Skip to main content

Advertisement

SpringerLink
Log in

SERPINA5 Protein in Cumulus-Oocyte Complexes Increases the Fertilisation Ability of Mouse Sperm

  • Reproductive Biology.: Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

Obtaining high-quality sperm is key to improving the success rate of assisted reproductive technology (ART). Although cytokines secreted by cumulus-oocyte complexes (COCs) bind to sperm surface receptors to improve sperm quality, the effects of adding mouse COCs to human tubal fluid (HTF) medium on sperm capacitation have not yet been explored. Eight-week-old ICR mouse COCs were added to HTF medium and crushed to obtain the post-modified HTF medium. Compared with using HTF medium, the fertilisation rate and number of sperm combined with the zona pellucida significantly increased after in vitro capacitation using the post-modified HTF medium (P < 0.01). Proteomic and Western blotting analyses showed that the level of SERPINA5 in sperm increased significantly following in vitro capacitation with the post-modified HTF medium (P < 0.05). Immunohistochemical staining analysis demonstrated that SERPINA5 protein was expressed in mouse cumulus cells. A SERPINA5 antibody was added in the post-modified HTF medium to block the effects of SERPINA5 after in vitro capacitation, which significantly decreased the fertilisation rate and the number of sperm combined with the zona pellucida (P < 0.05). Recombinant mouse SERPINA5 protein (1 ~ 2 μg/ml) was added to HTF medium and the fertilisation rate and the number of sperm combined with the zona pellucida significantly increased (P < 0.01). Moreover, recombinant human SERPINA5 protein (5 μg/ml) was added before human semen freezing. Compared with adding no SERPINA5 protein, the percentage of normal sperm morphology and the intact acrosome significantly increased (P < 0.05). Our study provides a reference method for optimising sperm quality in the process of in vitro capacitation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Data Availability

All data generated or analysed during this study are included in this published article and its supplementary information files.

Code Availability

Not applicable.

References

  1. Steptoe PC, Edwards RG. Birth after the reimplantation of a human embryo. Lancet. 1978;2(8085):366.

    Article  CAS  Google Scholar 

  2. Moore SG, Hasler JF. A 100-year review: reproductive technologies in dairy science. J Dairy Sci. 2017;100(12):10314–31.

    Article  CAS  Google Scholar 

  3. De Geyter C. Assisted reproductive technology: impact on society and need for surveillance. Best Pract Res Clin Endocrinol Metab. 2019;33(1):3–8.

    Article  Google Scholar 

  4. Nasr-Esfahani MH, Deemeh MR, Tavalaee M. New era in sperm selection for ICSI. Int J Androl. 2012;35(4):475–84.

    Article  CAS  Google Scholar 

  5. Yan Y, Liu H, Zhang B, Liu R. A PMMA-based microfluidic device for human sperm evaluation and screening on swimming capability and swimming persistence. Micromachines (Basel). 2020;11(9):793.

    Article  Google Scholar 

  6. Vaughan DA, Sakkas D. Sperm selection methods in the 21st century. Biol Reprod. 2019;101(6):1076–82.

    Article  Google Scholar 

  7. Lepine S, McDowell S, Searle LM, Kroon B, Glujovsky D, Yazdani A. Advanced sperm selection techniques for assisted reproduction. Cochrane Database Syst Rev. 2019;7(7):CD010461.

  8. Puga Molina LC, Luque GM, Balestrini PA, Marin-Briggiler CI, Romarowski A, Buffone MG. Molecular basis of human sperm capacitation. Front Cell Dev Biol. 2018;6:72.

    Article  Google Scholar 

  9. Moreno-Irusta A, Kembro JM, Dominguez EM, Matamoros-Volante A, Gallea MN, Molina R, et al. Sperm physiology varies according to ultradian and infradian rhythms. Sci Rep. 2019;9(1):5988.

    Article  Google Scholar 

  10. Cormier N, Bailey JL. A differential mechanism is involved during heparin- and cryopreservation-induced capacitation of bovine spermatozoa. Biol Reprod. 2003;69(1):177–85.

    Article  CAS  Google Scholar 

  11. Kulanand J, Shivaji S. Capacitation-associated changes in protein tyrosine phosphorylation, hyperactivation and acrosome reaction in hamster spermatozoa. Andrologia. 2001;33(2):95–104.

