Neurosecretory Gene Expression in the Hypothalamus

下丘脑的神经分泌基因表达

• 批准号: 10239241
• 负责人: PAMELA L MELLON
• 金额: $ 54.05万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10239241
• 关键词: Address; Adult; Anterior; Apoptosis; Argipressin; Axon; Cell Differentiation process; Cell Line; Cells; Circadian Rhythms; Cues; Defect; Development; Embryo; Estrous Cycle; Exhibits; Failure; Feeds; Fertility; Foundations; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Goals; Gonadal Steroid Hormones; Gonadotropin Hormone Releasing Hormone; Heterozygote; Homeobox; Homeodomain Proteins; Hormones; Hypothalamic structure; In Vitro; Infertility; KISS1 gene; Knock-out; Knockout Mice; Laboratories; Methodology; Molecular; Morphology; Mus; Mutation; Neurites; Neuroendocrine Cell; Neurons; Neuropeptides; Neurosecretory Systems; Olfactory Nerve; Pathway interactions; Perinatal mortality demographics; Physiological; Pituitary Gland; Population; Puberty; Regulation; Regulatory Pathway; Reproduction; Role; Sex Behavior; Signal Transduction; Small Interfering RNA; Testosterone; Transfection; Transgenic Organisms; VIP gene; Vasoactive Intestinal Peptide; Ventral Anterior Thalamic Nucleus; circadian; circadian pacemaker; experience; homeodomain; in vitro Model; in vivo; innovation; insight; male; median eminence; migration; neural circuit; neuron development; neuronal circuitry; novel; promoter; reproductive; reproductive function; reproductive hormone; response; subfertility; suprachiasmatic nucleus; transcription factor; Address; Adult; Anterior; Apoptosis; Argipressin; Axon; Cell Differentiation process; Cell Line; Cells; Circadian Rhythms; Cues; Defect; Development; Embryo; Estrous Cycle; Exhibits; Failure; Feeds; Fertility; Foundations; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Goals; Gonadal Steroid Hormones; Gonadotropin Hormone Releasing Hormone; Heterozygote; Homeobox; Homeodomain Proteins; Hormones; Hypothalamic structure; In Vitro; Infertility; KISS1 gene; Knock-out; Knockout Mice; Laboratories; Methodology; Molecular; Morphology; Mus; Mutation; Neurites; Neuroendocrine Cell; Neurons; Neuropeptides; Neurosecretory Systems; Olfactory Nerve; Pathway interactions; Perinatal mortality demographics; Physiological; Pituitary Gland; Population; Puberty; Regulation; Regulatory Pathway; Reproduction; Role; Sex Behavior; Signal Transduction; Small Interfering RNA; Testosterone; Transfection; Transgenic Organisms; VIP gene; Vasoactive Intestinal Peptide; Ventral Anterior Thalamic Nucleus; circadian; circadian pacemaker; experience; homeodomain; in vitro Model; in vivo; innovation; insight; male; median eminence; migration; neural circuit; neuron development; neuronal circuitry; novel; promoter; reproductive; reproductive function; reproductive hormone; response; subfertility; suprachiasmatic nucleus; transcription factor

