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/R 00提案旨在补充和推进我以前的研究经验。简而言之，我的专长是研究调节生殖功能的神经机制。我对kisspeptins（由Kiss 1基因编码）作为促性腺激素释放激素（GnRH）释放的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，包括kisspeptin释放的中央调节剂，神经激肽B（NKB）和强啡肽A（Dyn），它们与弓状核（ARC）中的kisspeptin共表达。我们已经提出了一个模型，NKB和Dyn通过反复的侧支作用，形成kisspeptin的脉冲式释放，最终导致GnRH释放的脉冲。这项研究的总体目标是了解Kiss 1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。Kiss 1表达神经元被发现在两个离散的核在下丘脑，ARC和前腹侧室周核（AVPV），并分别是雌激素E2对GnRH分泌的负反馈和正反馈作用的直接目标。这些神经元中的Kiss 1表达在青春期开始时增加，并且在该基因或其受体（Kiss 1 r）中具有失活突变的人类和啮齿动物表现出青春期和不育。本提案的目的是使用遗传小鼠模型结合病毒依赖性基因治疗，以剖析ARC和AVPV Kiss 1和NKB神经元群体在控制生殖中的不同作用。我们将使用在Kiss 1启动子下表达Cre重组酶和GFP的基因工程小鼠。纯合性育种将导致Kiss 1缺陷（但Cre/GFP阳性）小鼠。使用依赖Cre的腺相关病毒构建体，我们的目标是重新插入Kiss 1基因，特别是在ARC或AVPV的Kiss 1神经元。该项目的具体目的是：1）确定ARC与AVPV中Kiss 1神经元在控制雄性和雌性小鼠青春期开始中的作用; 2）确定ARC与AVPV中Kiss 1神经元在性类固醇正反馈和负反馈中的作用; 3）确定ARC中NKB在控制雄性和雌性小鼠青春期中的作用。本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，以制定更好的治疗生殖疾病的战略，并提供更好的避孕方法。完成这些研究将提供以下方面的培训：（a）最先进的遗传学和分子方法学;（B）通过与临床研究者的密切互动，在我的研究中增加翻译部分;以及（c）作为独立主要研究者取得成功所需的技能。这一额外的培训将有助于我制定一个有竞争力的研究计划，并有助于我成为一名独立的研究者。