The Kiss1 system in the neuroendocrine control of reproduction

Kiss1系统在生殖神经内分泌控制中的作用

• 批准号: 8280018
• 负责人: Victor Manuel Navarro
• 金额: $ 13.72万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-12-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8280018
• 关键词: Affect; Animal Model; Animals; Bilateral; Breeding; Castration; Cell Nucleus; Clinical Investigator; Complement; Contraceptive methods; Couples; Development; Disease; Dynorphin A; Dynorphins; Estradiol; Estrus; Exhibits; Family Planning; Feedback; Fellowship; Female; Generations; Genes; Genetic; Genetic Techniques; Genetically Engineered Mouse; Goals; Gonadal Steroid Hormones; Gonadotropin Hormone Releasing Hormone; Gonadotropins; Hospitals; Human; Hypothalamic structure; Impairment; Infertility; Injection of therapeutic agent; KISS1R gene; Klinefelter' s Syndrome; Knowledge; Lead; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Mentors; Methodology; Methods; Modeling; Molecular; Molecular Genetics; Mus; Mutation; Neurokinin B; Neurons; Neurosecretory Systems; Outcome Study; Ovarian; Ovulation Induction; Pathway interactions; Periodicity; Phase; Physiologic pulse; Physiological; Population; Postdoctoral Fellow; Precocious Puberty; Principal Investigator; Progesterone; Puberty; Recurrence; Regulation; Reproduction; Research; Research Personnel; Research Proposals; Rodent; Role; Shapes; Signal Transduction; Source; Structure of nucleus infundibularis hypothalami; System; Techniques; Time; Training; Viral; Weight; Woman; adeno-associated viral vector; career; design; experience; gene therapy; hormone therapy; improved; kisspeptin; loss of function mutation; male; mouse model; novel strategies; prepuberty; programs; promoter; public health relevance; receptor; recombinase; reproductive; reproductive axis; reproductive development; reproductive function; reproductive neuroendocrinology; research study; skills; success; treatment strategy

DESCRIPTION (provided by applicant): This K99/R00 proposal is designed to complement and advance my prior research experience. Briefly, my expertise is in the study of the neuronal mechanisms that modulate reproductive function. I have contributed to the characterization of the roles of kisspeptins (encoded by the Kiss1 gene) as regulatory conduits of gonadotropin-releasing hormone (GnRH) release. During my post-doctoral fellowship, I have broadened this research to include the central modulators of kisspeptin release, neurokinin B (NKB) and dynorphin A (Dyn), which are co-expressed with kisspeptin in the arcuate nucleus (ARC). We have proposed a model whereby NKB and Dyn act through recurrent collaterals to shape the pulsatile release of kisspeptin that would ultimately lead to pulses in GnRH release. The overall goal of this research proposal is to understand the physiological function of subpopulations of Kiss1 neurons that regulate the secretion of GnRH and hence reproduction. Kiss1 expressing neurons are found in two discrete nuclei in the hypothalamus, the ARC and anteroventral periventricular nucleus (AVPV), and are direct targets for the negative and positive feedback action of estradiol E2 on GnRH secretion, respectively. Kiss1 expression in these neurons increases at the time of puberty onset and humans and rodents bearing inactivating mutations in this gene or its receptor (Kiss1r) exhibit impuberism and infertility. The objective of this proposal is to use genetic mouse models in combination with viral-dependent gene therapy to dissect the differential role of the ARC and AVPV Kiss1 and NKB neuronal populations in the control of reproduction. We will use genetically engineered mice that express Cre recombinase and GFP under the Kiss1 promoter. Breeding to homozygosity would lead to Kiss1 deficient (but Cre/GFP positive) mice. Using Cre-dependent adeno-associated viral constructs, we aim to reinsert the Kiss1 gene specifically in Kiss1 neurons of the ARC or the AVPV. The specific aims of the project are to: 1) determine the role of Kiss1 neurons in the ARC vs. AVPV in the control of puberty onset in male and female mice; 2) determine the role of Kiss1 neurons in the ARC vs. AVPV in the positive and negative feedback of sex steroids; and 3) determine the role of NKB in the ARC in the control of puberty in male and female mice. The studies outlined in this proposal are designed to advance our fundamental knowledge of reproductive neuroendocrinology, for the development of better strategies to treat reproductive disorders and offer improved methods of contraception. Completion of these studies will provide training in: (a) state-of-the-art geneti and molecular methodologies; (b) addition of a translational component to my research studies through close interaction with clinical investigators; and (c) skills necessary for success as an independent principal investigator. This additional training will aid in my development of a competitive research program and contribute to my establishment as an independent investigator.

描述（由申请人提供）：本 K99/R00 提案旨在补充和推进我之前的研究经验。简而言之，我的专长是研究调节生殖功能的神经元机制。我对 Kisspeptins（由 Kiss1 基因编码）作为促性腺激素释放激素 (GnRH) 释放调节管道的作用的表征做出了贡献。在我的博士后研究期间，我扩大了这项研究范围，将 Kisspeptin 释放的中枢调节剂包括神经激肽 B (NKB) 和强啡肽 A (Dyn)，它们与弓状核 (ARC) 中的 Kisspeptin 共同表达。我们提出了一个模型，NKB 和 Dyn 通过循环循环发挥作用，形成 Kisspeptin 的脉冲式释放，最终导致 GnRH 释放的脉冲。这项研究计划的总体目标是了解 Kiss1 神经元亚群的生理功能，这些神经元调节 GnRH 的分泌，从而调节生殖。 Kiss1 表达神经元存在于下丘脑的两个离散核中，即 ARC 和前腹室周围核 (AVPV)，它们分别是雌二醇 E2 对 GnRH 分泌负反馈和正反馈作用的直接靶标。这些神经元中 Kiss1 的表达在青春期开始时增加，携带该基因或其受体 (Kiss1r) 失活突变的人类和啮齿动物表现出青春期不成熟和不育。该提案的目的是使用遗传小鼠模型与病毒依赖性基因治疗相结合来剖析 ARC 和 AVPV Kiss1 以及 NKB 神经元群在控制繁殖中的不同作用。我们将使用在 Kiss1 启动子下表达 Cre 重组酶和 GFP 的基因工程小鼠。纯合性繁殖将产生 Kiss1 缺陷（但 Cre/GFP 阳性）小鼠。使用 Cre 依赖性腺相关病毒构建体，我们的目标是将 Kiss1 基因特异地重新插入 ARC 或 AVPV 的 Kiss1 神经元中。该项目的具体目标是： 1) 确定 ARC 与 AVPV 中 Kiss1 神经元在控制雄性和雌性小鼠青春期开始方面的作用； 2）确定ARC vs. AVPV中Kiss1神经元在性类固醇正反馈和负反馈中的作用； 3) 确定 ARC 中的 NKB 在控制雄性和雌性小鼠青春期中的作用。该提案中概述的研究旨在增进我们对生殖神经内分泌学的基础知识，以制定更好的策略来治疗生殖疾病并提供改进的避孕方法。完成这些研究将提供以下方面的培训： (a) 最先进的遗传和分子方法学； (b) 通过与临床研究人员的密切互动，为我的研究添加转化成分； (c) 作为独立首席研究员取得成功所需的技能。这些额外的培训将有助于我制定有竞争力的研究计划，并有助于我成为一名独立调查员。