Somatic regulation of embryonic germline dynamics by isoprenoids

类异戊二烯对胚胎种系动力学的体细胞调节

基本信息

批准号：
10682515
负责人：
Lacy J Barton
金额：
$ 24.12万
依托单位：
UNIVERSITY OF TEXAS SAN ANTONIO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10682515
关键词：
Adolescent Adult Animals Binding Biogenesis Biological Assay Cell membrane Cell physiology Coupled Data Development Disease Drosophila genus Drosophila melanogaster Embryo Embryonic Development Ensure Environment Enzymes Event Exhibits Fertility Foundations G-Protein-Coupled Receptors Gametogenesis Genes Genetic Suppression Genetic Transcription Germ Germ Cells Gonadal structure Health Hormone secretion House mice Human Image In Vitro Insecta Insecticides Institution Invertebrates Isoprene Juvenile Hormones Life Cycle Stages Lipids Mammalian Cell Mammals Mass Spectrum Analysis Measures Mediating Meiosis Mentors Mission Modeling Mus National Institute of Child Health and Human Development Nature Oogenesis Pattern Persons Pharmaceutical Preparations Phospholipase C Positioning Attribute Process Productivity Proteins Proteomics Receptor Activation Regulation Research Research Personnel Role Sex Differences Signal Transduction Source Spectrometry Spermatogenesis Structure of primordial sex cell Testing Time Training Tretinoin Work animal imaging cell motility experimental study hormonal signals hormone analog in vitro Assay in vivo insight isoprenoid male metabolomics microscopic imaging migration next generation non-genomic novel prevent programs receptor sex sex determination spatiotemporal

项目摘要

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.

项目概要/摘要种系的发育和保护对于下一代的生育能力和健康至关重要。我的长期目标是阐明胚胎种系的机制、调控和协调作为独立调查员的流程。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种系生命周期中具有不止一种作用分别是无脊椎动物。保幼激素信号传导与成人的几个方面有关配子发生，而视黄酸在调节减数分裂开始时间方面具有突出作用。然而，没有迄今为止，尽管结构高度相似，但已发现这两种类异戊二烯具有共同的功能。胚胎种系发育过程中的相似性、交叉激活能力和共享表达模式。基于大量的初步研究，指导该提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，并且视黄酸的两个功能胚胎生殖过程中的酸和幼体功能在昆虫和哺乳动物之间是保守的。三将完成测试这一假设的目标。首先，保幼激素的影响将通过两个因素来确定接近。时空保幼激素滴度将通过基于光谱分析的代谢组学进行测量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使用果蝇胚胎的实时成像分析确定。二、非古典、不依赖转录的保幼激素功能机制将通过蛋白质组学来阐明保幼激素结合质膜受体的鉴定以及功能验证，将使用黑腹果蝇胚胎中的实时成像显微镜来阐明下游信号传导事件。第三，异戊二烯脂质对胚胎种系的保护将通过阐明以下因素的影响来定义：视黄酸对小家鼠胚胎生殖细胞动力学的离体和体外必要性和充分性分析。与此同时，类异戊二烯迁移后作用的保护将由确定黑腹果蝇种系中保幼激素滴度的性别特异性差异的作用发展。这项工作顺利完成后，对青少年的影响、机制和保护将定义胚胎种系上的激素和视黄酸。这些见解将很重要，因为具有生物活性的类异戊二烯在我们的环境中普遍存在。事实上，保幼激素类似物是活性物质世界各地使用的许多杀虫剂的成分，已被证明可以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的全面影响种系至关重要，获得这种理解的努力与尤尼斯的总体使命非常吻合肯尼迪施赖弗国家儿童健康和人类发展研究所确保每个人的出生健康、富有成效、无疾病。