Abstract
Copper (Cu) is an essential and important trace element for some significant life processes for most organisms. However, an excessive amount of Cu can be highly toxic. The present study was conducted to elucidate the oxidative stress–induced alteration in transcriptional level of autophagy-related genes in the liver and kidney tissue of fish Channa punctatus after treatment with three different sublethal concentrations of CuSO4 for 28 days. All the studied enzymatic and non-enzymatic oxidative stress markers viz. superoxide dismutase-SOD, catalase-CAT, glutathione peroxidase-GPx, glutathione reductase-GR, and glutathione-GSH showed an increase in their activity levels in the treated groups in a dose-dependent manner. Particularly SOD and CAT have shown a significant hike in activity levels. ROS levels in blood cells increased significantly (p < 0.05) in all the treated groups, i.e., Group II-1/20th of 96 h-LC50 (0.2 mg/L), Group III-1/10th of 96 h-LC50 (0.4 mg/L), and Group IV-1/5 h of 96 h-LC50 (0.8 mg/L) of Cu2+ in a dose-dependent manner as compared to control (Group I). The upregulation in mRNA levels of autophagy-related genes Microtubule-associated protein 1 light chain 3 (LC3), Gamma-aminobutyric acid receptor-associated protein precursor (Gabarap), and Golgi-associated ATPase enhancer of 16 kDa (GATE16), autophagy-related 5 (ATG5) was observed while mammalian target of rapamycin (mTOR) showed downregulation in the liver and kidney tissue of fish. The decrease in mTOR and increase in ATG5 gene expression projects autophagic vesicle formation due to oxidative stress. There was significant induction in micronuclei (MN) frequency in all the treated groups. The highest frequency of MN induced by Cu2+ was recorded in Group IV after 28 days of the exposure period. Thus, it can be concluded that the available information about Cu2+-induced oxidative stress–mediated autophagy in the liver and kidney of fish C. punctatus remains largely unclear to date, so to fill the aforesaid gap, the present study was undertaken, which gives an insight for the mechanisms of autophagy induced by Cu2+ in fish.
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Acknowledgements
We record our sincere thanks to the Department of Higher Education, Uttar Pradesh Government, U.P. India, for providing the Center of Excellence project 2019-20 vide grant no. 66/2019/1864/Satter-4-2019-4(24). We are also grateful to the Head, Department of Zoology, University of Lucknow, Lucknow (226007) for providing laboratory facilities.
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This work was supported by the Department of Higher Education, Uttar Pradesh Government, U.P. India, vide grant no. 66/2019/1864/Satter-4–2019-4(24) and Dr. Manoj Kumar has received research support from the Department of Higher Education, Uttar Pradesh Government.
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All authors contributed to the study’s conception and design. Experimental setup, data collection, and analysis were performed by Dr. Manoj Kumar, Ms. Shefalee Singh, Ms. Shikha Dwivedi, Dr. Abha Trivedi, Dr. Indrani Dubey, and Dr. Sunil P. Trivedi. The first draft of the manuscript was written by Dr. Manoj Kumar and Ms. Shefalee Singh and all authors commented on and revised the first draft of the manuscript. All authors read and approved the final manuscript.
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Under the provisions of the committee for the purpose of control and supervision of experiments on animals (CPCSEA), the Government of India, an Institutional Animal Ethics Committee (IAEC) vide registration no. 1861/GO/Re/S/16/CPCSEA already exists in the University of Lucknow, Lucknow. We have followed the protocols mentioned therein CPCSEA for maintenance and experiment.
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Highlights
• Cu2+ induces oxidative damage due to increased SOD, CAT, GSH, GR, and Gpx in liver and kidney tissues.
• Cu2+ downregulates expression of mTOR gene and promotes autophagy in liver and kidney tissues.
• A disturbed expression of various autophagy marker genes (LC3, ATG5, GABARAP, GATE16) after chronic exposure of Cu2+.
• Exposure of Channa punctatus to Cu2+ shows micronuclei formation in erythrocytes.
• A significant relation was observed between Cu2+-induced oxidative stress and autophagy.
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Kumar, M., Singh, S., Dwivedi, S. et al. Copper-induced Genotoxicity, Oxidative Stress, and Alteration in Transcriptional Level of Autophagy-associated Genes in Snakehead Fish Channa punctatus. Biol Trace Elem Res 201, 2022–2035 (2023). https://doi.org/10.1007/s12011-022-03301-8
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DOI: https://doi.org/10.1007/s12011-022-03301-8