Transgenerational Inheritance of a Cocaine Resistance Phenotype

项目摘要 这项自2012年以来一直由美国国立卫生研究院资助的研究项目的重点是父亲的影响 心理刺激性自我给药对后代(即后代)生理和行为的影响 和子孙)使用大鼠模型。我们之前的研究表明，父亲的可卡因自我给药 对生殖系表观基因组进行了重新编程，导致可卡因在雄性后代中的增强效果降低。 目前的应用扩大了我们对可卡因的关注范围，以涵盖可卡因的跨代影响 父亲的甲基苯丙胺(冰毒)，将与可卡因进行比较和对比。建议数 研究将检查信息传递给冰毒后代的机制(并可能 孙代)通过精子的表观遗传学变化。我们还将定义表观遗传和转录 可卡因和冰毒后代伏隔核中可能分别构成 对心理刺激性自我管理的影响。具体目标1将检查以下行为后果 父性冰毒自我给药对雌雄后代精神刺激性自我给药的影响(F1)和 孙代(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结果 心理刺激剂对雄性后代增强效能的影响。与我们之前的可卡因实验结果不同， 初步数据表明，父亲的自我管理导致自我管理的增加，和 动机，这种精神刺激剂在雄性后代中选择性地存在。药物幼稚的F1大鼠将被用来产生 F2代，将评估冰毒的获取、维持和增强效力，就像 在F1后代中。在特定目标2中，我们将评估精子的表观遗传学变化 心理刺激性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Will表观遗传学改变(小的非编码RNA和DNA甲基化) 被评估。最后，对心理刺激跨代效应的全基因组评估还没有 对伏隔核中的神经元进行了实验，伏核是大脑中在调制过程中起关键作用的区域 心理刺激剂诱导的行为。具体目标3中的实验旨在询问 单核ATAC-SEQ和单核偶联的可及染色质景观和基因表达 实验用纯冰毒、可卡因和生理盐水繁殖的大鼠伏隔核中的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 检查心理刺激相关表观遗传机制的行为方法学 信息可以从父代传递给后代和孙辈。这些跨代研究 代表了一种新的策略来识别与心理刺激自我保护因素相关的转录本 这将为治疗开发提供新的靶点。

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.