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Organic Zinc and Copper Supplementation–Associated Changes in Gene Expression and Protein Profiles in Buck Spermatozoa

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Abstract

Mineral supplementation has greater impact on male reproduction; however, the mechanism of action has not been studied in detail. The present study was aimed to deal with the lacuna in mechanism of action of mineral supplementation on improvement in sperm characteristics. A group of 40 bucks (aged 5 months) were assigned to 10 groups (4 in each group) based on their body weight and fed with concentrate mixture: basal roughage (minimal diet) in equal proportion to all the bucks. Among the 10 groups, one was considered as control, without any additional mineral supplementation, and the remaining 9 were treatment groups (3 groups each in Zn, Cu, and Zn + Cu). In treatment groups, organic Zn was fed in three different doses as 20, 40, and 60 mg/kg DM; organic Cu was fed in three different doses as 12.5, 25, and 37.5 mg/kg DM; and organic Zn + Cu was combinedly supplied as 20 + 12.5, 40 + 25, and 60 + 37.5 based on their mg/kg DM for 8 months period. The neat semen samples were processed for spermatozoal gene (stress- NOS3, HSP70, HIF1A; fertility- MTF1, MTA1, TIMP2, TNFa, and EGFR) expression studies through qRT-PCR and protein profile changes through single- and two-dimensional gel electrophoresis. Significantly, the stress-responsive genes were downregulated, and fertility-related genes were upregulated in treatment groups. A significant correlation had been noticed among the genes studied: HIF1A with MTA1 (P < 0.05) and MTF1 with EGFR, TIMP2, TNFa, and NOS3 (P < 0.01) respectively. The organic Zn and Cu feeding modulated the expression of stress- and fertility-related genes and protein abundance, thereby improved the sperm characteristics.

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Data Availability

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Acknowledgements

We acknowledge the Department of Biotechnology, Jain University, Bengaluru, India, for helping to register the doctoral program; CSIR HRDG (Council of Scientific & Industrial Research), Government of India, for providing the CSIR fellowship to do Ph.D; The Director, ICAR-NIANP, Bengaluru, for providing research workspace and facilities at the institute; and The Director, DBT, Government of India, for supporting the research work through chemicals and kits.

Funding

This research work was funded by DBT, Ministry of Science & Technology, Government of India (BT/PR10901/AAQ/1/581/2014) and CSIR HRDG, Government of India.

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VM conducted the research and prepared the manuscript; PG helped in statistical analysis of the data. AA designed the experiment and helped for preparing the article; SS helped in designing the work; JG assisted in preparing and proof reading the manuscript.

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Correspondence to Arangasamy Arunachalam.

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This study was carried out in the Reproductive Physiology Laboratory at ICAR-NIANP, Bengaluru, with necessary approval from institutional animal ethics committee (IAEC).

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Mayasula, V.K., Arunachalam, A., Sellappan, S. et al. Organic Zinc and Copper Supplementation–Associated Changes in Gene Expression and Protein Profiles in Buck Spermatozoa. Biol Trace Elem Res 200, 1626–1639 (2022). https://doi.org/10.1007/s12011-021-02796-x

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