investigate the effect of schedule-induced drinking and ad lib ethanol consumption for one year on both the precursor steroids and neuroactive steroid levels following HPA axis stimulation using the same challenge procedure. The fourth aim will provide a service core to other INIA investigators exploring the effects of chronic ethanol exposure on neurosteroid levels in mice, macaque monkeys and alcoholic humans. Each investigator will explore if neuroactive steroid responses to ethanol or HPA axis stimulation are altered by chronic ethanol history or predictive of voluntary alcohol consumption levels.