Exploiting mouse models to understand female hypersensitivity to cocaine

利用小鼠模型了解女性对可卡因的过敏反应

• 批准号: 8630003
• 负责人: ELIZABETH ANNE EIPPER
• 金额: $ 23.85万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8630003
• 关键词: Acetylcholine; Alternative Splicing; Area; Bar Codes; Behavior; Behavioral; Binding Sites; Bioinformatics; Brain region; Cadherins; Chronic; Cocaine; Cognitive; Data; Data Set; Dopamine; Elements; Emotional; Endocannabinoids; Estradiol; Estrogen Replacements; Estrogens; Exhibits; Family member; Female; G-Protein Signaling Pathway; Gene Expression Profile; Genes; Glutamates; Harvest; Heterotrimeric GTP-Binding Proteins; Hypersensitivity; Individual; Laboratory Animals; Libraries; Mediating; Methyl-CpG-Binding Protein 2; MicroRNAs; Molecular; Mus; Neuropeptides; Nucleus Accumbens; Pathway Analysis; Pathway interactions; Pharmaceutical Preparations; Physiological; Play; Prefrontal Cortex; Preparation; Promoter Regions; RNA; RNA Editing; RNA Splicing; Reading; Relapse; Research Personnel; Response Elements; Rodent; Role; Saline; Self Administration; Self-Administered; Sequence Analysis; Signal Pathway; Signal Transduction; Transcript; Validation; Ventral Tegmental Area; Western Blotting; Withdrawal; Woman; Women' s Group; base; cholinergic; chromatin modification; deep sequencing; drug of abuse; drug seeking behavior; gamma-Aminobutyric Acid; insight; interest; mRNA Expression; male; men; mouse model; neurochemistry; protein expression; public health relevance; response; serotonin receptor; sex; therapeutic target; therapy design; tool; transcription factor; transcriptome sequencing

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.

从最初的生理反应判断，药物寻求或自我寻求的获得速度 给药行为和复发倾向的强度，女性和雌性实验室动物是 对可卡因的敏感性远高于人类和雄性实验动物。虽然众所周知， 在许多情感和认知反应上与男性不同，我们对分子的理解 这些差异的基础有限。我们将使用深度测序来深入了解 导致在女性中观察到的可卡因戒断反应不同的机制， 男性。我们先前将这种方法应用于雄性小鼠的脑桥核， 实验者给予可卡因并鉴定了Wnt/钙粘蛋白通路和miR-8家族成员 作为关键球员。对特定通路的靶向分析揭示了可卡因介导的 表达编码多巴胺、谷氨酸、GABA、乙酰胆碱 神经肽和内源性大麻素信号通路的研究。 在目标1中，将检查四组小鼠：雄性、周期性雌性、卵巢切除雌性 雌二醇替代了切除卵巢的雌性。注射生理盐水或可卡因一周的老鼠将被 停药4周后处死;丘脑核、前额叶皮质和腹侧被盖区 将收获用于制备RNA。从细胞核制备的重复条形码文库 将同时对表现出运动致敏的小鼠的染色体进行测序， 复制品。生物信息学分析将用于识别可卡因反应转录本和途径 所有群体都有（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途径。 敏感性组进行验证和进一步分析。数据集将允许分析性别的影响， 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 可卡因应答基因启动子区的应答元件和转录因子结合位点 基因将被执行。 有了这个高质量的、经过验证的数据集，可以使用集中的测序研究来分析基因组。 个体小鼠自我施用可卡因的反应。这一广泛的方法应允许确定 女性特有的或对雌激素敏感的治疗靶点。