The role of kisspeptin and RFRP-3 in the neuroendocrine control of female reproduction

项目摘要 成功的雌性生殖需要大量的 神经内分泌事件的主昼夜节律起搏器在视交叉上核（SCN）。跨 物种，包括人类，昼夜节律的中断导致明显的异常， 发情/月经周期，生育能力下降，流产率增加。我们迄今为止的发现提供了 有证据表明，在一个排卵控制网络中，SCN在时间上协调两个关键的 具有相反作用的神经肽能系统，RFamide相关肽-3（RFRP-3，哺乳动物神经肽能系统）， 禽类促性腺激素抑制激素的直系同源物）和kisspeptin系统。在哺乳动物中， RFRP-3和kisspeptin分别是生殖轴的关键抑制和刺激调节剂。 尽管这些神经肽在哺乳动物生殖中的作用已经确立， 知道昼夜节律计时系统是女性生殖轴的重要调节器， 这些系统之间的相互作用可以适当地协调神经内分泌的时间。 排卵所必需的事件仍未明确。为了了解这个网络的元素如何协同作用， 产生排卵所需的促性腺激素释放激素（GnRH）系统的协调输出， 本发明建议使用转基因小鼠模型，结合细胞特异性病毒方法， 阐明生物钟运作的细胞机制和神经化学信号通路 与RFRP-3和Kp系统接口。具体而言，本建议将：1）确定 SCN通过其与RFRP-3和kisspeptin系统通信的神经化学信号传导途径， 2)系统地检查已鉴定途径的功能贡献，并具有细胞特异性精度， 和3）确定自主神经内分泌细胞昼夜节律计时在此过程中的功能意义。 通过细胞表型特异性敲低基本时钟基因的控制网络。拟议的工作不 不仅解决了生殖生物学中的一个经典问题，而且还具有实质性的翻译潜力。 在发展安全/有效避孕以及治疗一系列生殖疾病方面的影响 人类的疾病，包括性早熟和青春期延迟、不育和多囊卵巢综合征。

Project Summary Successful female reproduction requires the precise temporal coordination of numerous neuroendocrine events by a master circadian pacemaker in the suprachiasmatic nucleus (SCN). Across species, including humans, disruptions to circadian timing result in pronounced abnormalities in the estrous/menstrual cycle, reductions in fertility, and increased miscarriage rates. Our findings to date provide evidence for a network of ovulatory control in which the SCN temporally coordinates the activity of two, key neuropeptidergic systems with opposing actions, the RFamide-related peptide-3 (RFRP-3, the mammalian ortholog of avian gonadotropin-inhibitory hormone) and kisspeptin systems. Across mammalian species, RFRP-3 and kisspeptin are key inhibitory and stimulatory regulators of the reproductive axis, respectively. Despite the well-established role for these neuropeptides in mammalian reproduction, as well as the knowledge that the circadian timing system is a crucial regulator of the female reproductive axis, the specific means by which interactions among these systems appropriately coordinate the timing of neuroendocrine events necessary for ovulation remain unspecified. To understand how elements of this network synergize to produce the coordinated output to the gonadotropin-releasing hormone (GnRH) system required for ovulation, the present proposal uses transgenic mouse models, in combination with cell-specific viral approaches, to elucidate the cellular mechanisms and neurochemical signaling pathways by which the circadian clockwork interfaces with the RFRP-3 and Kp systems. Specifically, the present proposal will: 1) identify the neurochemical signaling pathways by which the SCN communicates with the RFRP-3 and kisspeptin systems, 2) examine the functional contribution of identified pathways systematically, and with cell-specific precision, and 3) determine the functional significance of autonomous, neuroendocrine-cell circadian timekeeping in this network of control through cell-phenotype-specific knockdown of essential clock genes. The proposed work not only addresses a classic question in reproductive biology, but also has the potential for substantial translational impact in the development of safe/effective contraception, as well as the treatment of a host of reproductive disorders in humans, including precocious and delayed puberty, infertility, and polycystic ovarian syndrome.