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Role of the bovine PRAMEY protein in sperm function during in vitro fertilization (IVF)

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Abstract

Preferentially expressed antigen in melanoma (PRAME) is a cancer/testis antigen (CTA) that is predominantly expressed in normal male gonad tissues and a variety of tumors. PRAME proteins are present in the acrosome and sperm tail, but their role in sperm function is unknown. The objective of this study was to examine the function of the bovine Y-linked PRAME (PRAMEY) during spermatozoal capacitation, the acrosome reaction (AR), and fertilization. Freshly ejaculated spermatozoa were induced to capacitate and undergo AR in vitro. Western blotting results revealed a decrease in the PRAMEY protein in capacitated spermatozoa, and the release of the PRAMEY protein from the acrosome during the AR, suggesting its involvement in sperm capacitation and AR. IVF was performed using in vitro matured bovine oocytes and cauda epididymal spermatozoa either treated with PRAMEY antibody, rabbit IgG, or DPBS. Sperm-egg binding and early embryos were examined at 6 and 45 h post IVF, respectively. The number of spermatozoa that bound per oocyte was nearly two-fold greater in the PRAMEY antibody treatment group (34.4) when compared to both the rabbit IgG (17.6) and DPBS (18.1) controls (P < 0.01). Polyspermy rate in the antibody-treated group (18.9%) was three-fold greater than the rabbit IgG control (6.0%) (P < 0.01). The results indicate that PRAMEY may play a role in anti-polyspermy defense. This study thus provides the initial evidence for the involvement of the PRAME protein family in sperm function and fertilization.

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Acknowledgements

The authors would like to thank Mr. Duane Eichenlaub and Mr. Douglas Nicholas at Nicholas Meat, LLC (Loganton, PA) for providing bovine testes and ovaries. The authors are also grateful to Dr. Bo Harstine at Select Sires, Inc. for providing fresh bovine semen samples.

Funding

This work is supported by the National Institute of Food and Agriculture (NIFA), United States Department of Agriculture (USDA), grant no. 2018–67015-27576 (WSL) and grant no. 2021–67015-33404 (PS).

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Author notes

  1. Chen Lu

    Present address: Fudan University, Shanghai, People’s Republic of China

  2. Yaqi Zhao

    Present address: St. Jude Children’s Research Hospital, Memphis, TN, USA

  3. Olga Maria Ocon-Grove

    Present address: Actuated Medical, Inc., PA, Bellefonte, USA

Authors and Affiliations

  1. Department of Animal Science, Center for Reproductive Biology and Health (CRBH), College of Agricultural Sciences, The Pennsylvania State University, 311 AVBS Building, University Park, PA, 16802, USA

    Chandlar Kern, Weiwei Wu, Chen Lu, Jianbin Zhang, Yaqi Zhao, Olga Maria Ocon-Grove, Francisco Diaz & Wan-Sheng Liu

  2. Animal Science Institute, Xinjiang Academy of Agriculture Science, Xinjiang, China

    Weiwei Wu

  3. Department of Animal Science, Tianjin Agriculture University, Tianjin, China

    Jianbin Zhang

  4. Division of Animal Sciences, and Department of Obstetrics, Gynecology & Women’s Health, University of Missouri, Columbia, MO, USA

    Peter Sutovsky

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  1. Chandlar Kern
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Contributions

WL conceived and designed the research project. CK, WW, CL, JZ, YZ, OMOG, PS, and FD participated in the IVF experimental design. CK, WW, CL, JZ, and YZ carried out IVF experiments. CK, WW, CL, JZ, YZ, OMOG, PS, FD, and WL contributed to the interpretation of the IVF results. CK carried out sperm capacitation and acrosome reaction experiments and data analysis. PS performed PRAMEY/PNA IF staining on spermatids/spermatozoa. CK took the lead in writing the manuscript. WL and PS provided critical feedback and helped shape the research, analysis, and manuscript. All authors approved the manuscript.

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Correspondence to Wan-Sheng Liu.

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Kern, C., Wu, W., Lu, C. et al. Role of the bovine PRAMEY protein in sperm function during in vitro fertilization (IVF). Cell Tissue Res 391, 577–594 (2023). https://doi.org/10.1007/s00441-022-03717-7

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