Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoa
Reproduction is fundamental to breeding programs aimed at increasing productivity in swine industry. However, the application of in vitro embryo production in this species is limited because of the polyspermy. Therefore, characterizing proteins involved in sperm-oocyte binding such as IZUMO1 becomes...
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Frontiers Media S.A.
2025-05-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2025.1576881/full |
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| author | Miranda Hernández-Falcó Paula Sáez-Espinosa Andrea López-Botella Laura Robles-Gómez Francisco Alberto García-Vázquez Francisco Alberto García-Vázquez Maria José Izquierdo-Rico Maria José Izquierdo-Rico Pedro José Llamas-López María José Gómez-Torres María José Gómez-Torres |
| author_facet | Miranda Hernández-Falcó Paula Sáez-Espinosa Andrea López-Botella Laura Robles-Gómez Francisco Alberto García-Vázquez Francisco Alberto García-Vázquez Maria José Izquierdo-Rico Maria José Izquierdo-Rico Pedro José Llamas-López María José Gómez-Torres María José Gómez-Torres |
| author_sort | Miranda Hernández-Falcó |
| collection | DOAJ |
| description | Reproduction is fundamental to breeding programs aimed at increasing productivity in swine industry. However, the application of in vitro embryo production in this species is limited because of the polyspermy. Therefore, characterizing proteins involved in sperm-oocyte binding such as IZUMO1 becomes essential. This study aimed to characterize porcine IZUMO1 protein under three different physiological states: sperm-rich fraction (SRF), 1-h capacitated sperm selected by swim-up (CS), and induced acrosome reaction in 1-h capacitated sperm (ARS). The immunolocalization of IZUMO1 and acrosome status of fifteen fertile boars was assessed by confocal microscopy. Additionally, six males were subjected to a more detailed examination via quantitative proteomic analysis by LC–MS/MS. Fluorescence results revealed four distinct IZUMO1 distribution patterns: pattern 1 (P1) characterized by speckled staining in the pre-equatorial subdomain and postacrosomal domain, pattern 2 (P2) displaying strong apical ridge staining with speckled staining in the pre-equatorial subdomain and postacrosomal domain, pattern 3 (P3) exhibiting speckled staining in the postacrosomal domain, and pattern 4 (P4) without labelling. In the SRF sperm, IZUMO1 was predominantly distributed between staining patterns P1 and P2 (∼50%). As a result of the capacitation, there was a significant decrease in P1. Conversely, in ARS, IZUMO1 was dominantly distributed in P3 51.55% and P4 24.25%. The quantitative study of the IZUMO1 protein supported these findings. With those results and compared with our previous work in human, here we propose a working model of IZUMO1 migration dependent on the morphology and subdomains of the sperm head. |
| format | Article |
| id | doaj-art-1d2693ec81c04445a8029263a6694ea5 |
| institution | DOAJ |
| issn | 2296-634X |
| language | English |
| publishDate | 2025-05-01 |
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| spelling | doaj-art-1d2693ec81c04445a8029263a6694ea52025-08-20T03:08:16ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2025-05-011310.3389/fcell.2025.15768811576881Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoaMiranda Hernández-Falcó0Paula Sáez-Espinosa1Andrea López-Botella2Laura Robles-Gómez3Francisco Alberto García-Vázquez4Francisco Alberto García-Vázquez5Maria José Izquierdo-Rico6Maria José Izquierdo-Rico7Pedro José Llamas-López8María José Gómez-Torres9María José Gómez-Torres10Department of Biotechnology, University of Alicante, Alicante, SpainDepartment of Biotechnology, University of Alicante, Alicante, SpainDepartment of Biotechnology, University of Alicante, Alicante, SpainDepartment of Biotechnology, University of Alicante, Alicante, SpainDepartamento de Fisiología, Facultad de Veterinaria, Universidad de Murcia, Murcia, SpainInstituto Murciano de Investigación Biosanitaria (IMIB-Pascual Parrilla), Murcia, SpainInstituto Murciano de Investigación Biosanitaria (IMIB-Pascual Parrilla), Murcia, SpainDepartamento de Biología Celular e Histología, Facultad de Medicina, Universidad de Murcia, Murcia, SpainDepartment of Agri-Food Technology, University of Miguel Hernández, Elche, SpainDepartment of Biotechnology, University of Alicante, Alicante, SpainHuman Fertility Cathedra, University of Alicante, Alicante, SpainReproduction is fundamental to breeding programs aimed at increasing productivity in swine industry. However, the application of in vitro embryo production in this species is limited because of the polyspermy. Therefore, characterizing proteins involved in sperm-oocyte binding such as IZUMO1 becomes essential. This study aimed to characterize porcine IZUMO1 protein under three different physiological states: sperm-rich fraction (SRF), 1-h capacitated sperm selected by swim-up (CS), and induced acrosome reaction in 1-h capacitated sperm (ARS). The immunolocalization of IZUMO1 and acrosome status of fifteen fertile boars was assessed by confocal microscopy. Additionally, six males were subjected to a more detailed examination via quantitative proteomic analysis by LC–MS/MS. Fluorescence results revealed four distinct IZUMO1 distribution patterns: pattern 1 (P1) characterized by speckled staining in the pre-equatorial subdomain and postacrosomal domain, pattern 2 (P2) displaying strong apical ridge staining with speckled staining in the pre-equatorial subdomain and postacrosomal domain, pattern 3 (P3) exhibiting speckled staining in the postacrosomal domain, and pattern 4 (P4) without labelling. In the SRF sperm, IZUMO1 was predominantly distributed between staining patterns P1 and P2 (∼50%). As a result of the capacitation, there was a significant decrease in P1. Conversely, in ARS, IZUMO1 was dominantly distributed in P3 51.55% and P4 24.25%. The quantitative study of the IZUMO1 protein supported these findings. With those results and compared with our previous work in human, here we propose a working model of IZUMO1 migration dependent on the morphology and subdomains of the sperm head.https://www.frontiersin.org/articles/10.3389/fcell.2025.1576881/fullacrosome reactionboarcapacitationimmunofluorescenceIZUMO1proteomics |
| spellingShingle | Miranda Hernández-Falcó Paula Sáez-Espinosa Andrea López-Botella Laura Robles-Gómez Francisco Alberto García-Vázquez Francisco Alberto García-Vázquez Maria José Izquierdo-Rico Maria José Izquierdo-Rico Pedro José Llamas-López María José Gómez-Torres María José Gómez-Torres Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoa Frontiers in Cell and Developmental Biology acrosome reaction boar capacitation immunofluorescence IZUMO1 proteomics |
| title | Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoa |
| title_full | Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoa |
| title_fullStr | Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoa |
| title_full_unstemmed | Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoa |
| title_short | Immunolocalization and proteomic analyses of IZUMO1 in porcine spermatozoa |
| title_sort | immunolocalization and proteomic analyses of izumo1 in porcine spermatozoa |
| topic | acrosome reaction boar capacitation immunofluorescence IZUMO1 proteomics |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2025.1576881/full |
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