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

基本信息

批准号：
8847311
负责人：
ELIZABETH ANNE EIPPER
金额：
$ 19.63万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8847311
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 Health 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&apos 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 response serotonin receptor sex therapeutic target therapy design tool transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): 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 effets 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.

描述（由申请人提供）：从最初的生理反应、寻求药物或自我给药行为的获得速度以及复发倾向的强度来看，女性和雌性实验动物对可卡因的敏感度远高于男性和雄性实验动物。众所周知，女性在许多情绪和认知反应上与男性不同，但我们对这些差异的分子基础的理解是有限的。我们将使用深度测序来深入了解导致女性和男性对可卡因戒断反应不同的机制。我们之前将这种方法应用于从实验者施用可卡因中撤出的雄性小鼠的伏核，并确定了 Wnt/钙粘蛋白途径和 miR-8 家族成员作为关键参与者。对特定通路的靶向分析揭示了可卡因介导的伏隔核中编码多巴胺、谷氨酸、GABA、乙酰胆碱、神经肽和内源性大麻素信号通路多种成分的 mRNA 表达变化。在目标 1 中，将检查四组小鼠：雄性、循环雌性、切除卵巢的雌性和用雌二醇替代切除卵巢的雌性。小鼠注射生理盐水或可卡因一周，停药四周后处死；将收获伏隔核、前额皮质和腹侧被盖区用于制备RNA。从表现出运动敏化的小鼠伏核中制备的重复条形码文库将通过技术复制同时进行测序。生物信息学分析将用于识别所有群体共有的可卡因反应转录本和途径（核心可卡因反应）、女性独有且对雌激素敏感。在目标 2 中，我们将从核心可卡因反应组和女性敏感性组中选择转录本和通路进行验证和进一步分析。该数据集将允许分析性、雌激素和可卡因对选择性剪接和 RNA 编辑的影响。将对可卡因反应基因启动子区域的雌激素反应元件和转录因子结合位点进行生物信息学分析。有了这个高质量、经过验证的数据集，重点测序研究可用于分析个体小鼠自我施用可卡因的反应。这种广泛的方法应该能够识别女性特有的或对雌激素敏感的治疗靶点。