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Mercury-Induced Inhibition of Tyrosine Phosphorylation of Sperm Proteins and Altered Functional Dynamics of Buck Spermatozoa: an In Vitro Study

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Present study was undertaken on buck spermatozoa to investigate the effect of mercuric chloride on functional dynamics of buck spermatozoa. Four different concentrations (0.031, 0.125, 0.25 and 1.25 μg/mL) of mercuric chloride, which were 1/40th, 1/10th, 1/5th and equivalent to the LC50 value of HgCl2, were selected for studying their effect following in vitro exposure for 15 min and 3 h. Exposure of spermatozoa to 0.031 μg/mL mercuric chloride for 3 h resulted in significant (p < 0.05) decrease in sperm motility, sperm having intact membrane, intact acrosome and high mitochondrial trans-membrane potential. However, following exposure to higher concentrations (0.25, 1.25 μg/mL), similar results were observed even after 15 min of exposure. HgCl2 significantly (p < 0.05) increased the levels of malondialdehyde and reactive oxygen species and significantly (p < 0.05) decreased total antioxidant capacity and superoxide dismutase activity in spermatozoa within 15 min of exposure. Mercuric chloride-treated spermatozoa did not show capacitation, rather exhibited spontaneous acrosome reaction along with significant increase in intracellular Ca2+ and cAMP levels. Immuno-blotting of semen samples of control and 0.031 μg/mL mercury-treated groups showed low intensity bands of p55, p70, p80, p105 and p190 kDa tyrosine phosphorylation proteins while higher concentration-treated groups showed no such bands. Our findings evidently suggest that mercuric chloride even at 0.031 μg/mL adversely affected sperm functions, inhibited tyrosine phosphorylation proteins and capacitation due to oxidative stress. Spontaneous acrosome reaction (AR) in mercury-treated spermatozoa may possibly be due to increase in intracellular Ca2+ and cAMP levels, and capacitation failure may be due to inhibition of tyrosine phosphorylation of proteins.

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Acknowledgements

We are also thankful to Professor and Head, Department of Pharmacology and Toxicology, for providing the necessary facilities in ICAR Niche Area of Excellence Laboratories. Laboratory facilities provided by the former and present Heads, Department of Veterinary Physiology, and technical assistance provided by Dr. Brijesh Yadav and Dr. Mukul Anand are also thankfully acknowledged.

Funding

Financial assistance was provided by Dean, College of Biotechnology, to carry out this work.

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Authors and Affiliations

  1. College of Biotechnology, U.P. Pandit Deendayal Upadhayaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, Uttar Pradesh, 281001, India

    Bhawna Kushawaha

  2. Department of Pharmacology and Toxicology, U.P. Pandit Deendayal Upadhayaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, Uttar Pradesh, 281001, India

    Rajkumar Singh Yadav & Satish Kumar Garg

  3. Department of Veterinary Physiology, U.P. Pandit Deendayal Upadhayaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, Uttar Pradesh, 281001, India

    Dilip Kumar Swain

  4. BD Bioscience, Gurgaon, India

    Pradeep K Rai

  5. College of Veterinary Science and Animal Husbandry, U.P. Pandit Deendayal Upadhayaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, Uttar Pradesh, 281001, India

    Bhawna Kushawaha, Rajkumar Singh Yadav, Dilip Kumar Swain & Satish Kumar Garg

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The guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals Govt. of India were followed as per the approval of Institutional Animal Ethics Committee (Approval No. 110/IAEC/16, dated: 16-09-2016).

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Kushawaha, B., Yadav, R.S., Swain, D.K. et al. Mercury-Induced Inhibition of Tyrosine Phosphorylation of Sperm Proteins and Altered Functional Dynamics of Buck Spermatozoa: an In Vitro Study. Biol Trace Elem Res 198, 478–492 (2020). https://doi.org/10.1007/s12011-020-02077-z

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