Estradiol modulation of pacemaking kisspeptin neurons

雌二醇对起搏 Kisspeptin 神经元的调节

基本信息

批准号：
9096268
负责人：
Oline Karin Rønnekleiv
金额：
$ 42.68万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-01-01 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9096268
关键词：
Address Adult Animals Appetite Stimulants Area Calcium Cells Central Nervous System Diseases Characteristics Development Eating Electrophysiology (science)Estradiol Exhibits Female Gene Expression Generations Goals Health Histocytochemistry Homeostasis Hormones Hypothalamic structure Ion Channel KISS1 gene KISS1R gene Light Longevity Mediating Metabolic Diseases Molecular Molecular Biology Molecular Profiling Neurons Neuropathy Pacemakers Peptides Physiology Pro-Opiomelanocortin Process Regulation Reproduction Research Reverse Transcriptase Polymerase Chain Reaction Rhodopsin Risk Signal Transduction Signaling Protein Sodium Channel Stroke Structure of nucleus infundibularis hypothalami Synapses Third ventricle structure Translating Whole-Cell Recordings Woman base biophysical properties energy balance estrogenic feeding in vivo inhibitory neuron insight interdisciplinary approach mRNA Expression neural circuit neuronal circuitry neuropeptide Y novel novel therapeutics optogenetics postsynaptic receptor research study response tool treatment strategy

项目摘要

DESCRIPTION (provided by applicant): The long-range goals of the proposed research are to elucidate the molecular and cellular mechanisms by which 17ß-estradiol (E2) regulates hypothalamic kisspeptin (Kiss1) neuronal circuits in females. Kiss1 neurons are a major target of E2 and are essential for pubertal development and adult reproduction. These neurons are also important for sensing and relaying gonadal status to neurons vital for regulation of energy balance, including the hypothalamic arcuate proopiomelanocortin (POMC) and neuropeptide Y/agouti-related peptide (NPY/AgRP) neurons. In spite of the importance of Kiss1 neurons, relatively little is known about the biophysical properties and molecular signature of these cells, as well as the neural circuits by which they operate. We have discovered that the Kiss1 neurons in the rostral periventricular area of the third ventricle (RP3V) express the critical ion channels and receptors that permit spontaneous E2-dependent rebound burst firing. Based on the E2-dependent burst firing characteristics and their projections to arcuate neurons, we propose the novel hypothesis that the RP3V Kiss1 neurons serve as the central pacemaker neurons that provide E2-dependent excitatory drive to the anorexigenic POMC neurons and inhibitory drive to the orexigenic NPY/AgRP neurons. This kisspeptin input to POMC and NPY/AgRP neurons is vital for the estrogenic control of energy homeostasis during the ovulatory cycle in females. Elucidating the cell-specific signaling pathways and gene expression at the single cell level will help in developing new strategies for targeting hormone actions in CNS neurons. Our multidisciplinary approach incorporates a unique set of cellular, molecular, optogenetic and chemogenetic tools and our combined expertise in molecular biology, electrophysiology, histochemistry and whole animal physiology to address the following aims: 1) To elucidate the sodium channel (Nav) subunit composition and the contribution of INaP to the generation of burst firing in RP3V Kiss1 neurons in high- versus low-E2 states. 2) To elucidate the postsynaptic actions of kisspeptin on arcuate POMC and NPY/AgRP neurons using single cell RT-PCR and whole-cell recording. 3) To elucidate the direct synaptic input to arcuate POMC and NPY/AgRP neurons from RP3V Kiss1 neurons using optogenetic stimulation and whole-cell recording. 4) To examine the in vivo effects of selective chemogenetic activation or inhibition of RP3V Kiss1 neurons on food intake in low- versus high-E2 females, respectively. Collectively, these experiments will allow for the elucidation of the ion channels important for the pacemaker activity in hypothalamic Kiss1 neurons, and how this translates into actions within hypothalamic neurocircuits that are vital for not only energy homeostasis but other autonomic functions.

描述（由申请人提供）：拟议研究的长期目标是阐明 17β-雌二醇 (E2) 调节女性下丘脑 Kisspeptin (Kiss1) 神经元回路的分子和细胞机制，而 Kiss1 神经元是 Kiss1 神经元的主要目标。 E2 对青春期发育和成年生殖至关重要，这些神经元对于感知性腺状态并将其传递给对能量平衡调节至关重要的神经元也很重要，包括下丘脑弓状阿片黑皮素原 (POMC) 和神经肽 Y/agouti 相关肽 (NPY/AgRP) 神经元尽管 Kiss1 神经元很重要，但人们对这些细胞的生物物理特性和分子特征知之甚少。以及它们运作的神经回路我们发现第三脑室（RP3V）头侧脑室周围区域的 Kiss1 神经元表达关键离子通道。以及允许自发 E2 依赖性反弹爆发放电的受体基于 E2 依赖性爆发放电特性及其对弓形神经元的投射，我们提出了新的假设：RP3V Kiss1 神经元充当提供 E2 依赖性兴奋性的中央起搏神经元。 Kisspeptin 对 POMC 和 NPY/AgRP 神经元的雌激素控制至关重要。阐明女性排卵周期期间的能量稳态，将有助于开发针对中枢神经系统神经元激素作用的新策略。光遗传学和化学遗传学工具以及我们在分子生物学、电生理学、组织化学和整个动物生理学方面的综合专业知识，旨在实现以下目标：1) 阐明钠通道 (Nav) 亚基的组成和贡献INaP 与高 E2 状态和低 E2 状态下 RP3V Kiss1 神经元中爆发放电的产生有关 2) 使用单细胞 RT-PCR 和全细胞记录阐明 Kisspeptin 对弓形 POMC 和 NPY/AgRP 神经元的突触后作用。 ) 使用光遗传学刺激和全细胞阐明从 RP3V Kiss1 神经元到弓形 POMC 和 NPY/AgRP 神经元的直接突触输入4）为了检查 RP3V Kiss1 神经元的选择性化学遗传学激活或抑制分别对低 E2 雌性和高 E2 雌性的食物摄入的体内影响，这些实验将有助于阐明对 RP3V Kiss1 神经元重要的离子通道。下丘脑 Kiss1 神经元中的起搏器活动，以及如何将其转化为下丘脑神经回路内的活动，这些活动不仅对于能量稳态而且对于其他自主功能都至关重要。