Estrogen receptor activation in the CNS

描述(申请人提供)：大脑雌激素受体(ER)，就像外周ER一样，可以被雌激素直接激活，也可能通过非配体激活间接激活。配体非依赖通路可能介导体液因素和来自环境的传入输入对内质网神经元的影响。这项试探性/开发性拨款申请的目的是评估并进一步开发一种新型的转基因小鼠品系--ER动作指示器(ERIN)品系(它具有转基因，在与β-半乳糖苷酶报告基因相连的启动子上包含雌激素反应元件)，用于研究大脑中ER的非配体激活。首先，将使用免疫细胞化学技术在细胞群体中表征神经元对雌二醇的转基因反应，这是必不可少的第一步。然后将评估该小鼠品系对研究配体非依赖性激活的有用性。这项研究的长期目标是了解由神经递质和其他体液因素介导的来自环境的传入输入作用于神经元内质网以调节神经生理和神经内分泌事件的细胞过程。在一些实验中，ERIN小鼠与带有ERAlpha或ERbeta基因中断的小鼠品系杂交，将被用来评估每个ER亚型在对激素、神经递质和第二信使途径的转基因反应中的作用。这项申请是作为R21提交的，因为它的目的是探索使用一种高度创新的方法(Erin小鼠)来研究一个新的领域，即大脑，并为这一领域的额外工作开发这一新技术，并开发一系列数据，以此为基础进行重大的未来研究。这项初步研究对于未来研究男性和女性大脑中ER的配体独立激活的分子后果至关重要，然后可以以前所未有的方式进行研究。

DESCRIPTION (provided by applicant): Brain estrogen receptors (ERs), like peripheral ERs, can be activated directly by estrogens, and presumably indirectly by ligand independent activation. The ligand independent pathway may mediate the effects of humoral factors and of afferent input derived from the environment on ER-containing neurons. The purpose of this Exploratory/Developmental grant application is to assess the usefulness of, and then further develop, a novel, transgenic, mouse strain, the ER action indicator (ERIN) strain of mice (which has a transgene, containing estrogen response elements on a promoter linked to a beta-galactosidase reporter gene) for studies of ligand independent activation of ERs in the brain. First the neuronal transgene response to estradiol will be characterized in populations of cells using an immunocytochemical technique, an essential first step. Then the usefulness of this mouse strain for studies of ligand-independent activation will be assessed. The long-range goal of this research is to understand the cellular processes by which afferent input from the environment, mediated by neurotransmitters and other humoral factors, act on neuronal ERs to modulate neuronal physiology and neuroendocrine events. In some experiments, ERIN mice crossed with strains of mice with ERalpha- or ERbeta-gene disruptions will be used to assess the role of each ER subtype in transgene responses to hormones, neurotransmitters, and second messenger pathways. This application is being submitted as an R21, because its purpose is to explore the use of a highly innovative approach (the ERIN mice) to a new area, the brain, and to develop this novel technology for additional work in this area, and to develop a body of data upon which significant future research may be built. This initial research is essential for future work on the molecular consequences of ligand independent activation of ERs in the brain of males and females, which can then be studied in an unprecedented way.