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杨露露, 赵伟芳, 徐歆歆, 张玉茹, 曹香林, 聂国兴, 卢荣华. 草鱼Isthmin-1基因序列分析及转录水平的营养调控[J]. 水生生物学报, 2023, 47(8): 1278-1283. DOI: 10.7541/2023.2022.0195
引用本文: 杨露露, 赵伟芳, 徐歆歆, 张玉茹, 曹香林, 聂国兴, 卢荣华. 草鱼Isthmin-1基因序列分析及转录水平的营养调控[J]. 水生生物学报, 2023, 47(8): 1278-1283. DOI: 10.7541/2023.2022.0195
YANG Lu-Lu, ZHAO Wei-Fang, XU Xin-Xin, ZHANG Yu-Ru, CAO Xiang-Lin, NIE Guo-Xing, LU Rong-Hua. SEQUENCE ANALYSIS AND NUTRITIONAL REGULATION OF ISTHMIN-1 IN GRASS CARP (CTENOPHARYNGODON IDELLA)[J]. ACTA HYDROBIOLOGICA SINICA, 2023, 47(8): 1278-1283. DOI: 10.7541/2023.2022.0195
Citation: YANG Lu-Lu, ZHAO Wei-Fang, XU Xin-Xin, ZHANG Yu-Ru, CAO Xiang-Lin, NIE Guo-Xing, LU Rong-Hua. SEQUENCE ANALYSIS AND NUTRITIONAL REGULATION OF ISTHMIN-1 IN GRASS CARP (CTENOPHARYNGODON IDELLA)[J]. ACTA HYDROBIOLOGICA SINICA, 2023, 47(8): 1278-1283. DOI: 10.7541/2023.2022.0195

草鱼Isthmin-1基因序列分析及转录水平的营养调控

SEQUENCE ANALYSIS AND NUTRITIONAL REGULATION OF ISTHMIN-1 IN GRASS CARP (CTENOPHARYNGODON IDELLA)

  • 摘要: 为探讨Isthmin-1(Ism-1)在草鱼糖和脂类代谢中的作用, 研究采用RT-PCR技术克隆草鱼Ism-1的开放阅读框(ORF), 生物信息学分析Ism-1及其编码的氨基酸序列, RT-qPCR技术检测Ism-1在草鱼各组织中的分布特点, 并在细胞和活体水平上分析不同营养条件下Ism-1 mRNA的表达变化。结果显示, 成功克隆草鱼Ism-1的ORF区。序列分析表明, 草鱼Ism-1基因开放读码框为1380 bp, 编码459个氨基酸, 预测该蛋白相对分子量为50.96 kD。氨基酸多序列比对和系统进化树分析显示, 草鱼Ism-1与黑头软口鲦(Pimephales promelas)的进化关系最近(氨基酸相似度为96.51%)。Ism-1在草鱼各组织中均有表达, 在红肌中表达量最高, 其次是鳃、脑和白肌等组织。饥饿再投喂实验结果表明, 饥饿14d后肝胰脏中Ism-1的表达量显著上调(P<0.05), 恢复投喂后表达量降低, 但仍显著高于对照组(P<0.05), 白肌中Ism-1的表达量在饥饿和再投喂后均显著增加(P<0.05)。腹腔注射不同浓度的胰高血糖素显著下调草鱼肝胰脏中Ism-1的mRNA水平(P<0.05), 油酸孵育草鱼原代肝细胞24h时, Ism-1的表达量显著减少(P<0.001)。结果提示, 草鱼Ism-1可能参与机体糖和脂类的代谢过程调控, 这为后续研究Ism-1的功能提供了基础数据。

     

    Abstract: In order to explore the role of Isthmin-1 (Ism-1) in glucose and lipid metabolism of grass carp, we cloned the open reading frame (ORF) of Ism-1 by RT-PCR, and the sequence was analyzed by bioinformatics technology. The distribution characteristics of Ism-1 in different tissues of grass carp were detected by RT-qPCR, and the expression of Ism-1 under different nutritional conditions were analyzed in vitro and in vivo. In this study, ORF region of Ism-1 was successfully cloned. Sequence analysis showed that the ORF region of Ism-1 was 1380 bp, encoding 459 amino acids, and the relative molecular weight of protein was 50.96 kD. Amino acid multiple sequence alignment and phylogenetic tree analysis showed that the evolutionary relationship of Ism-1 between grass carp and fathead minnow was the closest (amino acid similarity was up to 96.51%). Ism-1 was widely expressed in multiple tissues, and had higher expression in red muscle, next were gill, brain and white muscle of grass carp. The fasting and refeeding experiment indicated the expression of Ism-1 in hepatopancreas was significantly upregulated after 14 days of starvation treatment (P<0.05), and the expression decreased after refeeding, but was still significantly higher than that of control group (P<0.05). The expression ofIsm-1 in white muscle was significantly up-regulated after starvation and refeeding (P<0.05). The results of intraperitoneal injection of glucagon experiment eventuated that the expression ofIsm-1 in hepatopancreas was significantly downregulated compared with control group (P<0.05). In addition, the expression ofIsm-1 was significantly decreased (P<0.001) in hepatocytes after treatment with 80 μg/mL oleic acid for 24h. Overall, this research confirmedIsm-1 may be involved in the regulation of glucose and lipid metabolism of grass carp, which would provide essential data for the following study of the function of Ism-1.

     

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