Fatty acid metabolism and signaling during zebrafish development

斑马鱼发育过程中的脂肪酸代谢和信号传导

• 批准号: 7409459
• 负责人: Rosa Linda Miyares
• 金额: $ 3.01万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-15 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7409459
• 关键词: Acyl Coenzyme A; Address; Attenuated; Blood Cells; Blood Circulation; Cells; Cellular Morphology; Clinical; Coenzyme A Ligases; Coupled; Data; Defect; Development; Developmental Process; Embryo; Environment; Enzymes; Erythrocytes; Erythroid; Erythroid Cells; Erythropoiesis; Fatty Acids; Fishes; Genes; Goals; Hematological Disease; Human; Image; Knowledge; Larva; Lipids; Membrane; Messenger RNA; Metabolism; Microscopy; Monitor; Nature; Orthologous Gene; Pathway interactions; Pattern; Phenotype; Physiology; Play; Proteins; Reagent; Role; Rosa; Signal Transduction; Site; Staining method; Stains; Techniques; Testing; Time; Tissues; Transcript; Transgenic Organisms; Transport Process; Very Long Chain Fatty Acid; Yolk Cell; Zebrafish; absorption; fatty acid metabolism; fatty acid-transport protein; in vivo; leukemia; lipid metabolism; liquid chromatography mass spectroscopy; long chain fatty acid; man; migration; research study

DESCRIPTION (provided by applicant): Lipid metabolism is essential for all cells and most likely every developmental process, and yet many questions remain regarding mechanisms of lipid processing and transport. The Farber lab uses the developing zebrafish to study the roles of long-chain acyl-CoA synthetases (Acsls) and fatty acid transport proteins (Fatps), enzymes that process and transport long-chain and very long-chain fatty acids. My long term goal is to understand the role of one of these proteins, fatty acid transport protein 2a (Fatp2a), during zebrafish development and ultimately in human physiology. Fatp2a is an ortholog of human Fatp2, an enzyme that both transports long-chain fatty acids across membranes and activates very long-chain fatty acids into fatty acyl-CoAs. My preliminary studies demonstrate that loss of Fatp2a in zebrafish larvae profoundly attenuates primitive red blood cell development (erythropoiesis). Interestingly, fatp2a transcript is primarily expressed not in the site of primitive erythropoiesis, but in the yolk syncytial layer (YSL). Because the YSL surrounds the yolk cell and expresses many genes important for lipid absorption, metabolism, and packaging (Marza et al 2005; Farber et al, unpublished), we believe the YSL acts to absorb lipids from the yolk and package them for delivery to the developing embryo. The proposed studies will test the hypothesis that Fatp2a activates or transports a specific fatty acid from the yolk that is necessary for proper primitive erythropoiesis. The following specific aims will help elucidate the nature of the phenotype and reveal the substrate that Fatp2a activates and/or imports to influence erythropoiesis. I will explore the role of Fatp2a in proliferation, migration and maturation of erythroid cells by using different staining techniques and in vivo time-lapse microscopy. I will explore the requirement for Fatp2a in a cell autonomous or non-autonomous manner by microinjecting antisense reagents and mRNA in a tissue specific manner as well as creating transgenic fish lines. Finally I will reveal Fatp2a's substrate using mass liquid chromatography/mass spectroscopy. The preliminary data, coupled with the proposed experiments will address a new and unexpected role for Fatp2 activity in regulating blood cell development. These results will not only increase our knowledge of this important developmental pathway, but may have clinical implications for the treatment of devastating blood diseases like some leukemias.

描述（由申请人提供）：脂质代谢对所有细胞和几乎所有发育过程都是必不可少的，然而关于脂质加工和运输的机制仍然存在许多问题。法伯实验室利用发育中的斑马鱼来研究长链酰基辅酶a合成酶（Acsls）和脂肪酸转运蛋白（Fatps）的作用，这些酶处理和转运长链和超长链脂肪酸。我的长期目标是了解其中一种蛋白质脂肪酸转运蛋白2a （Fatp2a）在斑马鱼发育过程中的作用，并最终在人体生理学中发挥作用。Fatp2a是人类Fatp2的同源物，Fatp2是一种酶，既可以将长链脂肪酸跨膜运输，也可以将非常长的链脂肪酸激活为脂肪酰基辅酶a。我的初步研究表明，斑马鱼幼体中Fatp2a的缺失会严重削弱原始红细胞的发育（红细胞生成）。有趣的是，fatp2a转录本主要不是在原始红细胞生成位点表达，而是在卵黄合胞层（YSL）表达。由于YSL包围着卵黄细胞并表达许多对脂质吸收、代谢和包装重要的基因（Marza et al, 2005; Farber et al，未发表），我们认为YSL从卵黄中吸收脂质并将其包装以传递给发育中的胚胎。拟议的研究将验证Fatp2a激活或从蛋黄中运输一种特定的脂肪酸的假设，这种脂肪酸是正常的原始红细胞生成所必需的。以下具体目的将有助于阐明表型的性质，并揭示Fatp2a激活和/或输入影响红细胞生成的底物。我将通过使用不同的染色技术和体内延时显微镜来探索Fatp2a在红细胞增殖、迁移和成熟中的作用。我将通过以组织特异性的方式微注射反义试剂和mRNA以及创建转基因鱼种来探索对细胞自主或非自主方式的Fatp2a的要求。最后，我将使用质液相色谱/质谱法揭示Fatp2a的底物。这些初步数据，加上提出的实验，将解决Fatp2活性在调节血细胞发育中的一个新的和意想不到的作用。这些结果不仅将增加我们对这一重要发育途径的认识，而且可能对治疗一些破坏性血液疾病（如白血病）具有临床意义。