Cloning of Mn-SOD gene and its mRNA expression difference and antioxidant enzyme activities under hypoxia stress of cobia Rachycentron canadum

Background Environmental hypoxia affects the survival and development of organisms. It is also an important environmental factor that leads to oxidative damage. Hypoxia is a condition in which tissues are deprived of oxygen; reoxygenation is the phenomenon in which hypoxic tissues are exposed to oxygen. Hypoxia-reoxygenation is vital in pathogenesis, where the production of reactive oxygen species and antioxidant disparity significantly contribute to disease progression, and it is one of the most common physiological stressors in the aquaculture industry. Methods and results In this study, the full length of complementary DNA (cDNA) of the manganese superoxide dismutase (Mn-SOD) gene of healthy cobia Rachycentron canadum was analysed using rapid amplification of cDNA ends. The real-time quantitative Polymerase Chain Reaction was used to measure the expression levels of Mn-SOD mRNAs in various tissues (heart, muscle, brain, liver, kidney, gill, intestine, and spleen). The 2(-Delta Delta CT) method was used to performed the expression analysis. The experimental data were analysed using SPSS ver. 19.0 (). P < 0.05 and P < 0.01 were set as significant differences. The values were articulated as mean +/- standard deviation. The Mn-SOD gene cDNA sequence was 1209 bp long, including a 684 bp open reading frame, 42 bp 5'UTR and 483 bp 3'UTR, encoding 227 amino acids. Under hypoxia-reoxygen stress, the expression of Mn-SOD in brain tissue was significantly lower than in the control group after 8 h of reoxygenation and higher than the control group after 24 h. Hypoxia and subsequent reoxygenation triggered a disturbance in antioxidant homeostasis, displayed in the modification of GPx expression/activity in the liver: GPx was improved. Conclusions These results provide valuable information on the role of Mn-SOD regulation in oxidative stress caused by hypoxia.

Automated summary

This study analyzed the cDNA sequence of the Mn-SOD gene in cobia and found that the expression of Mn-SOD in brain tissue was significantly affected under hypoxia-reoxygenation stress. Additionally, changes in GPx expression/activity in the liver indicated disturbance in antioxidant homeostasis. These results provide valuable information on the role of Mn-SOD regulation in oxidative stress caused by hypoxia.