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低温胁迫下大黄鱼Larimichthys crocea中scd1基因的分子克隆与表达分析

Molecular cloning and expression analysis of scd1 gene from large yellow croaker Larimichthys crocea under cold stress.

作者信息

Xu Hao, Zhang Dong Ling, Yu Da Hui, Lv Chang Huan, Luo Hui Yu, Wang Zhi Yong

机构信息

Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture, Fisheries College, Jimei University, Xiamen 361021, PR China.

Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, PR China.

出版信息

Gene. 2015 Aug 15;568(1):100-8. doi: 10.1016/j.gene.2015.05.027. Epub 2015 May 13.

DOI:10.1016/j.gene.2015.05.027
PMID:25979672
Abstract

Desaturation of fatty acids is an important adaptation mechanism to maintain membrane fluidity under cold stress. To comprehend the mechanism of adaptation to low temperatures in fish, we investigated stearoyl-CoA desaturase 1 (SCD1) endocrine expression in the process of cold acclimation from 15°C to 7°C in Larimichthys crocea. The cDNA and genomic sequences of scd1 were cloned and characterized and named as Lcscd1. The cDNA encoded an iron-containing protein of 337 amino acids with functional motifs. The full-length genome sequence of Lcscd1 was composed of 2556 nucleotides, including five exons and four introns. Tissue expression profiles by qPCR and western blot analysis revealed that Lcscd1 was highly expressed in the liver, followed by the brain. The expression of Lcscd1 mRNA in the liver was firstly down-regulated from 15°C to 11°C, and then up-regulated until the first day of 7°C, followed by a decline until the last day. In the brain, the expression showed no significant change from 15°C to 9°C, but then significantly increased until the last day of 7°C. SCD1 protein expression in the liver decreased from 15°C to the first day of 7°C, and then gradually recovered to the starting level. In the brain, SCD1 protein expression maintained rising trends in the whole process. Immunoelectron microscopic analysis showed that SCD1 was localized in fat granules, mitochondria and granular endoplasmic reticulum of hepatic cells, but only in mitochondria of encephalic cells. The results above suggested that SCD1 expression was responsive to both cold and starvation stresses in the liver, but only to cold stress in the brain. In conclusion, these findings suggested that SCD1 may be involved in fish adaptation to cold stress.

摘要

脂肪酸去饱和是在冷应激下维持膜流动性的重要适应机制。为了解鱼类适应低温的机制,我们研究了大黄鱼从15°C冷驯化至7°C过程中硬脂酰辅酶A去饱和酶1(SCD1)的内分泌表达。克隆并鉴定了scd1的cDNA和基因组序列,并将其命名为Lcscd1。该cDNA编码一个含337个氨基酸且具有功能基序的含铁蛋白。Lcscd1的全长基因组序列由2556个核苷酸组成,包括5个外显子和4个内含子。通过qPCR和蛋白质免疫印迹分析的组织表达谱显示,Lcscd1在肝脏中高表达,其次是在脑中。肝脏中Lcscd1 mRNA的表达首先从15°C下调至11°C,然后上调直至7°C的第一天,随后下降直至最后一天。在脑中,从15°C至9°C表达无显著变化,但随后显著增加直至7°C的最后一天。肝脏中SCD1蛋白表达从15°C至7°C的第一天下降,然后逐渐恢复到起始水平。在脑中,SCD1蛋白表达在整个过程中保持上升趋势。免疫电镜分析表明,SCD1定位于肝细胞的脂肪颗粒、线粒体和粗面内质网中,但仅定位于脑细胞的线粒体中。上述结果表明,SCD1的表达在肝脏中对冷应激和饥饿应激均有反应,但在脑中仅对冷应激有反应。总之,这些发现表明SCD1可能参与鱼类对冷应激的适应。

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