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）：此K99/R00建议旨在补充和推进我先前的研究经验。简而言之，我的专业知识是研究调节生殖功能的神经元机制。我为促促性腺激素释放激素（GNRH）释放的吻肽（由Kiss1基因编码）的角色（由Kiss1基因编码）的角色做出了贡献。在博士后研究金期间，我扩大了这项研究，包括Kisspeptin释放的中心调节剂，Neurokinin B（NKB）和Dynorphin A（Dyn）A（Dyn），它们在弧形核（ARC）中与Kisspeptin共表达。我们提出了一个模型，NKB和DYN通过反复的侧边起作用，以塑造亲吻肽的脉冲释放，最终导致GNRH释放中的脉冲。该研究建议的总体目标是了解调节GNRH分泌并因此再现的Kiss1神经元亚群的生理功能。在下丘脑，弧和前脑室脑周围核（AVPV）中，在两个离散核中发现了KISS1表达神经元，并且分别是雌二醇E2对GNRH分泌的负反馈作用的直接靶标。这些神经元中的KISS1表达在青春期发作时增加，并且人类和啮齿动物在该基因或其受体（KISS1R）中带有灭活突变的啮齿动物表现出无关的和不育。该建议的目的是将遗传小鼠模型与病毒依赖性基因治疗结合使用，以剖析ARC和AVPV KISS1和NKB神经元种群在繁殖控制中的差异作用。我们将使用基因工程的小鼠在Kiss1启动子下表达CRE重组酶和GFP。繁殖至纯合性会导致Kiss1缺乏（但CRE/GFP阳性）小鼠。使用依赖CRE的腺相关病毒构建体，我们旨在将Kiss1基因特别插入ARC或AVPV的Kiss1神经元中。该项目的具体目的是：1）确定KISS1神经元在ARC与AVPV中的作用在雄性和雌性小鼠的青春期发作中的作用； 2）确定KISS1神经元在ARC与AVPV中的作用在性类固醇的正面和负面反馈中； 3）确定NKB在弧度控制雄性和雌性小鼠中的作用。该提案中概述的研究旨在提高我们对生殖神经内分泌学的基本知识，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。这些研究的完成将提供：（a）最先进的基因和分子方法； （b）通过与临床研究者的密切相互作用，在我的研究中添加了翻译成分； （c）作为独立首席研究员成功所必需的技能。这种额外的培训将有助于我制定竞争性研究计划，并为我作为独立调查员的成立做出贡献。