Determining the role of RFRP-3 in stress-related female infertility

描述（由申请人提供）：拟议的项目是一个调查压力和不孕症之间的潜在机制联系的妇女。女性生殖系统可能更容易出现压力诱导的失调，因为女性周期需要精确和平衡的激素信号沿着下丘脑-垂体-性腺（HPG）轴。尽管如此，应激反应系统和生殖系统之间的机械联系尚未被定义。这项研究的目的是表征应激如何通过RFRP-3与HPG轴相互作用，RFRP-3是促性腺激素抑制激素的哺乳动物直系同源物，该激素已被证明直接抑制GnRH和LH释放。在拟议的项目中，我们将首先描述女性系统和RFRP-3的压力相关影响之间的相互作用。为此，本提案的第一个目标将定义在发情周期的每个阶段急性应激对RFRP-3的具体影响，同时还确定应激反应系统的哪些组分直接影响RFRP-3的表达。在此目标下，候选人将学习和利用激光捕获显微镜和实时RT-PCR分离和精确分析RFRP-3神经元上的相关糖皮质激素和促肾上腺皮质激素释放激素受体（GR和CRH-R1）。此外，候选人将学习利用病毒载体表达一种类型的 GR，破坏来自皮质酮（啮齿动物中的主要应激激素）的应激诱导信号，从而精确分析这种激素在RFRP-3神经元水平上的作用。第二个目标将研究慢性应激对系统的影响，研究人员将通过测量RFRP神经元对应激激素的敏感性变化和GnRH神经元对RFRP-3的敏感性变化，研究下丘脑水平的敏感性变化如何导致促性腺激素释放改变。虽然本项目主要侧重于女性系统，但在有关男性主体的信息不完整的情况下，将分析总体性别差异。总体预测是，RFRP-3在应激反应系统和生殖系统之间的联系中起着重要作用，并且在急性应激水平下，这种联系是快速和暂时的。然而，在暴露于慢性应激期间和之后，预测RFRP-3释放的调节和对激素的敏感性被破坏，使得应激系统和生殖系统之间的这种信号导致HPG轴失调和下游效应，其可导致生育力降低。最终，这项研究的目标是帮助确定一种失调模式的压力诱导不孕症的妇女。

DESCRIPTION (provided by applicant): The proposed project is an investigation into a potential mechanistic link between stress and infertility in women. The female reproductive system may be more prone to stress-induced dysregulation since female cycling requires precise and balanced hormone signals along the hypothalamic-pituitary-gonadal (HPG) axis. Despite this, the mechanistic link between the stress response system and the reproductive system has yet to be defined. This research aims to characterize how stress interacts with the HPG axis via RFRP-3, the mammalian ortholog for gonadotropin inhibitory hormone, a hormone that has been shown to directly suppress GnRH and LH release. In the proposed project, we will first characterize the interplay between the female system and stress-related effects on RFRP-3. To this end, the first aim of this proposal will define specific effects of acute stress on RFRP3 at each stage of the estrous cycle while also determining which components of the stress response system directly affect RFRP-3 expression. Within this aim, the candidate will learn and utilize laser capture microscopy and real-time RT-PCR to isolate and precisely analyze relative glucocorticoid and corticotropin releasing hormone receptors (GR and CRH-R1, respectively) on RFRP-3 neurons. In addition, the candidate will learn to utilize viral vectors to express a type of GR that disrupts the stress-induced signal from corticosterone (the main stress hormone in rodents) thus precisely analyzing the role of this hormone at the level of RFRP-3 neurons. The second aim will investigate chronic stress effects on the system and the researchers will investigate how changes in sensitivity at the level of the hypothalamus may lead to altered gonadotropin release by measuring the change in sensitivity of RFRP neurons to stress hormones and the change in sensitivity of GnRH neurons to RFRP-3. While this project mainly focuses on the female system overall sex differences will be analyzed when information on male subjects is incomplete. The overall prediction is that RFRP-3 plays an important role in the link between the stress response system and the reproductive system and that, at the acute stress level, this link is rapid and temporary. However, during and following exposure to chronic stress, the regulation of RFRP-3 release and sensitivity to the hormone is predicted to be disrupted such that this signal between the stress system and the reproductive system results in a dysregulated HPG axis and downstream effects that can lead to decreased fertility. Ultimately, the goal of this research is to help define a mode of dysregulation underlying stress-induced infertility in women.