Cocaine Regulation of miRNAs in Rats with Differing Vulnerability to Drug Abuse

可卡因对不同药物滥用易感性大鼠中 miRNA 的调节

• 批准号: 7586020
• 负责人: ROBERT C THOMPSON
• 金额: $ 36.9万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7586020
• 关键词: Abstinence; Acute; Address; Adult; Affect; Animals; Attention; Behavior; Behavioral; Biochemistry; Brain; Brain region; Brain-Derived Neurotrophic Factor; Breeding; Cells; Cocaine; Cocaine Dependence; Condition; Coping Behavior; Data; Development; Dimensions; Dose; Drug Exposure; Drug abuse; Drug usage; Endowment; Ensure; Exhibits; Exposure to; Functional RNA; Gene Expression; Gene Targeting; Genes; Genetic; Growth Factor; Hippocampus (Brain); Human; In Situ Hybridization; Individual Differences; Lasers; Lead; Light; Link; Maintenance; Medial; Mediating; Mediator of activation protein; Messenger RNA; Methods; MicroRNAs; Microscopy; Molecular; Monitor; Morphology; Neurobiology; Neuronal Plasticity; Neurons; Nucleus Accumbens; Pattern; Pharmaceutical Preparations; Phenotype; Physiology; Play; Predisposing Factor; Prefrontal Cortex; Procedures; Process; Production; Proteins; Protocols documentation; Public Health; Rattus; Regulation; Research; Rewards; Rodent; Role; Sampling; Self Administration; Self-Administered; Signal Transduction; Staging; Stress and Coping; Substance abuse problem; Synapses; Synaptic plasticity; Testing; Thinking; Time; Tissues; Transcript; Translations; Ventral Tegmental Area; Vertebral column; Work; base; brain behavior; drug induced behavior; drug of abuse; genetic element; neural circuit; neurogenesis; novel; protein expression; psychologic; response; reward processing; stimulant abuse; synaptogenesis; trait

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs), small (~22nt non-coding RNAs) function as sequence-specific post-transcriptional regulators of gene expression, by reducing target mRNA levels and/or inhibiting protein translation. Schratt et.al. (2006) have shown a link between miRNA function, neuronal spine development and the action of at least one growth factor that may play a role in neuroplasticity. Yet, little is known about miRNA expression patterns in the adult brain, or whether changes in miRNA expression occur following either acute or repeated drug use or whether any miRNA might underlie any known messenger RNA (or protein) changes known to occur following drugs use (or abuse). We hypothesize that miRNA expression alterations will be distinct across various stages of substance abuse and that drug exposure will result in distinctive miRNA expression patterns (anatomical and/or temporal) in two distinct groups of rodents selectively-bred based upon the novelty seeking trait (High Responders (HR) vs Low Responders (LR)), as these two groups of rodents have unique patterns of gene expression basally, after various types of challenges, and display individual differences in the propensity for substance abuse. In Aim 1, we will use laser capture microscopy to isolate specific brain regions known to be direct targets of cocaine and/or undergo drug-induced changes in synaptic plasticity prior to broad-scale miRNA profiling. We will use tissues derived from both HR and LR rodents under basal conditions; conditions where gene expression differences exist. miRNA expression differences will be validated by miRNA qPCR and spatial patterns of expression analyzed by miRNA in situ hybridization (ISH) methods. In aim two, we will use a cocaine sensitization paradigm (where drug exposure can be held constant) in both HR and LR rodents to examine miRNA regulation by cocaine at two time points (immediately following cocaine and following a period of abstinence) isolated and monitored as in aim 1. In aim 3, we will use cocaine self-administration procedures in HR and LR rodents, collect tissues at two time points (acute and abstinence) to examine miRNA regulation. In Aim 4, we will use antagomirs to block basal or cocaine regulated HR/LR miRNA differences to determine if miRNA blockade alters drug-induced behaviors. Together, these findings will shed light on whether miRNAs contribute to the initial propensity to seek drugs of abuse or play an important role in the induction and/or maintenance of cocaine addiction. PUBLIC HEALTH RELEVANCE: Our project evaluates small but novel genetic elements called microRNAs (miRNAs) that are thought to have broad influences on the types or amounts of proteins cells produce. In this proposal, we plan to examine whether there are miRNA changes in two different rodent lines that vary in their propensity to seek drugs under basal conditions. Further, we plan to examine in each rodent line how different miRNA levels vary in multiple brain regions as a function of cocaine use both acutely as well as long after the drug treatment has ceased. These findings will shed light on whether miRNAs contribute to the initial propensity to seek drugs of abuse or play an important role in the induction and/or maintenance of cocaine addiction.

描述(由申请人提供)：microRNAs(MiRNAs)，小的(~22nt非编码RNAs)作为序列特异性的转录后调节基因表达，通过降低目标mRNA水平和/或抑制蛋白质翻译。Schratt等人。(2006)的研究表明，miRNA功能、神经元脊椎发育和至少一种可能在神经可塑性中发挥作用的生长因子的作用之间存在联系。然而，人们对成人大脑中miRNA的表达模式知之甚少，也不知道miRNA表达的变化是否发生在急性或重复吸毒后，或者是否有miRNA可能是已知的吸毒(或滥用)后发生的信使RNA(或蛋白质)变化的基础。我们假设，miRNA的表达变化在药物滥用的不同阶段将是不同的，并且药物暴露将导致两组不同的啮齿动物(解剖和/或时间)的miRNA表达模式不同，这两组啮齿动物是基于寻求新颖性的特征(高响应者(HR)和低响应者(LR))培育的，因为这两组啮齿动物在经历了各种类型的挑战后，基本上具有独特的基因表达模式，并在药物滥用倾向方面显示出个体差异。在目标1中，我们将使用激光捕获显微镜来分离已知为可卡因直接目标的特定大脑区域和/或在大规模miRNA分析之前经历药物诱导的突触可塑性变化。我们将在基础条件下使用来自HR和LR啮齿动物的组织，即存在基因表达差异的条件。MiRNA表达差异将通过miRNA qPCR验证，并通过miRNA原位杂交(ISH)方法分析表达的空间模式。在目标2中，我们将在HR和LR啮齿动物中使用可卡因敏化范式(药物暴露可以保持恒定)来检测在目标1中分离和监测的两个时间点(紧随可卡因之后和戒断之后)可卡因对miRNA的调节。在目标3中，我们将在HR和LR啮齿动物中使用可卡因自我给药程序，在两个时间点(急性和戒断)收集组织来检测miRNA调节。在目标4中，我们将使用抗配子来阻断基础或可卡因调节的HR/LR miRNA差异，以确定miRNA阻断是否改变药物诱导的行为。总之，这些发现将阐明miRNAs是否有助于最初寻求滥用药物的倾向，或者在诱导和/或维持可卡因成瘾方面发挥重要作用。 与公共健康相关：我们的项目评估了被称为microRNAs(MiRNAs)的微小但新颖的遗传元素，这些基因元素被认为对细胞产生的蛋白质的类型或数量具有广泛影响。在这项建议中，我们计划检查两个不同的啮齿动物品系是否存在miRNA的变化，这两个品系在基础条件下寻找药物的倾向不同。此外，我们计划研究在每个啮齿类动物中，不同的miRNA水平如何随着可卡因的急性使用和药物治疗停止后的长期使用而在多个大脑区域发生变化。这些发现将阐明miRNAs是否有助于最初寻求滥用药物的倾向，或者在诱导和/或维持可卡因成瘾方面发挥重要作用。