Huang Chencui, Yu Kun, Huang Huiyang, Ye Haihui
College of Ocean and Earth Sciences, Xiamen University, Xiamen 361102, People's Republic of China.
J Genet. 2016 Dec;95(4):923-932. doi: 10.1007/s12041-016-0717-z.
Adenosine monophosphate-activated protein kinase (AMPK), an important energy sensor, is crucial for organism survival under adverse conditions. In this study, the roles of this gene under cold stress in a warm-water mud crab, Scylla paramamosain was investigated. The full-length cDNA (SpAMPK) was 1884 bp and its open reading frame of 1566 bp was isolated and characterized. The expressions of SpAMPK detected by quantitative real-time PCR (qRT-PCR) in various tissues revealed that the highest expression was in the hepatopancreas. The profiles of SpAMPK gene in the hepatopancreas, chela muscle and gill were detected when the subadult crabs were exposed to the four temperature conditions of 10, 15, 20 and 25°C. The results showed that the expression patterns of SpAMPK mRNA in the three tissues were significantly higher when crabs were exposed to 15°C than the other three temperature treatments, while at 10°C treatment, the SpAMPK mRNA was lowest among the four temperature treatments. These findings suggested that the high expression of SpAMPK mRNA might initiate ATP-producing pathways to generate energy to cope with cold stress at 15°C treatment, which was slightly below the range of optimum temperatures; while treatment at 10°C, far lower than optima, the low expression of SpAMPK mRNA could reduce the energy expenditure and thus induce the crabs into cold anesthesia. The results of SpAMPK in this study might contribute to the understanding of the molecular mechanism of acclimation to cold hardiness in S. paramamosain.
腺苷单磷酸激活蛋白激酶(AMPK)是一种重要的能量传感器,对生物体在不利条件下的生存至关重要。在本研究中,对该基因在暖水蟹拟穴青蟹冷应激下的作用进行了研究。分离并鉴定了全长1884 bp的cDNA(SpAMPK),其开放阅读框为1566 bp。通过实时定量PCR(qRT-PCR)检测不同组织中SpAMPK的表达,结果显示肝胰腺中的表达最高。当亚成体蟹暴露于10、15、20和25°C这四种温度条件下时,检测了肝胰腺、螯肌和鳃中SpAMPK基因的表达情况。结果表明,当蟹暴露于15°C时,这三种组织中SpAMPK mRNA的表达模式显著高于其他三种温度处理,而在10°C处理时,SpAMPK mRNA在四种温度处理中最低。这些发现表明,SpAMPK mRNA的高表达可能启动产生ATP的途径以产生能量,从而应对15°C处理下的冷应激,该温度略低于最适温度范围;而在10°C处理时,远低于最适温度,SpAMPK mRNA的低表达可减少能量消耗,从而诱导蟹进入冷麻醉状态。本研究中SpAMPK的结果可能有助于理解拟穴青蟹适应耐寒性的分子机制。