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: 8101347
• 负责人: ROBERT C THOMPSON
• 金额: $ 40.92万
• 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/8101347
• 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; 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; Rattus; Regulation; Regulator Genes; Research; Rewards; Rodent; Role; Sampling; Self Administration; Self-Administered; Signal Transduction; Staging; Stress and Coping; Substance abuse problem; Synapses; Synaptic plasticity; Testing; Time; Tissues; Transcript; Translations; Ventral Tegmental Area; Vertebral column; Work; base; brain behavior; cocaine exposure; cocaine use; drug induced behavior; drug of abuse; genetic element; neural circuit; neurogenesis; novel; protein expression; psychologic; public health relevance; 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）：微小RNA（miRNA）（约22nt非编码RNA）通过降低靶标mRNA水平和/或抑制蛋白质翻译，充当基因表达的序列特异性转录后调节剂。施拉特等人。 (2006) 已经表明 miRNA 功能、神经元脊柱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、急性或重复吸毒后是否发生 miRNA 表达的变化，或者是否有任何 miRNA 可能是吸毒（或滥用）后发生的任何已知信使 RNA（或蛋白质）变化的基础知之甚少。我们假设，在药物滥用的各个阶段，miRNA 表达的改变将是不同的，并且药物暴露将导致基于寻求新奇特征（高响应者（HR）与低响应者（LR））选择性培育的两组不同的啮齿动物中独特的 miRNA 表达模式（解剖学和/或时间性），因为这两组啮齿动物在经历各种类型的挑战后基本上具有独特的基因表达模式，并表现出个体差异 滥用药物的倾向。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；存在基因表达差异的条件。 miRNA 表达差异将通过 miRNA qPCR 进行验证，并通过 miRNA 原位杂交 (ISH) 方法分析表达的空间模式。在目标二中，我们将在 HR 和 LR 啮齿动物中使用可卡因致敏范式（其中药物暴露可以保持恒定），以检查可卡因在两个时间点（可卡因后立即和戒断一段时间后）对 miRNA 的调节，如目标 1 中那样分离和监测。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（急性期）收集组织。 和禁欲）来检查 miRNA 调控。在目标 4 中，我们将使用 antagomir 阻断基础或可卡因调节的 HR/LR miRNA 差异，以确定 miRNA 阻断是否会改变药物诱导的行为。总之，这些发现将揭示 miRNA 是否有助于最初寻求滥用药物的倾向，或者在诱导和/或维持可卡因成瘾中发挥重要作用。 公共健康相关性：我们的项目评估了称为 microRNA (miRNA) 的小型但新颖的遗传元件，这些元件被认为对细胞产生的蛋白质的类型或数量具有广泛的影响。在本提案中，我们计划检查两种不同的啮齿动物系中是否存在 miRNA 变化，这两种啮齿动物系在基础条件下寻求药物的倾向有所不同。此外，我们计划在每个啮齿动物系中检查不同 miRNA 水平在多个大脑区域中如​​何随可卡因急性使用以及药物治疗停止后很长时间内的变化而变化。这些发现将揭示 miRNA 是否有助于最初寻求滥用药物的倾向，或者在诱导和/或维持可卡因成瘾中发挥重要作用。