Lipid Mediators in Drosophila Spermatogenesis

项目摘要 细胞通过激活下游信号的分泌分子相互交流 途径。许多研究充分的途径依赖于蛋白质作为细胞之间的信使，但某些脂质 也可以作为沟通的媒介。脂质信号分子以膜的潜伏形式储存 磷脂，其被磷脂酶A2酶分解成生物活性的溶血磷脂和脂肪磷脂。 acids.溶血磷脂酰基转移酶催化称为溶血磷脂酰基转移酶的生物化学途径中的逆反应。 土地循环Lands循环及其调节的脂质介质在哺乳动物血管中发挥作用， 神经，免疫和生殖系统，但哺乳动物研究的解释已经复杂化， 多效性、冗余和技术限制。黑腹果蝇是一个首要的模型系统， 研究细胞间的通讯，因为它在进化上是保守的，但简单，世代短 与哺乳动物相比。以前对果蝇的研究几乎完全集中在蛋白质信号上。 这里提出的研究将利用成熟的遗传学方法， 这些工具包括敲除突变体、组织特异性RNAi、基因过表达和错误表达。 果蝇Lands循环酰基转移酶Oys和内斯是精子发生所必需的。具体目标1将 分析Oys和内斯在果蝇睾丸中的作用机制，特别是研究它们是如何在果蝇睾丸中发挥作用的。 它们介导体细胞支持细胞和生殖细胞之间的通讯。与其他基因相互作用 将测定影响溶血磷脂和脂肪酸水平的途径。质谱分析将是 用于评估oys内斯突变体中脂质水平的变化。具体目标2将调查土地周期 磷脂酶A2和假定的溶血磷脂酶D在睾丸中，再次使用遗传学和 质谱分析法来在补充方法中，睾丸将与化学酶抑制剂一起培养。 具体目标3使用睾丸系统筛选土地循环的显性遗传修饰剂，以 鉴定该途径的调节因子和信号应答因子。研究脂质介导的 果蝇模型中的信号传导将阐明许多基本的和保守的范式，正如 蛋白质信号的情况下，并将有助于阐明生物活性脂质在疾病中的作用。异常脂质信号传导 与代谢综合征、动脉粥样硬化、自身免疫、哮喘 神经系统疾病、癌症和不育症。此外，脂肪酸摄入的饮食调节具有重要意义。 对健康、生殖和发育的影响，但其机制尚不清楚。这 研究将与本科生和学士后的学生只进行，对本科 叶史瓦大学校园，从而有力地解决了R15区域计划的指令： 增强本科生的科研经验，强化科研环境， 大学校园，并让大学生接触发育模式生物。

PROJECT SUMMARY Cells communicate with each other via secreted molecules that activate downstream signaling pathways. Many well-studied pathways rely on proteins to act as messengers between cells, but certain lipids also can act as communication mediators. Lipid signaling molecules are stored in latent form as membrane phospholipids, which are broken down by phospholipase A2 enzymes into bioactive lysophospholipids and fatty acids. Lysophospholipid acyltransferases catalyze the reverse reaction in a biochemical pathway known as the Lands Cycle. The Lands Cycle and the lipid mediators it regulates play roles in the mammalian vascular, nervous, immune, and reproductive systems, but interpretation of mammalian studies has been complicated by pleiotropy, redundancy, and technical limitations. Drosophila melanogaster is a premier model system for studying intercellular communication due to its evolutionary conservation but simplicity and short generation time compared to mammals. Prior studies in Drosophila have focused almost exclusively on protein signaling. The research proposed here will investigate lipid signaling in the fly model, using the well-developed genetic tools available, including knockout mutants, tissue specific RNAi, gene overexpression and misexpression. Drosophila Lands Cycle acyltransferases Oys and Nes are required for spermatogenesis. Specific Aim 1 will analyze the mechanism by which Oys and Nes function in the Drosophila testis, specifically examining how they mediate communication between somatic support cells and the germline. Genetic interactions with other pathways that affect levels of lysophospholipids and fatty acids will be assayed. Mass spectrometry will be used to assess changes in lipid levels in oys nes mutants. Specific Aim 2 will investigate Lands Cycle phospholipases A2 and a putative lysophospholipase D in the testis, again using a combination of genetics and mass spectrometry. In a complementary approach, testes will be cultured with chemical enzyme inhibitors. Specific Aim 3 uses the testis system to screen for dominant genetic modifiers of the Lands Cycle, in order to identify regulators and signal response factors of this pathway. It is expected that studying lipid mediated signaling in the Drosophila model will illuminate many fundamental and conserved paradigms, as has been the case for protein signaling, and will help elucidate the roles of bioactive lipids in disease. Aberrant lipid signaling has been implicated in such diverse disorders as metabolic syndrome, atherosclerosis, autoimmunity, asthma, neurological disease, cancer, and infertility. Furthermore, dietary modulation of fatty acid intake has important consequences for health, reproduction, and development, but the mechanisms are not well understood. This research will be conducted with undergraduate and post-baccalaureate students only, on the undergraduate campus of Yeshiva University, thereby strongly addressing the directives of the R15 AREA program: to enhance the research experience of undergraduates, to strengthen the research environment on undergraduate campuses, and to expose undergraduates to developmental model organisms.