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

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Abstract

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–ΔΔCT method was used to performed the expression analysis. The experimental data were analysed using SPSS ver. 19.0 (https://spss.software.informer.com/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.

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Data availability

Upon reasonable demand, the data supporting the results of this study are obtainable from the corresponding author.

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Acknowledgements

Our acknowledgment goes to the funders of this study.

Funding

This work was supported by grants from China Agriculture Research System (CARS-47); Southern Marine Science and Engineering Guangdong Laboratory [(Zhanjiang) (ZJW-2019-06)]; Guangdong Ocean University Undergraduate Innovation and Entrepreneurship Training Programs (CXXL2018131).

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    Authors and Affiliations

    1. Fishery College, Guangdong Ocean University, Zhanjiang, 524025, China

      Jian-Dong Zhang, Hong-Juan Li, Eric Amenyogbe, Wei-Zheng Wang, Jian-Sheng Huang & Gang Chen

    2. Southern Marine Science and Engineering Guangdong Laboratory (Zhanjiang), Zhanjiang, 524025, China

      Jian-Dong Zhang, Jian-Sheng Huang & Gang Chen

    3. Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, 524088, China

      Jian-Dong Zhang, Jian-Sheng Huang & Gang Chen

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    1. Jian-Dong Zhang
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    2. Hong-Juan Li
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    Contributions

    Jian-dong Zhang was responsible for experiment design, data processing, and the writing; HJL was responsible for experimental sampling and gene cloning. WZW was responsible for sampling and gene expression. EA is responsible for writing and revising the manuscript. GC is responsible for experiment design and the culture of experimental fish. JSH was responsible for funding.

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    Correspondence to Jian-Sheng Huang or Gang Chen.

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    All procedures were executed in strict accordance with the regulations for the administration of laboratory animals in Guangdong province, China, and the Guangdong Ocean University Research Council’s guide for the care and use of laboratory Animals. All experimental processes were approved by the Guangdong Ocean University Research Council. With approval number (GDOU-LAE-2020-013).

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    Zhang, JD., Li, HJ., Amenyogbe, E. et al. Cloning of Mn-SOD gene and its mRNA expression difference and antioxidant enzyme activities under hypoxia stress of cobia Rachycentron canadum. Mol Biol Rep 48, 6897–6909 (2021). https://doi.org/10.1007/s11033-021-06692-4

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