Estrogen regulation of channels involved in the control of energy homeostasis

描述（由申请人提供）： 本申请的长期目标是了解雌激素和环境雌激素影响下丘脑功能如能量稳态的机制。在美国存在肥胖症流行病，并且了解导致肥胖症表达的任何和所有因素的病因对于治疗这种疾病至关重要。因此，指导项目的目标是了解雌二醇如何通过多种膜启动机制影响能量稳态，包括通过雌二醇响应膜GPCR（mER）控制神经元兴奋性和通过ER？和mER介导的机制。众所周知，雌激素通过ER控制能量稳态？和mER，因为STX，mER的选择性配体，减弱卵巢切除术后的体重增加。雌二醇还通过mER改变POMC弓状神经元的神经元兴奋性。雌二醇作用的一种潜在mER介导机制是调节非失活亚阈值K+电流，该电流可调节POMC的兴奋性（M电流）。为了确定雌二醇对M-电流的调节是否在这些效应中起作用，我将首先在合并的单细胞和电生理学中使用qRT-PCR测量来自油和雌二醇处理的雌性的POMC和NPY神经元中KCNQ/M-电流的表达和活性。第二，我将测量急性（通过mER）雌二醇治疗对POMC和NPY神经元的M-电流的电生理特性的影响，使用全细胞膜片记录。独立阶段将建立在指导阶段学习的技术基础上，并研究膜启动和雌激素非依赖性信号传导在下丘脑功能中的作用，并确定环境雌激素是否激活这些相同的通路。第一个目标将确定的作用，雌激素受体的非依赖性信号在控制能量稳态和下丘脑的基因调控雌二醇和双酚A使用野生型，？ERKO和ER α KI/KO小鼠模型。最终的目标是确定双酚A对弓形POMC和NPY神经元控制能量稳态的电生理效应。 公共卫生相关性：指导阶段可能直接影响妇女（尤其是绝经后妇女）的健康，因为STX是膜雌激素受体的选择性配体，是非传统激素替代疗法和治疗肥胖症的潜在先导化合物。最后，独立阶段将解决环境雌激素可能影响下丘脑功能的潜在机制。

DESCRIPTION (provided by applicant): The long range goal of this application is to understand the mechanisms by which estrogen and environmental estrogens affect hypothalamic functions such as energy homeostasis. There is an obesity epidemic in the US and understanding the etiology of any and all factors that contribute to expression of obesity is critical for treatment of this disorder. Therefore, the goal of the mentored projects is to understand how estradiol affects energy homeostasis through multiple membrane-initiated mechanisms that include the control of neuronal excitability through an estradiol-responsive membrane GPCR (mER) and control of gene expression in arcuate neurons through both ER?- and mER-mediated mechanisms. Estradiol is known to control energy homeostasis through ER? and mER because STX, a selective ligand for the mER, attenuates body weight gain post-ovariectomy. Estradiol also alters neuronal excitability of POMC arcuate neurons through the mER. One potential mER-mediated mechanism for the effects of estradiol is modulation of a non-inactivating, sub-threshold K+ current that tempers the excitability of POMC (M-current). To determine if modulation of the M-current by estradiol plays a role in these effects, I will first measure the expression and activity of the KCNQ/M-current in POMC and NPY neurons from oil- and estradiol-treated females using qRT-PCR in pooled single cells and electrophysiology. Second, I will measure the effects of acute (via mER) estradiol treatment on the electrophysiological properties of the M-current in POMC and NPY neurons using whole cell patch recordings. The independent phase will build on the techniques learned during the mentored phase and examine the role of membrane-initiated and ERE-independent estrogen signaling in hypothalamic functions and determine whether environmental estrogens activate these same pathways in their effects. The first aim will determine the role of ERE-independent signaling in the control of energy homeostasis and hypothalamic gene regulation by estradiol and bisphenol A using wild-type, ?ERKO and ERalpha KI/KO mice models. The final aim will determine the electrophysiological effects of bisphenol A on arcuate POMC and NPY neurons that control energy homeostasis. PUBLIC HEALTH RELEVANCE: The mentored phase may directly impact the health of women (esp. post-menopausal women) since STX, the selective ligand for the membrane estrogen receptor, is a potential lead compound for an non-traditional hormone replacement therapy and treatment of obsesity disorders. Finally, the independent phase will address the potential mechanisms that environmental estrogens can impact hypothalamic functions.