西伯利亚鲟AdipoR克隆、组织分布及其对禁食的响应

CLONING AND TISSUE DISTRIBUTION OF ADIPONECTIN RECEPTOR AND IT’S RESPONSE ON FASTING/REFEEDING IN SIBERIAN STURGEON (ACIPENSER BAERI)

  • 摘要: 为了探究西伯利亚鲟脂联素受体的表达特征及其对能量状态变化的响应, 研究采用RACE方法克隆获得AdipoR1和AdipoR2的cDNA全长。西伯利亚鲟AdipoR1 cDNA全长为2013 bp, 开放阅读框为1146 bp; AdipoR2 cDNA全长为1590 bp, 开放阅读框为1086 bp。多重序列比对结果显示, 西伯利亚鲟AdipoR1与哺乳类、鸟类、两栖类和鱼类的氨基酸一致性较高, AdipoR2与鱼类氨基酸一致性更高。系统进化树分析显示, 西伯利亚鲟AdipoR1与其他硬骨鱼类聚为一支, AdipoR2与斑点雀鳝聚为一支。实时荧光定量结果显示, AdipoR1和AdipoR2 mRNA在西伯利亚鲟组织中均有表达, AdipoR1在中脑、延脑、前脑、小脑和下丘脑中的表达高于外周组织, AdipoR2在中脑、鳃、瓣肠、性腺和延脑的表达相对较高。此外, 与其他硬骨鱼类似, 西伯利亚鲟AdipoR1和AdipoR2在肌肉中表达较低。当禁食10d时, 西伯利亚鲟幼鱼肌肉中AdipoR1和AdipoR2 mRNA表达量均显著上调, 复投喂后AdipoR1表达显著降低, 而AdipoR2表达无显著变化。由上可知, 西伯利亚鲟AdipoR1和AdipoR2可能参与能量稳态的维持。文章为进一步研究AdipoR1和AdipoR2在肌肉中的作用提供了理论基础。

     

    Abstract: To explore the expression characteristics of Adiponectin receptor and its response to changes of energy state in Siberian sturgeon, the full-length cDNA of AdipoR1 and AdipoR2 were cloned. The total length of AdipoR1 cDNA of Siberian sturgeon was 2013 bp containing an open reading frame of 1146 bp, and the total length of AdipoR2 cDNA was 1590 bp containing an open reading frame of 1086 bp. The results of multiple sequence alignment showed that the amino acids of AdipoR1 were highly consistent with those of mammals, birds, amphibians and fish, while the amino acids of AdipoR2 were highly consistent with those of fish. The phylogenetic tree analysis showed that the Siberian sturgeon AdipoR1 was clustered into the branch with other bony fishes, and Siberian sturgeon AdipoR2 was clustered into the branch with Lepisosteus oculatus. The results of qRT-PCR showed that AdipoR1 and AdipoR2 were widely expresed. AdipoR1 was highly expressed in the midbrain, medulla oblongata, forebrain, cerebellum and hypothalamus of Siberian sturgeon, and AdipoR2 was abundant in the midbrain, gill, valve intestine, gonad and medulla oblongata. Similar to other bony fish, AdipoR1 and AdipoR2 were lowly expressed in muscle of Siberian sturgeon. 10 days fasting significantly induced the expression of AdipoR1 and AdipoR2 mRNA level in muscle of juvenile Siberian sturgeon, and re-feeding significantly decreased the expression of AdipoR1. These results suggest that AdipoR1 and AdipoR2 might be involved in maintaining energy homeostasis in Siberian sturgeon, providing a theoretical basis for further research on the role of AdipoR1 and AdipoR2 in muscle.

     

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