Transgenerational Inheritance of a Cocaine Resistance Phenotype

项目概要 该研究项目自 2012 年起一直由 NIH 资助，其重点是父亲的影响 精神兴奋剂的自我管理对后代（即后代）的生理和行为的影响 和孙辈）使用大鼠模型。我们之前的工作表明，可卡因的自我管理 重新编程了种系表观基因组，导致雄性后代的可卡因增强功效降低。 目前的申请扩大了我们对可卡因的关注范围，涵盖了可卡因的跨代影响 父亲甲基苯丙胺 (meth)，将与可卡因进行比较和对比。拟议的 研究将检查信息传递给吸食冰毒的后代的机制（并可能 孙子）通过精子的表观遗传变化。我们还将定义表观遗传和转录 可卡因和甲基苯丙胺后代的伏核轮廓可能是各自的基础 对精神兴奋剂自我给药的影响。具体目标 1 将检查以下行为的后果 父亲自我服用冰毒对男性和女性后代自我服用精神兴奋剂的影响（F1）和 孙辈（F2）。有趣的是，我们的初步结果表明冰毒和可卡因产生相反的作用 对男性后代精神兴奋剂强化功效的影响。与我们之前对可卡因的结果相比， 初步数据表明，父亲自我服用冰毒会导致自我服用冰毒的增加，并且 动机，这种精神兴奋剂选择性地作用于男性后代。未接受过药物治疗的 F1 级大鼠将用于产生 F2 代，其中冰毒的获取、维持和增强功效将被评估为 在F1后代中。在具体目标 2 中，我们将评估精子的表观遗传变化，通过这些变化父系 精神兴奋剂的自我管理可能会影响后代的行为。潜在的跨代 可卡因和冰毒诱导的精子表观遗传改变（小非编码 RNA 和 DNA 甲基化）将 进行评估。最后，精神兴奋剂跨代效应的全基因组评估尚未完成 对伏隔核中的神经元进行了研究，伏隔核是一个在调节中起着关键作用的大脑区域 精神兴奋剂引起的行为。具体目标 3 中的实验旨在询问 通过耦合单核 ATAC-seq 和单核获得可及染色质和基因表达的景观 对首次实验的甲基大鼠、可卡因大鼠和盐水大鼠的伏隔进行 RNA-seq 分析。 总的来说，本申请中描述的实验将使用最先进的细胞、分子和 行为方法学来检查表观遗传机制，从而精神兴奋剂相关 信息可以从父亲传递给后代和孙辈。这些跨代研究 代表了一种新的策略来识别与精神刺激自我的风险或保护因素相关的转录本 管理，这将阐明治疗开发的新目标。

Project Summary The focus of this research program, which has been NIH-funded since 2012, is the influence of paternal psychostimulant self-administration on the physiology and behavior of subsequent generations (i.e. offspring and grand-offspring) using rat models. Our work previously demonstrated that sire cocaine self-administration reprogramed the germline epigenome resulting in decreased cocaine reinforcing efficacy in the male progeny. The current application expands our focus on cocaine alone to encompass the transgenerational effects of paternal methamphetamine (meth), which will be compared and contrasted with cocaine. The proposed research will examine the mechanisms whereby information is passed to meth-sired offspring (and potentially grand-offspring) through epigenetic changes in sperm. We also will define epigenetic and transcriptional profiles in the nucleus accumbens of cocaine- and meth-sired offspring that may underlie the respective influences on psychostimulant self-administration. Specific Aim 1 will examine the behavioral consequences of paternal meth self-administration on psychostimulant self-administration in male and female offspring (F1) and grand-offspring (F2). Intriguingly, our preliminary results indicate that meth and cocaine produce opposite effects on psychostimulant reinforcing efficacy in male offspring. In contrast to our prior results with cocaine, preliminary data indicate that paternal meth self-administration results in increased self-administration of, and motivation for, this psychostimulant selectively in male offspring. Drug naïve F1 rats will be used to generate an F2 generation, where the acquisition, maintenance and reinforcing efficacy of meth will be assessed just as in F1 offspring. In Specific Aim 2 we will assess epigenetic changes in sperm through which paternal psychostimulant self-administration may influence the behavior of offspring. Potential transgenerational cocaine- and meth-induced epigenetic alterations (small noncoding RNAs and DNA methylation) in sperm will be evaluated. Finally, genome-wide assessments of psychostimulant transgenerational effects have not yet been performed on neurons in the nucleus accumbens, a brain region that plays a critical role in modulating psychostimulant-induced behaviors. The experiments in Specific Aim 3 are designed to interrogate the landscape of accessible chromatin and gene expression by coupling single-nuclei ATAC-seq and single-nuclei RNA-seq analyses in the accumbens of experimentally naive meth-sired, cocaine-sired and saline-sired rats. Collectively, the experiments described in this application will use state-of-the-art cellular, molecular and behavioral methodologies to examine epigenetic mechanisms whereby psychostimulant-associated information can be transmitted from sires to offspring and grand-offspring. These cross-generational studies represent a novel strategy to identify transcripts related to risk or protective factors for psychostimulant self- administration, which will illuminate new targets for therapeutic development.