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The surface proteome of bovine unsexed and sexed spermatozoa

(2025) ANIMALS. 15(4).
Author
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Abstract
Current sperm sexing methods are costly and largely restricted to cattle, while immunological techniques targeting sex-specific membrane proteins may offer more economical alternatives. To advance these methods, understanding the proteomic differences between the cell membranes of X- and Y-chromosome-bearing spermatozoa is essential. This study aimed to characterize the cell surface proteome of bovine sperm and identify potential targets for sperm sexing through LC-MS/MS analysis. Cell surface protein lysates were extracted from unsexed, X-sperm (BX), and Y-sperm (BY) samples via biotinylation. Promising targets were identified through functional annotation (UniProt, eggNOG-mapper v.2.1.7) and topology prediction (DeepTMHMM v.1.0.13). Additionally, statistical overrepresentation (PANTHER 18.0) and orthology analyses were performed. Excluding contaminants, 130 proteins were detected, of which 64 proteins were detected in the BX samples and not in the BY samples. Of these, five transmembrane proteins stood out as potential X-sperm targets (ADAM2, ATP11C, DG1, MCT1, and PMCA4). They were identified as potential cell surface targets, based on GO terms and topology predictions, detected in at least two replicates of the BX samples, and shown to share orthology with other livestock species. These findings enhance our understanding of bovine sperm proteomics; however, further validation is required to confirm the utility of these five proteins in sperm sexing technologies.
Keywords
proteomics, sperm surface proteome, sex-specific membrane proteins, sperm sexing, P-TYPE ATPASES, SPERM MOTILITY, PROTEINS, IDENTIFICATION, FERTILIZATION, ATP11C, EXPRESSION, BASIGIN, MOUSE, DAIRY

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MLA
Pinto-Pinho, Patrícia, et al. “The Surface Proteome of Bovine Unsexed and Sexed Spermatozoa.” ANIMALS, vol. 15, no. 4, 2025, doi:10.3390/ani15040484.
APA
Pinto-Pinho, P., Quelhas, J., Impens, F., Dufour, S., Van Haver, D., Lopes, G., … Colaço, B. (2025). The surface proteome of bovine unsexed and sexed spermatozoa. ANIMALS, 15(4). https://doi.org/10.3390/ani15040484
Chicago author-date
Pinto-Pinho, Patrícia, Joana Quelhas, Francis Impens, Sara Dufour, Delphi Van Haver, Graça Lopes, António Rocha, Rosário Pinto-Leite, Margarida Fardilha, and Bruno Colaço. 2025. “The Surface Proteome of Bovine Unsexed and Sexed Spermatozoa.” ANIMALS 15 (4). https://doi.org/10.3390/ani15040484.
Chicago author-date (all authors)
Pinto-Pinho, Patrícia, Joana Quelhas, Francis Impens, Sara Dufour, Delphi Van Haver, Graça Lopes, António Rocha, Rosário Pinto-Leite, Margarida Fardilha, and Bruno Colaço. 2025. “The Surface Proteome of Bovine Unsexed and Sexed Spermatozoa.” ANIMALS 15 (4). doi:10.3390/ani15040484.
Vancouver
1.
Pinto-Pinho P, Quelhas J, Impens F, Dufour S, Van Haver D, Lopes G, et al. The surface proteome of bovine unsexed and sexed spermatozoa. ANIMALS. 2025;15(4).
IEEE
[1]
P. Pinto-Pinho et al., “The surface proteome of bovine unsexed and sexed spermatozoa,” ANIMALS, vol. 15, no. 4, 2025.
@article{01JR7SR4KTZDCECCFZTD418VMJ,
  abstract     = {{Current sperm sexing methods are costly and largely restricted to cattle, while immunological techniques targeting sex-specific membrane proteins may offer more economical alternatives. To advance these methods, understanding the proteomic differences between the cell membranes of X- and Y-chromosome-bearing spermatozoa is essential. This study aimed to characterize the cell surface proteome of bovine sperm and identify potential targets for sperm sexing through LC-MS/MS analysis. Cell surface protein lysates were extracted from unsexed, X-sperm (BX), and Y-sperm (BY) samples via biotinylation. Promising targets were identified through functional annotation (UniProt, eggNOG-mapper v.2.1.7) and topology prediction (DeepTMHMM v.1.0.13). Additionally, statistical overrepresentation (PANTHER 18.0) and orthology analyses were performed. Excluding contaminants, 130 proteins were detected, of which 64 proteins were detected in the BX samples and not in the BY samples. Of these, five transmembrane proteins stood out as potential X-sperm targets (ADAM2, ATP11C, DG1, MCT1, and PMCA4). They were identified as potential cell surface targets, based on GO terms and topology predictions, detected in at least two replicates of the BX samples, and shown to share orthology with other livestock species. These findings enhance our understanding of bovine sperm proteomics; however, further validation is required to confirm the utility of these five proteins in sperm sexing technologies.}},
  articleno    = {{484}},
  author       = {{Pinto-Pinho, Patrícia and Quelhas, Joana and Impens, Francis and Dufour, Sara and Van Haver, Delphi and Lopes, Graça and Rocha, António and Pinto-Leite, Rosário and Fardilha, Margarida and Colaço, Bruno}},
  issn         = {{2076-2615}},
  journal      = {{ANIMALS}},
  keywords     = {{proteomics,sperm surface proteome,sex-specific membrane proteins,sperm sexing,P-TYPE ATPASES,SPERM MOTILITY,PROTEINS,IDENTIFICATION,FERTILIZATION,ATP11C,EXPRESSION,BASIGIN,MOUSE,DAIRY}},
  language     = {{eng}},
  number       = {{4}},
  pages        = {{24}},
  title        = {{The surface proteome of bovine unsexed and sexed spermatozoa}},
  url          = {{http://doi.org/10.3390/ani15040484}},
  volume       = {{15}},
  year         = {{2025}},
}

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