Yang Zhigang, Guo Zihao, Ji Lianyuan, Zeng Qitao, Wang Yao, Yang Xiaozhen, Cheng Yongxu
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Science, Shanghai Ocean University, Huchenghuan Road 999, Lingang New District, Shanghai 201306, PR China.
Comp Biochem Physiol B Biochem Mol Biol. 2013 Jun;165(2):99-105. doi: 10.1016/j.cbpb.2013.03.010. Epub 2013 Mar 16.
Fatty acyl Δ6-desaturase is the rate-limiting enzyme in the biosynthetic pathway of highly unsaturated fatty acids (HUFAs) in vertebrates. In this report, a fatty acyl Δ6-desaturase-like cDNA was cloned from the hepatopancreas of Eriocheir sinensis (Chinese mitten crab) and characterized by performing rapid-amplification of cDNA ends. The 2278-bp long full-length cDNA encodes a polypeptide with 442 amino acids. Gene expression analysis via real-time quantitative polymerase chain reaction revealed that the fatty acyl Δ6-desaturase-like transcripts are widely distributed in various tissues, with high expression levels in the hepatopancreas and cranial ganglia. This study focuses on the nutritional regulation of genes involved in the HUFA biosynthetic pathway in Chinese mitten crab. A feeding trial was performed whereby crablets were fed for 238 d with four different diets: control diet without oil lipids (added with 3% basic lipid of the fundamental diets); fish oil diet (FO; added with 3% of the fundamental diets); soybean oil diet (SO; added with 3% of the fundamental diets); and FO/SO diet (1:1; added with 3% of the fundamental diets). The hepatopancreas of crabs sampled at 168 d and 238 d to determine the effects on fatty acyl Δ6-desaturase-like mRNA expression. The results show that the expression of fatty acyl Δ6-desaturase-like is higher in the hepatopancreas of crabs fed with SO diet than those fed with FO diet. Furthermore, gene expression increased by 2.45-fold in the hepatopancreas of crabs fed with SO after 238 d than those fed after 168 d but remained steady for those fed with FO after 238 d.
脂肪酰Δ6-去饱和酶是脊椎动物中高度不饱和脂肪酸(HUFA)生物合成途径中的限速酶。在本报告中,从中华绒螯蟹(河蟹)的肝胰腺中克隆了一个类脂肪酰Δ6-去饱和酶cDNA,并通过cDNA末端快速扩增进行了表征。这个2278bp长的全长cDNA编码一个含有442个氨基酸的多肽。通过实时定量聚合酶链反应进行的基因表达分析表明,类脂肪酰Δ6-去饱和酶转录本广泛分布于各种组织中,在肝胰腺和脑神经节中表达水平较高。本研究聚焦于河蟹中参与HUFA生物合成途径的基因的营养调控。进行了一项投喂试验,用四种不同的饲料喂养蟹苗238天:不含油脂的对照饲料(添加3%基础饲料的基础脂质);鱼油饲料(FO;添加3%基础饲料);大豆油饲料(SO;添加3%基础饲料);以及FO/SO饲料(1:1;添加3%基础饲料)。在第168天和第238天对蟹进行取样,取其肝胰腺以确定对类脂肪酰Δ6-去饱和酶mRNA表达的影响。结果表明,喂食SO饲料的蟹的肝胰腺中类脂肪酰Δ6-去饱和酶的表达高于喂食FO饲料的蟹。此外,喂食SO饲料的蟹在238天后肝胰腺中的基因表达比168天后增加了2.45倍,但喂食FO饲料的蟹在238天后保持稳定。