长链多不饱和脂肪酸对三疣梭子蟹卵巢成熟过程中脂肪蓄积影响及其机理的研究

Dietary lipids are considered as an important source of energy and essential fatty acids (EFA) for the crustaceans and long-chain polyunsaturated fatty acids（LC-PUFAs）have also been shown to be of special significance for gonad maturation and brood quality. In this proposed study, we take the swimming crab Portunus trituberculatus as an experimental model. Metabolomics techniques will be utilized to determine the metabolic characteristics in the course of ovary maturation of swimming crabs. Lipoproteins, lipid class and fatty acid profiles will be characterized in the hemolymph, ovary and hepatopancreas, we will conduct to investigate metabolism related genes expression. We will also investigate the effects of different dietary long-chain polyunsaturated fatty acids on Portunus trituberculatus ovarian development and lipid accumulation. We will take advantages of the in vitro organ culture system to study how long-chain polyunsaturated fatty acids affect ovarian lipid absorption and utilization. Understanding the disparity in FA disposal between ovary and lipid accumulation may reveal mechanisms involved in the regulation of long-chain polyunsaturated fatty acids on ovarian development and lipid deposition in Portunus trituberculatus. This pioneering work will pave the road for future research along this line and will provide the basis for understanding Portunus trituberculatus lipid nutrition, gonad maturation and for improving brood quality.

脂类作为壳类动物重要的能量来源和必需脂肪酸，特别是长链多不饱和脂肪酸，参与整个性腺发育过程。本研究以三疣梭子蟹为试验对象，首先在三疣梭子蟹卵巢发育组织学变化分期的基础上，利用代谢组学的技术确定发育各阶段的代谢特征产物，通过测定卵巢发育各期三疣梭子蟹血淋巴中脂蛋白、卵巢和肝胰腺脂类类别和脂肪酸组成以及脂肪积累相关基因表达，研究不同长链不饱和脂肪酸日粮对三疣梭子蟹发育各期卵巢的脂肪蓄积的影响。进而应用体外器官培养模型研究长链脂肪酸与三疣梭子蟹卵巢吸收与利用的关系，从而揭示长链不饱和脂肪酸、卵巢发育与脂肪蓄积三者之间的关系。阐释长链不饱和脂肪酸调控三疣梭子蟹卵巢发育过程中脂肪蓄积的分子机制，完善三疣梭子蟹脂类营养的理论，并为三疣梭子蟹卵巢成熟以及优质亲体培育提供理论依据。

卵巢成熟是一个精心策划的复杂分子过程，需要调控大量基因确保卵母细胞进行适当的发育以促进胚胎发生。卵巢中脂肪酸含量和组成直接影响到甲壳类繁殖，卵子存活和胚胎发育。本项目以我国重要的海水经济蟹类为研究对象，为了更好的阐释长链多不饱和脂肪酸对于三疣梭子蟹卵巢的影响，集成养殖学，代谢组学，分子生物学，细胞生物学，生物信息学和生物化学等多学科手段和方法。运用代谢组学分析脂质类型和组成，通过三疣梭子蟹卵巢发育不同阶段的肝胰腺，卵巢和血淋巴组织中脂质结构和代谢的变化，来阐明三疣梭子蟹的脂质组成差异机理和可能潜在的代谢机制。制作含有不同长链不饱和脂肪酸的配合饲料，通过饲喂三疣梭子蟹，结合体外细胞培养手段，从体内和体外两个方面来阐明长链多不饱和脂肪酸对于三疣梭子蟹卵巢发育的影响，同时辅以组织学手段分析卵巢发育情况和脂肪在卵巢中的蓄积。综合试验结果，DHA可能是影响三疣梭子蟹卵巢发育和脂肪蓄积的最重要长链多不饱和脂肪酸，并提高亲体繁育率和受精卵的质量。项目的开展以及将来在三疣梭子蟹亲体繁殖生物学提供了新的见解和理论基础。本项目已正式发表期刊论文6篇，SCI刊物5篇；已完成项目的各项研究指标。