Somatic regulation of embryonic germline dynamics by isoprenoids

项目摘要/摘要 生殖系的发展和保护对下一代的生育和健康至关重要。 我的长期目标是阐明胚胎生殖系的机制、调节和协调。 以独立调查员的身份进行调查。新出现的证据表明，分泌的异戊二烯类脂， 维甲酸和保幼激素，在脊椎动物和 分别是无脊椎动物。保幼激素信号在成人的几个方面都有牵连 配子发生，而维甲酸在减数分裂开始时间的调节中起着重要作用。然而，没有 这两个异戊二烯类化合物的共同功能迄今已被发现，尽管高度的结构 胚胎生殖系发育过程中的相似性、交叉激活能力和共同的表达模式。 基于大量的初步研究，指导这一提议的中心假设是保幼激素 通过非经典机制和维甲酸两种功能影响胚胎生殖细胞动力学 昆虫和哺乳动物在胚胎发育过程中的酸性和幼体功能是保守的。三 将完成AIMS来检验这一假说。首先，保幼激素的影响将通过两个方面来确定 接近了。基于光谱的代谢组学将测量时空保幼激素效价 阐明保幼激素的来源。那么，保幼激素对生殖细胞动力学的影响将是 通过对果蝇胚胎进行活体成像分析来确定。第二，非古典的， 蛋白质组学将阐明保幼激素作用的转录非依赖性机制 结合功能验证的保幼激素结合质膜受体的鉴定， 下游的信号事件将使用活的成像显微镜在黑腹果蝇胚胎中阐明。 第三，异戊二烯类脂类在胚胎生殖系上的保守性将通过阐明 维甲酸对小鼠胚胎生殖细胞动力学的影响 充分性分析。与此同时，类异戊二烯迁移后作用的保守性将通过以下方式进行审查 确定性别差异对黑腹果蝇种质中保幼激素效价的影响 发展。这项工作圆满完成后，对青少年的影响、机制和保护 胚胎生殖系上的激素和维甲酸将被定义。这些见解将非常重要，因为 生物活性异戊二烯类化合物普遍存在于我们的环境中。事实上，保幼激素类似物是活性的 世界各地使用的许多杀虫剂的成分，已被证明能激活维甲酸 哺乳动物细胞中的信号。因此，了解这些异戊二烯类化合物对胚胎的全面影响 生殖系是至关重要的，为获得这种理解所作的努力很好地符合尤尼斯的总体使命 肯尼迪·施莱弗国家儿童健康和人类发展研究所，确保每个人出生 健康，多产，没有疾病。

PROJECT SUMMARY/ABSTRACT The development and protection of the germline is critical to the fertility and the health of the next generation. My long-term objective is to elucidate the mechanisms, regulation and coordination of embryonic germline processes as an independent investigator. Emerging evidence suggests that the secreted isoprene lipids, retinoic acids and juvenile hormones, have more than one role in the germline lifecycle in vertebrate and invertebrate animals, respectively. Juvenile hormone signaling has been implicated in several aspects adult gametogenesis while retinoic acid has a prominent role in the regulation of meiotic onset timing. However, no shared function for these two isoprenoids have been found to-date despite a high degree of structural similarity, cross-activation capacity, and shared expression patterns during embryonic germline development. Based on a wealth of preliminary studies, the central hypothesis guiding this proposal is that juvenile hormone impacts embryonic germ cell dynamics through a non-classical mechanism and that two functions of retinoic acid and juvenile function in embryonic germ processes are conserved between insects and mammals. Three aims will be completed to test this hypothesis. First, the impact of juvenile hormone will be determined with two approaches. Spatiotemporal juvenile hormone titers will be measured spectrometry-based metabolomics to elucidate the source of juvenile hormone. Then, the impact of juvenile hormone on germ cell dynamics will be determined using live imaging analysis in Drosophila melanogaster embryos. Second, the non-classical, transcription-independent, mechanism of juvenile hormone function will be elucidated by proteomic identification of the juvenile hormone-binding plasma membrane receptors coupled to functional verification, Downstream signaling events will be elucidated using live imaging microscopy in D. melanogaster embryos. Third, the conservation of isoprene lipids on the embryonic germline will be defined by elucidating the impact of retinoic acids on germ cell dynamics in Mus musculus embryos by ex vivo and in vitro necessity and sufficiency assays. Concomitantly, conservation of the post-migratory role for isoprenoids will be examined by defining the role of sex-specific differences in juvenile hormone titers in D. melanogaster germline development. Upon successful completion of this work, the impact, mechanism and conservation of juvenile hormone and retinoic acid on the embryonic germline will be defined. These insights will be important as these bioactive isoprenoids are pervasive in our environment. In fact, juvenile hormone analogues are the active ingredient of many insecticides used throughout the world and have been shown to activate retinoic acid signaling in mammalian cells. Thus, understanding the full impact of these isoprenoids on the embryonic germline is critical and the efforts to gain such understanding align well the general mission of the Eunice Kennedy Shriver National Institute of Child Health and Human Development to ensure every person is born healthy, productive, are free of disease.