Membrane estrogen receptors and ingestive behavior

说明(申请人提供)：雌二醇(E2)在维持适当的能量和体液平衡方面起着至关重要的作用。E2的这一关键作用在绝经后妇女中尤其明显，她们的E2水平降低，患肥胖症、肥胖相关疾病和高血压的风险增加。不幸的是，E2调节能量和液体平衡的机制仍然知之甚少。啮齿动物研究表明，雌激素通过激活经典的雌激素受体亚型雌激素受体？(呃？？)。呃？？定位于细胞核，在那里它通过与DNA上的反应元件相互作用来改变基因表达。更新的发现是ER？？也可以与细胞膜结合并作为表面受体，导致人们认识到这种受体可以通过替代的、非传统的机制影响基因的表达，包括激活转录因子，如cAMP反应元件结合蛋白(CREB)。我们实验室最近的研究表明，膜相关雌激素受体(MER)的激活会减少食物和水的摄入量。因此，这项建议的目的是在液体摄入、细胞间信号传递和病毒介导的遗传操作领域提供培训，并增强候选人在雌激素调节食物摄入领域的现有优势。这将是 部分是通过进行拟议的实验来实现的，这些实验测试了压倒一切的假设，即Mer通过参与传统上与转录变化相关的细胞内信号通路来减少食物和水的摄入。为此，本方案中的实验包括两个目的，第一个目的是测试代谢性谷氨酸受体(MGluR)在调节Mer对食物和水摄取的影响中的需求。第二个目标将通过AAV操作检查MER被击倒后的食物和水摄入量。接下来，这组实验将测试MER？？之间的物理相互作用。和mGluR，并将测试mGluR对E2细胞内效应的作用。除了为候选人提供重要的培训，帮助她过渡到独立科学家的职业生涯之外，拟议中的研究还有可能增加对E2信号影响食物和水摄入量的机制的理解，并有可能揭示类固醇对基因表达影响的更一般原理。

DESCRIPTION (provided by applicant): Estradiol (E2) plays a crucial role in the maintenance of proper energy and fluid homeostasis. This key role of E2 is especially apparent in postmenopausal women who have decreased E2 levels and are at an elevated risk for obesity, obesity-related diseases, and hypertension. Unfortunately, the mechanism by which E2 regulates energy and fluid balance is still poorly understood. Rodent studies indicate that estrogens decrease food and water intake through a genomic mechanism via activation of the classical estrogen receptor subtype, estrogen receptor ? (ER??). ER?? localizes to the cell nucleus, where it alters gene expression by interacting with responsive elements on DNA. The more recent discovery that ER?? also can associate with the cell membrane and act as a surface receptor led to the appreciation that this receptor can affect gene expression through alternate, non-traditional mechanisms including activation of transcription factors such as cAMP response element binding protein (CREB). Recent studies from our laboratory demonstrate that activation of membrane-associated estrogen receptors (mER) decrease food and water intake. Accordingly, this proposal was designed to provide training in the areas of fluid intake, intercellular signaling and viral mediated genetic manipulations in addition to enhance the existing strengths of the candidate in the area of estrogen modulation of food intake. This will be accomplished, in part, by performing the proposed experiments that test the overarching hypothesis that mER decreases food and water intake by engaging intracellular signaling pathways traditionally associated with transcriptional changes. To this end, the experiments in this proposal include two aims, the first of which will test the requirement of metabotropic glutamate receptors (mGluR) in mediating the effects of mER on food and water intake. The second aim will examine food and water intake following knockdown of mER through AAV manipulation. Next this set of experiments will test for physical interactions between mER?? and mGluR and will test the role of mGluR on the intracellular effects of E2. In addition to providing important training to the candidate that will help her transition to a career as an independent scientist, the proposed research has the potential to increase the understanding of the mechanisms by which E2 signaling influences food and water intake and has the potential to reveal more general principals of steroid effects on gene expression.