Abstract:
Introduction: Elongation of very long-chain fatty acids protein 6 (ELOVL6) played crucial roles in regulating energy expenditure and fatty acid metabolism. Many studies have performed to investigate the physiological roles and regulatory mechanisms of elovl6 in fish and animals, while few studies were reported in crustaceans. Methods: Here we reported on the molecular cloning, tissue distribution and expression profiles in response to dietary fatty acids, ambient salinity and starvation stress in Scylla paramamosain by using rapid amplification of cDNA ends (RACE) and quantitative real-time PCR. Results: Three elovl6 isoforms (named elovl6a, elovl6b and elovl6c) were isolated from S. paramamosain in the present study. The complete sequence of elovl6a was 1345 bp, the full-length sequence of elovl6b was 1419 bp, and the obtained elovl6c sequence was 1375 bp in full length. The elovl6a, elovl6b and elovl6c encoded 287, 329 and 301 amino acids respectively, and exhibited the typical structural features of ELOVL protein family members. Phylogenetic analysis showed that the ELOVL6a from S. paramamosain clustered most closely to ELOVL6 from Portunus trituberculatus and Eriocheir sinensis, while the ELOVL6b and ELOVL6c from S. paramamosain gathered alone into a single branch. Quantitative real-time PCR exhibited that the relatively abundant expression of elovl6b was observed in intestine and stomach, and the elovl6a and elovl6c were highly expressed in hepatopancreas. In addition, studies found that replacing fish oil with soybean oil could significantly increase the transcriptional levels of three elovl6 in hepatopancreas of S. paramamosain, and the expression of elovl6a and elovl6c in hepatopancreas were more sensitive to dietary fatty acids than the elovl6b. Compared with the normal sea water group (27‰), the expression of sterol-regulatory element binding protein1c (srebp-1), elovl6a, elovl6b and elovl6c were upregulated in the low salinity groups, particularly in 7‰. On the contrary, the starvation stress suppressed the expression of srebp-1, elovl6a, elovl6b and elovl6c. Discussion: These results may contribute to understand the functions of elovl6 in fatty acid synthesis and regulatory mechanisms in crustaceans.
摘要:
简介:极长链脂肪酸蛋白6(ELOVL6)的延长在调节能量消耗和脂肪酸代谢中起着至关重要的作用。已经进行了许多研究来研究elovl6在鱼类和动物中的生理作用和调节机制,虽然甲壳类动物的研究很少。方法:我们在这里报道了分子克隆,响应膳食脂肪酸的组织分布和表达谱,通过使用cDNA末端的快速扩增(RACE)和定量实时PCR,在Scyllaparamosain中的环境盐度和饥饿胁迫。结果:三种elovl6亚型(命名为elovl6a,在本研究中,elovl6b和elovl6c)从S.paramamosain分离。elovl6a的完整序列为1345bp,elovl6b的全长序列为1419bp,获得的elovl6c序列全长1375bp。elovl6a,elovl6b和elovl6c分别编码287、329和301个氨基酸,并表现出ELOVL蛋白家族成员的典型结构特征。系统发育分析表明,来自S.paramamosain的ELOVL6a最接近三疣梭子蟹和中华绒螯蟹的ELOVL6,而来自Paramamosain的ELOVL6b和ELOVL6c单独聚集成一个分支。实时定量PCR显示,在肠和胃中观察到elovl6b相对丰富的表达,elovl6a和elovl6c在肝胰腺中高表达。此外,研究发现,用大豆油代替鱼油可以显着提高S.paramamosain肝胰腺中三种elovl6的转录水平,肝胰腺中elovl6a和elovl6c的表达对膳食脂肪酸的敏感性高于elovl6b。与正常海水组(27‰)相比,固醇调节元件结合蛋白1c(srebp-1)的表达,elovl6a,elovl6b和elovl6c在低盐度组中上调,特别是在7‰。相反,饥饿胁迫抑制了srepp-1、elovl6a、elovl6b和elovl6c。讨论:这些结果可能有助于了解elovl6在甲壳类动物脂肪酸合成和调节机制中的功能。