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

描述（由申请人提供）：拟议的项目是对压力与女性不孕之间潜在机制联系的调查。女性生殖系统可能更容易出现压力引起的失调，因为女性周期需要沿着下丘脑-垂体-性腺（HPG）轴精确和平衡的激素信号。尽管如此，应激反应系统和生殖系统之间的机制联系尚未确定。本研究旨在通过RFRP-3表征应激如何与HPG轴相互作用，RFRP-3是促性腺激素抑制激素的哺乳动物同源物，一种被证明直接抑制GnRH和LH释放的激素。在拟议的项目中，我们将首先描述女性系统与压力相关的RFRP-3影响之间的相互作用。为此，本提案的第一个目标将确定在发情周期的每个阶段急性应激对RFRP3的具体影响，同时确定应激反应系统的哪些组成部分直接影响RFRP3的表达。在此目标下，候选人将学习并利用激光捕获显微镜和实时RT-PCR分离和精确分析RFRP-3神经元上的糖皮质激素和促肾上腺皮质激素释放激素受体（分别为GR和CRH-R1）。此外，候选人将学习利用病毒载体来表达一种

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.