大口黑鲈leptin A基因的组织表达及其对急性低氧胁迫的响应

Tissue expression of leptin A gene in Micropterus salmoides and its response to acute hypoxia stress

  • 摘要: 瘦素(leptin)是一种重要的激素,参与调节动物的摄食、繁殖和能量消耗,其可以通过抑制食欲和促进能量代谢的方式维持机体能量稳态。大多数鱼类具有双重瘦素基因。在本研究中,利用PCR技术克隆了大口黑鲈leptin A基因的编码区,其开放阅读框(ORF)为486 bp,编码161个氨基酸蛋白。通过与其他物种进行同源性比对,发现鲈形目的leptin基因的保守性较高,一致性高达91.46%,在大口黑鲈leptin A中几乎不存在基因特异性突变位点,而且大口黑鲈 leptin A氨基酸序列与鳜同源性最高(92.59%),与花鲈(89.51%)、布氏鲳鲹(87.04%)、斜带石斑鱼(83.87%)的同源性次之。组织分布结果显示,leptin A基因在肝脏中的表达量最高,在心脏、头肾、脑、中肾、肠、脾脏、鳃、肌肉和性腺中的表达量均较低。此外,对大口黑鲈进行不同程度急性低氧胁迫重度低氧(1.2±0.2)mg/L和中度低氧(3.5±0.2)mg/L,发现低氧胁迫初期时,严重低氧组和中度低氧组的leptin A的表达量都升高,显著高于对照组 Leptin A 的表达。这表明急性低氧能够诱导大口黑鲈leptin A在肝脏中的表达。综上所述,大口黑鲈leptin A基因与鳜leptin A基因的同源性最高,leptin A主要在大口黑鲈肝脏中显著表达,急性低氧胁迫能显著诱导leptin A的表达。

     

    Abstract: leptin is an important hormone involved in the regulation of feeding, reproduction and energy consumption in animals, and it also can maintain body energy homeostasis by curbing appetite and promoting energy metabolism in vertebrates. Most fishes have the double leptin genes. In this research, the coding region of leptin A was cloned using PCR reaction in Micropterus salmoides, and the results showed that its open reading frame (ORF) was 486 bp, encoding 161 amino acid proteins. Compared with other species, it was found that the conservative property of leptin in Perciformes reached up to 91.46%, there is almost no gene specific mutation site in leptin A of M. salmoides, and the amino acid sequence of leptin A in M. salmoides had the highest homology with the Siniperca chuatsi (92.59%), followed by the Lateolabrax japonicus(89.51%), Trachinotus blochii (87.04%) and Epinephelus coioides (83.87%). Then, leptin A gene was also detected to be highly expressed in liver by fluorescence quantitative PCR (qPCR), subsequently in heart, head kidney, brain, middle kidney, intestine, spleen, gill, muscle and gonad. In addition, acute hypoxic stress was designed to detect the expression of leptin A (1.2±0.2) mg/L and (3.5±0.2) mg/L. The results showed that at the beginning of hypoxia, the expression level of leptin A in the extreme hypoxia group and the moderate hypoxia group was significantly higher than that in the control group, and this indicates that acute hypoxia can induce leptin A expression in liver. In summary, leptin A of M. salmoides has the highest homology with Siniperca chuatsi(92.59%); leptin A was significantly expressed in the liver of M. salmoides; acute hypoxic stress can significantly induce the expression of leptin A.

     

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