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)在女性中的患病率是男性的两倍， 个人也会患上酒精使用障碍(AUD)。将捕食者气味应激(PS)用作动物 在创伤后应激障碍模型中，我们确定两次PS暴露显著增加焦虑相关行为和 雄性和雌性C57BL/6J(B6)小鼠海马神经元的激活。值得注意的是，间歇性PS 酒精(酒精)摄入量显著增加60%(男性)和71%(女性)，在 回应。此外，对PS增强的乙醇摄入量的“敏感性”显著增加促肾上腺皮质激素。 女性与男性HC中释放因子受体-1(CRFR1)的蛋白水平，与性别证据一致 逆境后CRFR1信号的差异。拟议的研究以上述证据为基础，通过测试 PTSD和AUD共病的假设是由于HC中CRFR1表达增加所致 投射到mPFC的神经元和PS诱导CRFR1的性别差异参与了这一过程 合并症。目标1将确定在PS增强的关联中是否存在性别差异 饮酒对B6小鼠焦虑、心率和/或强迫饮酒的影响。我们 预测“敏感”小鼠的PS增强饮酒将与焦虑、心率和 并认为强制饮酒会有性别差异的模式发生变化。AIM 2将绘制出 Crfr1-GFP小鼠的CRFR1表达和PS及PS增强饮酒对神经元的激活作用。我们预测 大脑局部CRFR1标记的活动模式会有性别差异 间歇性PS与酒精摄入量的关系。目标3将操纵CRFR1的活性- 用化学遗传学或药理学方法表达神经元，并确定对PS-1的影响 增加了饮酒量。两项研究将使用专门由设计师药物激活的设计师受体 (DREADDS)检测CRFR1在腹侧CA1区的必要性和充分性，并抑制(GI) 和兴奋性(GQ)DREADDS。我们预测，阻止PS诱导的腹侧CA1激活 (GI DREADD)将仅在“敏感”小鼠中阻断PS增强的饮酒，而激活腹侧CA1(GQ) DREADD)将增加饮用无间歇性PS的乙醇小鼠的乙醇摄入量。一项补充研究 将确定全身应用CRFR1拮抗剂是否会减少PS增强的饮酒 在B6小鼠中摄入，并预测该拮抗剂将在“敏感”的小鼠中最有效。目标4将 确定处理从腹侧CA1到mPFC的投射对于PS增强是否重要 在B6小鼠中，通过向腹侧CA1注射GI DREADDS和向mPFC注射氯氮平-N-氧化物。我们预测 阻止PS诱导的腹侧CA1至mPFC投射的激活将阻止PS增强的饮酒。 总而言之，这些信息将阐明PS敏感机制的性别差异。 增加饮酒可作为治疗创伤后应激障碍所致的AUD的靶点。

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.