    Article  CAS  Google Scholar 

  12. Umezu K, Hara K, Hiradate Y, Numabe T, Tanemura K. Stromal cell-derived factor 1 regulates in vitro sperm migration towards the cumulus-oocyte complex in cattle. PLoS One. 2020;15(4):e0232536.

    Article  CAS  Google Scholar 

  13. Sarapik A, Velthut A, Haller-Kikkatalo K, Faure GC, Béné M-C, de Carvalho Bittencourt M, et al. Follicular proinflammatory cytokines and chemokines as markers of IVF success. Clin Dev Immunol. 2012;2012:606459.

  14. Sugimoto Y, Inazumi T, Tsuchiya S. Roles of prostaglandin receptors in female reproduction. J Biochem. 2015;157(2):73–80.

    Article  CAS  Google Scholar 

  15. Diao R, Wang T, Fok KL, Li X, Ruan Y, Yu MK, et al. CCR6 is required for ligand-induced CatSper activation in human sperm. Oncotarget. 2017;8(53):91445–58.

    Article  Google Scholar 

  16. Sabet S, Najafi MH, Tavalaee M, Sadeghi N, Nasr-Esfahani MH. Single-blind clinical trial: sperm selection based on capacity to pass through cumulus oophorus column improves ICSI outcomes. Andrology. 2021;9(5):1560–70.

    Article  CAS  Google Scholar 

  17. Wahlmuller FC, Yang H, Furtmuller M, Geiger M. Regulation of the extracellular SERPINA5 (protein C inhibitor) penetration through cellular membranes. Adv Exp Med Biol. 2017;966:93–101.

    Article  Google Scholar 

  18. Panner Selvam MK, Agarwal A, Pushparaj PN, Baskaran S, Bendou H. Sperm proteome analysis and identification of fertility-associated biomarkers in unexplained male infertility. Genes (Basel). 2019;10(7):522.

    Article  Google Scholar 

  19. Elisen MG, Van Kooij RJ, Nolte MA, Marquart JA, Lock TM, Bouma BN, et al. Protein C inhibitor may modulate human sperm-oocyte interactions. Biol Reprod. 1998;58(3):670–7.

    Article  CAS  Google Scholar 

  20. Hayashi KG, Ushizawa K, Hosoe M, Takahashi T. Differential gene expression of serine protease inhibitors in bovine ovarian follicle: possible involvement in follicular growth and atresia. Reprod Biol Endocrinol. 2011;9:72.

    Article  CAS  Google Scholar 

  21. Uhrin P, Dewerchin M, Hilpert M, Chrenek P, Schöfer C, Zechmeister-Machhart M, et al. Disruption of the protein C inhibitor gene results in impaired spermatogenesis and male infertility. J Clin Invest. 2000;106(12):1531–9.

    Article  CAS  Google Scholar 

  22. Ma J, Chen Q, Wang S, Ma R, Jing J, Yang Y, et al. Mitochondria-related miR-574 reduces sperm ATP by targeting ND5 in aging males. Aging (Albany NY). 2020;12(9):8321–38.

    Article  CAS  Google Scholar 

  23. Nakao S, Takeo T, Watanabe H, Kondoh G, Nakagata N. Successful selection of mouse sperm with high viability and fertility using microfluidics chip cell sorter. Sci Rep. 2020;10(1):8862.

    Article  CAS  Google Scholar 

  24. Soleimanzadeh A, Talavi N, Yourdshahi VS, Bucak MN. Caffeic acid improves microscopic sperm parameters and antioxidant status of buffalo (Bubalus bubalis) bull semen following freeze-thawing process. Cryobiology. 2020;95:29–35.

    Article  CAS  Google Scholar 

  25. Quinn MM, Jalalian L, Ribeiro S, Ona K, Demirci U, Cedars MI, et al. Microfluidic sorting selects sperm for clinical use with reduced DNA damage compared to density gradient centrifugation with swim-up in split semen samples. Hum Reprod. 2018;33(8):1388–93.

    Article  CAS  Google Scholar 

  26. Macias-Garcia B, Gonzalez-Fernandez L, Loux SC, Rocha AM, Guimarães T, Peña FJ, et al. Effect of calcium, bicarbonate, and albumin on capacitation-related events in equine sperm. Reprod. 2015;149(1):87–99.

