Somatic regulation of embryonic germline dynamics by isoprenoids

项目概要/摘要 生殖系的发育和保护对下一代的生育力和健康至关重要。 我的长期目标是阐明胚胎生殖细胞的机制，调节和协调， 作为一名独立调查员。新的证据表明，分泌的异戊二烯脂质， 视黄酸和保幼激素在脊椎动物的生殖细胞生命周期中具有不止一种作用， 无脊椎动物，分别。保幼激素信号传导与成人的几个方面有关， 而视黄酸在减数分裂开始时间的调节中具有突出的作用。但没有 尽管这两种类异戊二烯具有高度的结构性， 相似性、交叉激活能力和胚胎生殖系发育过程中共有的表达模式。 基于大量的初步研究，指导这一建议的中心假设是，保幼激素 通过非经典机制影响胚胎生殖细胞动力学， 胚胎生殖过程中的酸性和保幼功能在昆虫和哺乳动物之间是保守的。三 将完成目标，以测试这一假设。首先，保幼激素的影响将与两个确定 接近。时空保幼激素滴度将通过基于光谱的代谢组学测量， 阐明保幼激素的来源。然后，保幼激素对生殖细胞动力学的影响将是 在果蝇胚胎中使用活体成像分析确定。第二，非经典， 转录非依赖性，保幼激素功能的机制将通过蛋白质组学来阐明 鉴定与功能验证偶联的幼鱼结合质膜受体， 下游信号事件将使用D.黑腹胚胎 第三，异戊二烯脂质对胚胎生殖系的保守性将通过阐明 视黄酸对小鼠胚胎生殖细胞动力学的体外和体外研究 充分性分析。与此同时，类异戊二烯迁移后作用的保护将通过 明确了性别特异性差异在D.黑腹生殖系 发展在成功完成这项工作后，对青少年的影响、机制和保护进行了研究。 激素和视黄酸对胚胎生殖系的影响。这些见解将是重要的，因为这些 生物活性类异戊二烯在我们的环境中是普遍存在的。事实上，保幼激素类似物是 全世界使用的许多杀虫剂的成分，并已被证明能激活视黄酸 哺乳动物细胞中的信号传导。因此，了解这些类异戊二烯对胚胎发育的全面影响， 生殖系是至关重要的，努力获得这样的理解，以及配合一般使命的尤尼斯 肯尼迪施赖弗国家儿童健康和人类发展研究所，以确保每个人出生 健康、高产、无病。

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.