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Ready for the journey: a comparative proteome profiling of porcine cauda epididymal fluid and spermatozoa

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Abstract

In the present study, we describe the proteome of porcine cauda epididymis fluid and spermatozoa by means of Multidimensional Protein Identification Technology (MudPIT). Ten sexually mature healthy boars were surgically castrated and epididymides were dissected to obtain the cauda epididymal content. Polled protein extracts of cauda epididymal fluid (CEF) and spermatozoa (CESperm) were loaded in an Agilent 1100 quaternary HPLC and peptides eluted from the microcapillary column were electro-sprayed directly into a LTQ Orbitrap XL mass spectrometer. Using bioinformatics, identified proteins were classified by their molecular functions, involvement in biological processes and participation in relevant metabolic pathways associated with spermatozoa physiology, fertility potential and protection. A total of 645 proteins were identified in the CEF, with epididymal-specific lipocalin-5, beta-hexosaminidase subunit beta precursor and phosphatidylethanolamine-binding protein 4 being the most abundant proteins found. A total of 2886 proteins were identified in the CESperm proteome with 81 proteins being considered more abundant (spectral counts > 100). CEF and CESperm data were compared and 345 proteins were present in both proteomes. Phosphatidylethanolamine-binding protein 4 precursor was the only protein found most abundant in both CEF and CESperm proteomes. Based on Gene Ontology analysis, we identified CEF and CESperm proteins associated with sperm protection against ROS and immune mediated response, glycosaminoglycan degradation, ubiquitin-proteasome system, metabolic process and maturation, modulation of acrosome reaction and ZP binding and oocyte penetration. These results provide a better comprehension about the molecular process and biological pathways involved in sperm epididymis maturation and establishment of the cauda epididymis sperm reservoir.

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Acknowledgments

The authors wish to thank the Associação de Criadores de Suínos do Rio Grande do Sul (ACSURS) for the provision of reproductive tissue samples. The authors would also like to thank Prof. Dr. Verônica Contini for contributions in the data analysis.

Funding

This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) grant 447251/2014-7, Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS) grant 16/2551-0000223-1 and Fundação Vale do Taquari de Educação e Desenvolvimento Social (FUVATES) grant 08/Reitoria/Univates/2016. Mrs. Laura Espíndola Argenti and Mrs. Ana Paula Binato de Souza were CAPES scholarship recipients.

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Correspondence to Ivan Cunha Bustamante-Filho.

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All procedures with animals described in this article were previously approved by the Local Animal Experimentation Ethics Committee (protocol 001/2015) and according to the Brazilian Animal Welfare legislation.

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Weber, A., Argenti, L.E., de Souza, A.P.B. et al. Ready for the journey: a comparative proteome profiling of porcine cauda epididymal fluid and spermatozoa. Cell Tissue Res 379, 389–405 (2020). https://doi.org/10.1007/s00441-019-03080-0

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Keywords

  • Boar
  • Epididymis
  • Proteomics
  • Spermatozoa
  • Epididymal fluid