Project Summary The neural circuit that controls mammalian reproduction delivers circadian information from the suprachiasmatic nucleus (SCN) via the neuropeptides arginine vasopressin (AVP) and vasoactive intestinal peptide (VIP) to the kisspeptin (Kiss) and gonadotropin-releasing hormone (GnRH) neurons, respectively. Infertility can be caused at the neuroendocrine level by genetic defects in GnRH neurons, Kiss neurons, and/or circadian pacemaker and neuropeptide neurons in the SCN. GnRH neurons arise in the olfactory placode, migrate into the hypothalamus, and extend their axons to the median eminence to deliver GnRH to the pituitary in response to Kiss signals. Disrupted migration, neurite pathfinding, and/or disregulated secretion of Kiss or GnRH neurons result in failures of puberty, fertility, and reproductive function. We have determined that the homeodomain transcription factors, Vax1, Six3, and Six6, are all crucial for reproduction. Six6 Knock-out (KO) mice exhibit strikingly decreased fertility, while heterozygote Vax1 and Six3 mice are subfertile (Vax1 and Six3 KO mice are perinatal lethal). Though GnRH neuron numbers are normal at e13.5, all three of the KO embryos have ~90% reductions in hypothalamic GnRH neurons by e17.5. Vax1, Six3, and Six6 are found in Kiss neurons and regulate expression of the Kiss promoter in Kiss-expressing cell lines. In addition, deletion of Six3 from Kiss neurons causes subfertility. Furthermore, all three KO e17.5 embryos lack SCN morphology and fail to express AVP and VIP, and Six6 KO adults lack circadian rhythms. The overall goal of this application is to elucidate the development of this fundamental reproductive neuroendocrine circuit (SCNKissGnRH) at the molecular, developmental, and physiological levels. We propose three Specific Aims: Aim 1 will investigate differentiation and maturation of GnRH neurons during migration by determining the fate of neurons lacking these factors and effects on their migratory pathway. Aim 2 will focus on the roles of Vax1, Six3, and Six6 in the development and function of Kiss neurons in vitro and in vivo with a focus on direct regulation of Kiss gene expression and effects on fertility. Aim 3 will address the fundamental mechanisms of SCN development, the contribution of Vax1, Six3, and Six6 to AVP and VIP gene expression, and investigate the roles of SCN circadian function in fertility. Our overarching hypothesis is that infertility due to defects in Vax1, Six3, and Six6 is caused by alterations in specific hypothalamic neuroendocrine cell populations within the hypothalamic SCNKissGnRH reproductive neurocircuit by regulation of development, hormone synthesis, and circadian rhythms. We believe that studies of these key homeodomain proteins will reveal fundamental mechanisms in the control of reproductive function and circadian rhythms and provide valuable insight into novel regulatory pathways important for the development and function of this neuroendocrine circuit. This multifaceted approach will yield a comprehensive understanding of their roles in fertility and circadian rhythms in vivo and in vitro, and provide further insight into the fundamental genetic control of infertility and circadian rhythms.

项目摘要 控制哺乳动物繁殖的神经电路可从肩部提供昼夜节律信息 细胞核（SCN）通过神经肽精氨酸加压素（AVP）和血管活性肠肽（VIP）到 亲吻肽（KISS）和促性腺激素释放激素（GNRH）神经元。不育可能引起 在神经内分泌水平上，GNRH神经元中的遗传缺陷，亲吻神经元和/或昼夜节律起搏器以及 SCN中的神经肽神经元。 gnRH神经元在嗅觉上出现，迁移到下丘脑中， 并将其轴突扩展到中间的隆起，以响应亲吻信号，将gnrh传递到垂体。 破坏的迁移，神经落路线和/或无视亲吻或GNRH神经元的分泌导致失败 青春期，生育能力和生殖功能。我们已经确定同源域转录因子， VAX1，SIX3和SIX6对于繁殖至关重要。 Six6敲除（KO）小鼠暴露了明显减少 生育能力，而杂合子VAX1和63只小鼠是属的（VAX1和Six3 KO小鼠是围产期致死的）。 尽管GnRH神经元数在E13.5时正常，但所有三个KO胚胎均降低了约90％ E17.5的下丘脑GNRH神经元。 VAX1，SIX3和SIX6在Kiss神经元中发现并调节表达 用表达亲吻的细胞系中的亲吻启动子的摄影。此外，从KISS神经元原因中删除了六3 亚属。此外，所有三个KO E17.5胚胎都缺乏SCN形态，并且无法表达AVP和VIP， 六个KO成年人缺乏昼夜节律。该应用程序的总体目标是阐明开发 在分子，发育， 和身体水平。我们提出了三个具体目标：AIM 1将研究分化和成熟 通过确定缺乏这些因素的神经元的命运和对其影响的影响，GNRH神经元的gnRH神经元的命运 迁移途径。 AIM 2将重点关注VAX1，SIX3和SIX6在KISS的发展和功能中的作用 在体外和体内神经元，重点是直接调节亲吻基因表达和对生育能力的影响。目的 3将解决SCN开发的基本机制，VAX1，SIX3和SIX6的贡献 AVP和VIP基因表达，并研究SCN昼夜节律在生育中的作用。我们的总体 假设是，由于特定的变化，由于VAX1，SIX3和SIX6中缺陷引起的不育症是引起的 下丘脑的下丘脑神经内分泌细胞群体中的SCNkiss genrh生殖 通过调节发育，马-mon合成和昼夜节律来调节神经信仰。我们认为研究 这些关键的同源域蛋白将揭示复制功能控制中的基本机制 和昼夜节律，并为新的监管途径提供了宝贵的见解 和该神经内分泌电路的功能。这种多方面的方法将产生全面的理解 它们在体内和体外的生育和昼夜节律中的作用，并进一步了解基本 不育和昼夜节律的遗传控制。