Understanding cellular and molecular legacies of paternal stress

项目摘要：饥荒、大屠杀和其他自然和人为事件正在提供 有证据表明，创伤和压力的影响超越了祖辈，并影响了心理健康 在后代身上。纠正怀孕期间受到压力和缓解压力困扰的父母行为 因为它们在阻止这种压力遗产方面的效用而受到关注。相比之下，人们对如何 停止在受孕之前发生的父性压力的遗留。填补这一空白 知识，我们必须首先了解压力诱导的父系精子改变是如何扰乱神经生物学的 并破坏心理健康。有了这个意图，我们的目标是确定特定细胞类型的后代神经生物学 受到压力诱导的精子RNA变化的影响，这种变化已经成为一种机制，通过 父系血统给后代留下了压力的遗产。为了实现这一目标，我们依靠自己的经验 研究小鼠的父性压力、学习和记忆的遗传并建立在未发表的数据基础上 证明将应激暴露的雄性小鼠精子中的RNA注射到单细胞受精卵中 导致成年后在灭绝学习方面的缺陷。开始我们对神经生物学的研究 精子中的RNA启动这些灭绝学习缺陷的机制 暴露于压力之下，我们建议将重点放在下缘前额叶皮质的糖皮质激素受体(GRs)上 (IL-PFC)、IL-PFC内神经元基于乳酸的活性，以及IL-PFC的发展。我们的重点是塑造 通过以下背景。首先，IL-PFC对灭绝学习很重要。第二，表观遗传学 在研究代际关系的研究中，gr基因的调节受到了最多的关注。 在人类和啮齿动物中，由虐待照顾和妊娠压力引起的压力的遗传。第三, 星形胶质细胞和神经元之间基于乳酸的信号传递是星形胶质细胞和神经元之间沟通的重要方式 这些类型的细胞在学习和记忆中发挥作用，并在后代中受到产前应激的干扰。第四, 人类和啮齿动物PFC发育的改变是由于贫困的照顾和 妊娠压力会使后代成年后的行为脱轨。在这样的背景下，我们 假设小鼠精子中包含的RNA启动的灭绝学习缺陷 暴露在压力下，部分原因是IL-PFC中GR的可获得性改变，扰乱了基于乳酸的活动 IL-PFC神经元和未成熟的成人IL-PFC。为了验证这一假设，我们将使用生物化学， 分子遗传学、发育生物学和神经元活动的体内调控，重点是IL-2 应激雄性小鼠精子RNA进入胚胎所产生的动物的PFC 注射过的。通过针对细胞和区域的研究，我们的工作将提供关于压力如何- 精子RNA的诱导改变被翻译成神经生物学遗产，并可能具有翻译 通过确定可以作为治疗靶点的生物学来减轻此类遗产的负担，从而产生影响。

PROJECT SUMMARY: Famines, the Holocaust and other natural and anthropogenic events are providing evidence that the effects of trauma and stress extend beyond the ancestral generation and affect mental health in offspring. Remedying parental behavior that is perturbed by stress and mitigating stress during pregnancy have received attention for their utility in halting such legacies of stress. In contrast, less is known about how to halt legacies of paternal stress that occurred prior to conception of the affected offspring. To fill this gap in knowledge, we must first understand how stress-induced alterations in paternal sperm perturb neurobiology and derail mental health. With this intent, our goal is to determine how cell-type specific offspring neurobiology is impacted by stress-induced alterations in sperm RNA that have emerged as one mechanism via which paternal lineages bequeath legacies of stress to offspring. To achieve this goal, we rely on our experience studying legacies of paternal stress, learning and memory in mice and build on unpublished data demonstrating that injections of RNA from sperm of male mice exposed to stress into single cell zygotes resulted in deficits in extinction learning in adulthood. To begin our investigation into the neurobiological mechanisms that might underlie these deficits in extinction learning being set into motion by RNA in sperm exposed to stress, we propose a focus on glucocorticoid receptors (GRs) in the infra-limbic prefrontal cortex (IL-PFC), lactate-based activity of neurons in the IL-PFC, and development of the IL-PFC. Our focus is shaped by the following background. First, the IL-PFC is important for extinction learning. Second, epigenetic-based regulation of the GR gene has received the most attention in studies that have investigated intergenerational legacies of stress arising from abusive care-giving and gestational stress, in both humans and rodents. Third, lactate-based signaling between astrocytes and neurons is an important mode of communication between these cell types, plays a role in learning and memory, and is perturbed in offspring by ante-natal stress. Fourth, altered development of the PFC in humans and rodents as a consequence of impoverished caregiving and gestational stress derails behavior in offspring during adulthood. Motivated by this background, we hypothesize that deficits in extinction learning that are set into motion by RNA contained in sperm of mice exposed to stress result in part, from altered GR availability in the IL-PFC, disrupted lactate-based activity of IL-PFC neurons, and an immaturity of the adult IL-PFC. To test this hypothesis, we will use biochemistry, molecular genetics, developmental biology and in vivo manipulation of neuronal activity with a focus on the IL- PFC of animals generated from embryos into which RNA from sperm of male mice exposed to stress had been injected. Via cell- and region-specific investigations, our work will provide new insights into how stress- induced alterations in sperm RNA are translated into neurobiological legacies and may have translational impact by identifying biology that could be therapeutically targeted to lighten the burden of such legacies.