Exploiting mouse models to understand female hypersensitivity to cocaine

从最初的生理反应判断，获得毒品或自我的速度 管理行为和复发倾向的强弱，女性和雌性实验动物 比男性和雄性实验动物对可卡因敏感得多。虽然众所周知，女性 与男性在许多情感和认知反应上的不同，我们对分子的理解 这些差异的基础是有限的。我们将使用深度排序来深入了解 女性和女性对可卡因戒断反应不同的机制 男性。我们之前将这种方法应用于雄性小鼠的伏隔核，这些小鼠从 实验人员注射了可卡因，并确定了Wnt/钙粘素途径和miR-8家族成员 作为关键的参与者。对特定通路的靶向分析揭示了可卡因介导的 编码多巴胺、谷氨酸、GABA、乙酰胆碱、 伏核中的神经肽和内源性大麻素信号通路。 在目标1中，将检查四组小鼠：雄性、骑自行车的雌性和去卵巢的雌性 雌二醇取代了去卵巢的女性。注射生理盐水或可卡因一周的小鼠将被 停药四周后处死动物：伏隔核、前额叶皮质和腹侧被盖区 将被收获用于制备RNA。从细胞核中制备的复制条形码文库 表现出运动敏感症的小鼠的伏隔神经将同时测序，技术上 复制。生物信息学分析将用于识别可卡因反应转录本和通路 所有群体都有共性(核心可卡因反应)，女性特有，对雌激素敏感。 在目标2中，我们将从核心可卡因反应组和女性中选择转录本和通路 敏感组进行验证和进一步分析。该数据集将允许分析性的影响， 雌激素和可卡因对选择性剪接和RNA编辑的影响。雌激素的生物信息学分析 可卡因反应性启动子区域的反应元件和转录因子结合位点 基因将被承担。 有了这个高质量的、经过验证的数据集，重点测序研究可以用来分析 自身注射可卡因的小鼠个体的反应。这一广泛的方法应该允许识别 女性特有或对雌激素敏感的治疗靶点。

Judging by initial physiological responses, rapidity of acquisition of drug-seeking or self- administration behavior, and strength of tendency to relapse, women and female laboratory animals are far more sensitive to cocaine than men and male laboratory animals. While it is well known that women differ from men in many emotional and cognitive responses, our understanding of the molecular underpinnings of these differences is limited. We will use deep sequencing to provide insight into the mechanisms that contribute to the different responses to withdrawal from cocaine observed in females and males. We previously applied this approach to the nucleus accumbens of male mice withdrawing from experimenter administered cocaine and identified the Wnt/cadherin pathway and miR-8 family members as key players. Targeted analysis of specific pathways revealed cocaine-mediated changes in the expression of mRNAs encoding multiple components of the dopamine, glutamate, GABA, acetylcholine, neuropeptide and endocannabinoid signaling pathways in the nucleus accumbens. In Aim 1, four groups of mice will be examined: males, cycling females, ovariectomized females and estradiol replaced ovariectomized females. Mice injected with saline or cocaine for a week will be sacrificed after four weeks of withdrawal; nucleus accumbens, prefrontal cortex and ventral tegmental area will be harvested for preparation of RNA. Duplicate bar-coded libraries prepared from the nucleus accumbens of mice exhibiting locomotor sensitization will be sequenced simultaneously, with technical replicates. Bioinformatic analysis will be used to identify cocaine-responsive transcripts and pathways common to all groups (core cocaine response), unique to females and sensitive to estrogen. In Aim 2 we will select transcripts and pathways from the core cocaine response group and female sensitivity group for validation and further analysis. The data set will allow analysis of the effects of sex, estrogen and cocaine on alternative splicing and RNA editing. Bioinformatic analysis of estrogen responsive elements and transcription factor binding sites in the promoter regions of cocaine-responsive genes will be undertaken. With this high quality, validated data set, focused sequencing studies can be used to analyze the response of individual mice self-administering cocaine. This broad approach should allow identification of therapeutic targets unique to females or sensitive to estrogen.