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Comparative analysis of boar seminal plasma proteome from different freezability ejaculates and identification of Fibronectin 1 as sperm freezability marker

Variation in boar sperm freezability (i.e. capacity to withstand cryopreservation) between ejaculates is a limitation largely reported in the literature. Prediction of sperm freezability and classification of boar ejaculates into good (GFEs) and poor freezability ejaculates (PFEs) before cryopreservation takes place may increase the use of frozen-thawed spermatozoa. While markers of boar sperm freezability have been found from sperm cell extracts, little attention has been paid to seminal plasma. On this basis, the present study compared the fresh seminal plasma proteome of 9 GFEs and 9 PFEs through two-dimensional difference gel electrophoresis (2D-DIGE) and liquid chromatography mass spectrometry (LC-MS/MS). The ejaculates were previously classified as GFE or PFE upon their sperm viability and progressive motility assessments at 30 and 240 min post thawing. From a total of 51 spots, four were found to significantly (p < 0.05) differ between GFEs and PFEs, and two were identified as fibronectin-1 (FN1) and glutathione peroxidase 5 (GPX5). These two potential markers were further studied by western blot and correlation analysis between protein relative abundances in fresh seminal plasma and regression factors from principal component analyses (PCA) run using post-thawing sperm quality parameters. Results confirmed that FN1 is a reliable marker of boar sperm freezability, because GFEs presented significantly (p < 0.05) higher FN1-amounts than PFEs and FN1 was found to be correlated with the first PCA component at 240 min post thawing. In contrast, GPX5 was not validated as a boar sperm freezability marker. We can thus conclude that levels of FN1 in fresh seminal plasma from boar semen may be used as a sperm freezability marker, thereby facilitating the use of frozen-thawed boar spermatozoa

This research was supported by Grant Id. RZ 2011-00001-00-00, Spanish Ministry of Science and Innovation, and by a PhD studentship granted to Ingrid Vilagran from University of Girona. Proteomic analyses were carried out at the Proteomic Unit of the Scientific and Technological Centres, University of Barcelona (CCiTUB), a member of the ProteoRed network

© Andrology, 2015, vol. 3, núm. 2, p. 345-356

American Society of Andrology and European Academy of Andrology

Author: Vilagran, Ingrid
Yeste Oliveras, Marc
Sancho Badell, Sílvia
Castillo Martín, Miriam
Oliva, Rafael
Bonet, Sergi
Date: 2015
Abstract: Variation in boar sperm freezability (i.e. capacity to withstand cryopreservation) between ejaculates is a limitation largely reported in the literature. Prediction of sperm freezability and classification of boar ejaculates into good (GFEs) and poor freezability ejaculates (PFEs) before cryopreservation takes place may increase the use of frozen-thawed spermatozoa. While markers of boar sperm freezability have been found from sperm cell extracts, little attention has been paid to seminal plasma. On this basis, the present study compared the fresh seminal plasma proteome of 9 GFEs and 9 PFEs through two-dimensional difference gel electrophoresis (2D-DIGE) and liquid chromatography mass spectrometry (LC-MS/MS). The ejaculates were previously classified as GFE or PFE upon their sperm viability and progressive motility assessments at 30 and 240 min post thawing. From a total of 51 spots, four were found to significantly (p < 0.05) differ between GFEs and PFEs, and two were identified as fibronectin-1 (FN1) and glutathione peroxidase 5 (GPX5). These two potential markers were further studied by western blot and correlation analysis between protein relative abundances in fresh seminal plasma and regression factors from principal component analyses (PCA) run using post-thawing sperm quality parameters. Results confirmed that FN1 is a reliable marker of boar sperm freezability, because GFEs presented significantly (p < 0.05) higher FN1-amounts than PFEs and FN1 was found to be correlated with the first PCA component at 240 min post thawing. In contrast, GPX5 was not validated as a boar sperm freezability marker. We can thus conclude that levels of FN1 in fresh seminal plasma from boar semen may be used as a sperm freezability marker, thereby facilitating the use of frozen-thawed boar spermatozoa
This research was supported by Grant Id. RZ 2011-00001-00-00, Spanish Ministry of Science and Innovation, and by a PhD studentship granted to Ingrid Vilagran from University of Girona. Proteomic analyses were carried out at the Proteomic Unit of the Scientific and Technological Centres, University of Barcelona (CCiTUB), a member of the ProteoRed network
Format: application/pdf
ISSN: 2047-2919 (versió paper)
2047-2927 (versió electrònica)
Document access: http://hdl.handle.net/10256/12393
Language: eng
Publisher: American Society of Andrology and European Academy of Andrology
Collection: MICINN/PN 2011-2014/RZ 2011-00001-00-00
Reproducció digital del document publicat a: http://dx.doi.org/10.1111/andr.12009
Articles publicats (D-B)
Is part of: © Andrology, 2015, vol. 3, núm. 2, p. 345-356
Rights: Tots els drets reservats
Subject: Semen -- Crioconservació
semen -- Cryopreservation
Porcs -- Espermatozoides
Swine -- Spermatozoa
Title: Comparative analysis of boar seminal plasma proteome from different freezability ejaculates and identification of Fibronectin 1 as sperm freezability marker
Type: info:eu-repo/semantics/article
Repository: DUGiDocs

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