Sensitivity and resilience to increased alcohol drinking in males and females following traumatic stress

项目摘要 创伤后应激障碍（PTSD）在女性中的发病率是男性的两倍， 也发展为酒精使用障碍（AUD）的人。使用捕食者气味应激（PS）作为动物 在PTSD模型中，我们确定两次PS暴露显著增加了焦虑相关行为， 雄性和雌性C57 BL/6J（B6）小鼠海马（HC）中的神经元活化。值得注意的是，间歇性PS 酒精（乙醇）摄入量显著增加60%（男性）和71%（女性）， 反应此外，对PS增强的乙醇摄入的"敏感性"赋予显着更高的促肾上腺皮质激素 女性HC与男性HC的释放因子受体-1（CRFR1）蛋白水平，与性别证据一致 应激后CRFR1信号传导的差异。拟议的研究建立在上述证据的基础上， 假设PTSD和AUD的共病是由于HC中CRFR1表达增加 神经元投射到mPFC，PS诱导CRFR1的性别差异对此有贡献 科摩罗。目的1将确定是否存在性别差异的关联PS增强 B6小鼠中的乙醇饮用和焦虑、心率（HR）和/或强迫性乙醇饮用的改变。我们 预测PS增强饮酒在"敏感"小鼠将与焦虑，心率， 并认为强迫性饮酒的模式变化会有性别差异。Aim 2将映射 在crfr1-gfp小鼠中，PS和PS增强饮水引起的CRFR1表达和神经元激活。我们预测 大脑区域CRFR1-collabeled活动模式将存在性别差异， 间歇性PS与乙醇摄入量的关系。目的3将操纵CRFR1的活性- 表达神经元，并确定对PS的影响。 强化饮酒两项研究将使用专门由设计药物激活的设计受体 （DREADDs），以测试腹侧CA 1中CRFR1的必要性和充分性，抑制（Gi） 和兴奋性（Gq）DREADDs。我们预测，阻止PS诱导的腹侧CA1激活， (Gi DREADD）将阻止PS增强饮水只在“敏感”小鼠，而激活腹侧CA1（Gq DREADD）将增加饮用乙醇而无间歇性PS的小鼠的乙醇摄入。补充研究 将确定全身给予CRFR1拮抗剂是否会减少PS增强的饮酒 B6小鼠中的摄入量，预测拮抗剂在"敏感"小鼠中最有效。目标4将 确定腹侧CA1到mPFC的投射操作是否对PS增强的 B6小鼠通过腹侧CA1注射Gi DREADD和mPFC注射氯氮平-N-氧化物。我们预测 阻止PS诱导的腹侧CA1向mPFC投射的激活将阻止PS增强的饮酒。 总的来说，这些信息将阐明对PS敏感性机制的性别差异， 增强饮酒，可用于治疗PTSD诱导的AUD。

Project Summary Post-traumatic stress disorder (PTSD) is twice as prevalent in females as in males, with a proportion of individuals also developing an alcohol use disorder (AUD). Using predator odor stress (PS) as an animal model of PTSD, we determined that two PS exposures significantly increased anxiety-related behavior and neuronal activation in the hippocampus (HC) of male and female C57BL/6J (B6) mice. Notably, intermittent PS significantly increased alcohol (ethanol) intake by 60% (males) and 71% (females), with heterogeneity in the response. Further, “sensitivity” to PS-enhanced ethanol intake conferred significantly greater corticotropin releasing factor receptor-1 (CRFR1) protein levels in female versus male HC, consistent with evidence for sex differences in CRFR1 signaling following stress. The proposed studies build on the above evidence by testing the hypothesis that comorbidity of PTSD and AUD is due to increased CRFR1 expression in HC neurons projecting to mPFC and that sex differences in CRFR1 induction by PS contribute to this comorbidity. Aim 1 will determine whether sex differences exist in the association between PS-enhanced ethanol drinking and alteration in anxiety, heart rate (HR), and/or compulsive ethanol drinking in B6 mice. We predict that PS-enhanced drinking in “sensitive” mice will be associated with an increase in anxiety, HR, and compulsive drinking and that there will be sex differences in the pattern of changes. Aim 2 will map changes in CRFR1 expression and neuronal activation by PS and by PS-enhanced drinking in crfr1-gfp mice. We predict that there will be sex differences in brain regional CRFR1-colabelled activity patterns in response to intermittent PS and in the relationship with ethanol intake. Aim 3 will manipulate the activity of CRFR1- expressing neurons using chemogenetic or pharmacologic approaches and determine the impact on PS- enhanced drinking. Two studies will use Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) in crfr1-cre mice to test the necessity and sufficiency of CRFR1 in ventral CA1, with inhibitory (Gi) and excitatory (Gq) DREADDs, respectively. We predict that preventing PS-induced activation of ventral CA1 (Gi DREADD) will block PS-enhanced drinking only in “sensitive” mice, whereas activating the ventral CA1 (Gq DREADD) will enhance ethanol intake in mice drinking ethanol without intermittent PS. A complementary study will determine whether systemic administration of a CRFR1 antagonist will reduce PS-enhanced drinking intake in B6 mice, with the prediction that the antagonist will be most effective in “sensitive” mice. Aim 4 will determine whether manipulation of the projection from ventral CA1 to mPFC is important for PS-enhanced drinking in B6 mice, by injecting Gi DREADDs into ventral CA1 and clozapine-N-oxide into mPFC. We predict that preventing PS-induced activation of the ventral CA1 to mPFC projection will block PS-enhanced drinking. Collectively, the information will elucidate sex differences in mechanisms underlying sensitivity to PS- enhanced drinking that can be targeted for the treatment of PTSD-induced AUD.