    Article  CAS  Google Scholar 

  27. Maciel VL Jr, Caldas-Bussiere MC, Silveira V, Reis RS, Rios AFL, Paes de Carvalho CS. L-arginine alters the proteome of frozen-thawed bovine sperm during in vitro capacitation. Theriogenology. 2018;119:1–9.

    Article  CAS  Google Scholar 

  28. Silva EC, Cajueiro JF, Silva SV, Soares PC, Guerra MMP. Effect of antioxidants resveratrol and quercetin on in vitro evaluation of frozen ram sperm. Theriogenology. 2012;77(8):1722–6.

    Article  CAS  Google Scholar 

  29. Salustri A, Camaioni A, Di Giacomo M, Fulop C, Hascall VC. Hyaluronan and proteoglycans in ovarian follicles. Hum Reprod Update. 1999;5(4):293–301.

    Article  CAS  Google Scholar 

  30. España F, Sánchez-Cuenca J, Fernández PJ, Gilabert J, Romeu A, Estellés A, Royo M, Muller CH. Inhibition of human sperm-zona-free hamster oocyte binding and penetration by protein C inhibitor. Andrologia. 1999;31(4):217–23.

    Article  Google Scholar 

  31. Wagenaar GT, Uhrin P, Weipoltshammer K, Almeder M, Hiemstra PS, Geiger M, et al. Expression patterns of protein C inhibitor in mouse development. J Mol Histol. 2010;41(1):27–37.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the other members of Dr. Yao’s laboratory for their discussion and help.

Funding

This work was funded by the National Key Research and Development Program of China [grant no. 2018YFC1004700], the Key Research and Development Program of Jiangsu Province [grant no. BE2018714], the National Natural Science Foundation of China [grant nos. 81971373, 81701431, 81901547, 82001618], the Natural Science Foundation of Jiangsu Province [BK20190252, BK20170620], the China Postdoctoral Science Foundation [2017M613434], the Foundation for Key Medical Talents in Jiangsu Province [grant no. ZDRCA2016096], and the 333 High-level Personnel Training Project of Jiangsu Province [grant no. BRA2019109].

Author information

Authors and Affiliations

  1. Center of Reproductive Medicine, Nanjing Jinling Hospital, Clinical School of Medical College, Nanjing University, Nanjing, 210002, Jiangsu, China

    Siyuan Cao, Zhang Qian, Ronghua Wu, Shanshan Sun, Jun Jing, Guangyun Zhang, Weiqing Chen, Yifeng Ge, Jinzhao Ma, Shuxian Wang, Lei Ouyang, Tongmin Xue, Dandan Wang, Shanmeizi Zhao, Kuan Liang, Xie Ge, Ying Lin, Li Chen & Bing Yao

  2. School of Life Science, Nanjing Normal University, Nanjing, 210002, Jiangsu, China

    Siyuan Cao, Shanshan Sun & Guangyun Zhang

Authors
  1. Siyuan Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhang Qian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ronghua Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shanshan Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jun Jing
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Guangyun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Weiqing Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yifeng Ge
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jinzhao Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Shuxian Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Lei Ouyang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Tongmin Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Dandan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Shanmeizi Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Kuan Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Xie Ge
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Ying Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. Li Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Bing Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Bing Yao, Siyuan Cao, Jinzhao Ma, Yifeng Ge, Zhang Qian and Ronghua Wu conceived and designed the study. All authors participated in the study and analysed and interpreted the data. Bing Yao, Jinzhao Ma and Jun Jing revised the article for important intellectual content. Siyuan Cao and Zhang Qian wrote the article. All the authors approved the final version of the manuscript.

Corresponding author

Correspondence to Bing Yao.

Ethics declarations

Ethics Approval and Consent to Participate

This study was approved by the ethics committee of Nanjing Jinling Hospital. All animal experiments were approved by the Animal Protection and Use Committee of Jinling Hospital and carried out in accordance with institutional guidelines (2020JLHGKJDWLS-173).

Consent for Publication

Not applicable.

Conflict of Interest

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Siyuan Cao, Zhang Qian and Ronghua Wu are regarded as joint first authors.

Supplementary Information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Cao, S., Qian, Z., Wu, R. et al. SERPINA5 Protein in Cumulus-Oocyte Complexes Increases the Fertilisation Ability of Mouse Sperm. Reprod. Sci. 29, 2350–2362 (2022). https://doi.org/10.1007/s43032-022-00867-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43032-022-00867-5

Keywords

Search

Navigation