Molecular mechanisms of steroid hormone secretion and trafficking

项目摘要/摘要 类固醇激素是一大类分子，在儿童发育过程中发挥着关键作用。在.期间 在人类青春期，性腺类固醇激素分泌增加会产生第二性征 例如乳房发育和面部毛发的出现。因为类固醇的重要作用 成熟过程中的激素，童年时期类固醇激素信号的中断可能会导致 持续到成年期的发育缺陷。了解生物多样性的机制和调控机制 因此，在正常和病理条件下，类固醇激素信号在很大程度上有助于 促进儿童健康发展。这个项目的最终目标是阐明目前尚不清楚的 果蝇类固醇激素释放和运输的机制和调控机制 作为一种模式生物。为了实现这一目的，PI将检验昆虫类固醇 激素蜕皮激素以囊泡介导的方式从类固醇合成组织中分泌出来，挑战 认为所有类固醇激素都是通过自由扩散分泌的传统观点。在项目的第一个指导阶段 在这个项目中，PI将与他在明尼苏达大学的导师迈克尔·奥康纳密切合作，开发出 一些关键的体外方法是阐明他的假说所必需的。这些方法包括 蜕皮激素免疫组织化学检测、体外转运蛋白测定和体外类固醇合成组织培养。 这个拟议项目的第一步将帮助PI掌握各种生物化学和细胞生物学技术 需要进行项目的下一步工作。在指导阶段，PI还将经历广泛的 关于教学和科学交流的培训，这将有助于HIS的下一个独立阶段 职业生涯。在随后的独立调查员阶段，独立调查员将致力于 通过筛选起作用的G蛋白偶联受体，推测囊泡介导的蜕皮激素释放 类固醇生成组织。他还将筛选外周摄取蜕皮激素所需的可能的蜕皮激素进口者 纸巾。这些方法应该告诉我们，这种新的类固醇激素分泌机制和 贩运在不同的生物体之间是保守的。从长远来看，PI的工作有可能改变 类固醇激素作用的范例，并将影响对发育和疾病的广泛研究 流程。

Project Summary/Abstract Steroid hormones are a large family of molecules that play pivotal roles during childhood development. During puberty in humans, elevated secretion of gonadal steroid hormones produces secondary sex characteristics such as breast development and the appearance of facial hair. Because of such important roles of steroid hormones in maturation processes, disruption of steroid hormone signaling during childhood can cause developmental defects that last into adulthood. Understanding the machinery and regulatory mechanisms of steroid hormone signaling in normal as well as pathological conditions, therefore, contributes greatly to the promotion of healthy childhood development. The ultimate goal of this project is to elucidate as-yet-unknown machinery and regulatory mechanisms of steroid hormone release and trafficking, by using the fruitfly Drosophila as a model organism. In order to accomplish this purpose, the PI will test the hypothesis that the insect steroid hormone ecdysone is secreted from the steroidogenic tissue in a vesicle-mediated manner, challenging the conventional idea that all steroid hormones are secreted by free diffusion. During the first mentored phase of the project, the PI will work closely with his mentor, Michael O'Connor, at the University of Minnesota to develop some key in vitro methods necessary to elucidate his hypothesis. Those methods include the immunohistochemical detection of ecdysone, in vitro transporter assay and in vitro steroidogenic tissue culture. This initial step of the proposed project will help the PI master various biochemistry and cell biology techniques required to conduct the next step of the project. During the mentored phase, the PI will also undergo extensive training on teaching and scientific communication, which will be helpful in the next independent phase of his career. In the subsequent independent investigator phase, the PI will work on the regulatory mechanisms of the putative vesicle-mediated ecdysone release, by screening G protein-coupled receptors working in the steroidogenic tissue. He will also screen for a putative ecdysone importer required for its uptake by peripheral tissues. These approaches should tell us how well this novel machinery of steroid hormone secretion and trafficking is conserved among different organisms. In the long run, the PI's work has the potential to shift the paradigm of steroid hormone action and will impact a vast range of research on developmental and disease processes.