Membrane Events Following Hormone Binding to LH Receptor

性状：（由申请人提供）功能性促黄体生成素（LH） 受体对雄性和雌性的生育力都是至关重要的。这些受体 也很重要，因为越来越多的证据表明， LH受体的组成性活性突变导致性早熟 在男性中，与卵巢和睾丸的发育有关 肿瘤的本研究的总体目标是了解 促黄体生成素受体结构在男女性调节中的作用 生育在目的一，我们将确定是否孤立的LH受体形成二聚体 或激素结合后的寡聚体。时间分辨磷光 LH受体旋转扩散和荧光的各向异性测量 将使用单个受体的成像。在目标2中，我们将探讨 LH受体在信号传导中的自相关性。我们将决定 组成型活性LH受体是自相关的， LH受体的交联导致信号传导。在目标3中，我们会问， LH受体在与激素结合后进入脂筏， 受体自身结合必然先于这种易位。筏 分离并跟踪活细胞上的单个LH受体 将被用来确定LH受体是否成为本地化的小 结合激素后的膜微区。在目标4中，我们将确定 膜筏内是否发生LH受体脱敏。隔离 在受体脱敏后的不同时间， 确定是否有LH受体的协调运动， 筏。本实验室研制出了细胞激光光学检测仪 膜的研究，是由少数其他实验室等同。的应用 这些仪器方法将提供深入了解膜事件 伴随LH受体介导的信号转导和受体 脱敏这些研究还将借鉴PI的经验， 以及科林·克莱博士的专业知识， 和生物技术实验室的神经生物学家迈克尔·塔姆昆博士， 科罗拉多州立大学。

DESCRIPTION: (provided by applicant) Functional luteinizing hormone (LH) receptors are critical to fertility in both males and females. These receptors are also important because of increasing evidence that naturally-occurring, constitutively active mutations of the LH receptor lead to precocious puberty in males and are implicated in the development of ovarian and testicular tumors. The overall goal of this research is to understand the role of luteinizing hormone receptor structure in regulation of male and female fertility. In Aim I we will determine whether isolated LH receptors form dimers or oligomers following hormone binding. Time-resolved phosphorescence anisotropy measurements of LH receptor rotational diffusion and fluorescence imaging of single receptors will be used. In Aim 2 we will explore the role of the LH receptor self-association in signaling. We will determine whether constitutively active LH receptors are self-associated and whether extensive crosslinking of LH receptors results in signaling. In Aim 3 we will ask whether LH receptors move into lipid rafts following binding of hormone and whether receptor self-association necessarily precedes this translocation. Raft isolation together with tracking of individual LH receptors on viable cells will be used to determine whether LH receptors become localized in small membrane microdomains following binding of hormone. In Aim 4 we will determine whether LH receptor desensitization occurs within membrane rafts. The isolation of membrane rafts at various times following receptor desensitization will establish whether there is orchestrated movement of LH receptors into and out of rafts. Our laboratory has developed laser optical instrumentation for cell membrane studies that are equaled by few other laboratories. The application of these instrumental methods will provide insight into membrane events accompanying LH-receptor mediated signal transduction and receptor desensitization. These studies will also draw on the experience of the PI as well as the expertise of Dr. Colin Clay, a member of the Animal Reproduction and Biotechnology Laboratory, and of Dr. Michael Tamkun, a neurobiologist at Colorado State University.