Gene Delivery of Neuroactive Steroids to Modulate Ethanol Reinforcement/Consumption

项目摘要 动物和人类研究表明，神经活性类固醇的升高可能会解决许多行为问题， 与酒精使用障碍相关的病理学。这个项目的目标是评估假设， 腹侧被盖区类固醇生成增加将减少操作性乙醇自我给药 以及男性和女性酒精偏好者在被剥夺后自愿饮酒的增加（P） 大鼠内源性神经活性类固醇将通过病毒载体介导的基因递送而升高， 生物合成酶P450 SCC，将胆固醇转化为双烯醇酮。我们最近的研究表明， 载体介导的P450 SCC向腹侧被盖区的传递减少了乙醇的自我给药，并增加了局部的 （3α，5 α）-3-羟基异孕烷-20-酮（3α，5 α-THP，别孕烯醇酮）的表达（Cook等人，2014）。我们现在 我建议通过检查雄性和雌性P大鼠的影响来扩展这些研究，探索对 剥夺诱导的饮酒并将载体靶向VTA中的酪氨酸羟化酶（TH）神经元。我们将 检查媒介和行为的特异性以及影响的持久性。目标1将调查，如果 通过将P450 SCC基因递送至VTA来提高类固醇生成改变A）操作性乙醇自我给药 在非依赖性雄性和雌性P大鼠中，或B）乙醇依赖性雄性中的剥夺诱导饮酒， 雌性P大鼠。目的2将检测腹侧被盖区TH神经元特异性类固醇生成的升高是否改变A） 非依赖性雄性和雌性大鼠的操作性乙醇自我给药或B）剥夺诱导的饮酒 在酒精依赖的雄性和雌性P大鼠中。这些研究将增加我们对VTA作用的理解 神经活性类固醇在乙醇强化、焦虑样行为和乙醇后饮酒量增加中的作用 剥夺

Project Summary Animal and human studies suggest that elevation of neuroactive steroids may address many of the behavioral pathologies associated with alcohol use disorders. The goal of this project is to evaluate the hypotheses that elevated steroidogenesis in the ventral tegmental area will reduce operant ethanol self-administration and the escalation of voluntary drinking following deprivation in male and female alcohol preferring (P) rats. Endogenous neuroactive steroids will be elevated by viral vector-mediated gene delivery of the biosynthetic enzyme P450scc that converts cholesterol to pregnenolone. Our recent studies demonstrate that vector-mediated delivery of P450scc to the VTA reduces ethanol self-administration and increases local expression of (3α,5α)-3-hydroxypregnan-20-one (3α,5α-THP, allopregnanolone) (Cook et al., 2014). We now propose to extend these studies by examining effects in both male and female P rats, probing effects on deprivation-induced drinking and targeting the vector to tyrosine hydroxylase (TH) neurons in the VTA. We will examine vector and behavioral specificity as well as the persistence of effects. Aim 1 will investigate if elevation of steroidogenesis by gene delivery of P450scc to VTA alters A) operant ethanol self-administration in non–dependent male and female P rats or B) deprivation-induced drinking in ethanol dependent male and female P rats. Aim 2 will examine whether TH neuron-specific elevation of steroidogenesis in VTA alters A) operant ethanol self-administration in non–dependent male and female rats or B) deprivation-induced drinking in ethanol dependent male and female P rats. These studies will increase our understanding of the role of VTA neuroactive steroids in ethanol reinforcement, anxiety-like behavior and escalated drinking following ethanol deprivation.