Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it?
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1371/journal.pone.0160988 http://hdl.handle.net/11449/173428 |
Resumo: | Boar semen cryopreservation remains a challenge due to the extension of cold shock damage. Thus, many alternatives have emerged to improve the quality of frozen-thawed boar sperm. Although the use of seminal plasma arising from boar sperm-rich fraction (SP-SRF) has shown good efficacy; however, the majority of actual sperm evaluation techniques include a single or dual sperm parameter analysis, which overrates the real sperm viability. Within this context, this work was performed to introduce a sperm flow cytometry fourfold stain technique for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential. We then used the sperm flow cytometry fourfold stain technique to study the effect of SP-SRF on frozen-thawed boar sperm and further evaluated the effect of this treatment on sperm movement, tyrosine phosphorylation and fertility rate (FR). The sperm fourfold stain technique is accurate (R2 = 0.9356, p > 0.01) for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential (IPIAH cells). Centrifugation pre-cryopreservation was not deleterious (p > 0.05) for any analyzed variables. Addition of SP-SRF after cryopreservation was able to improve total and progressive motility (p < 0.05) when boar semen was cryopreserved without SP-SRF; however, it was not able to decrease tyrosine phosphorylation (p > 0.05) or improve IPIAH cells (p > 0.05). FR was not (p > 0.05) statistically increased by the addition of seminal plasma, though females inseminated with frozen-thawed boar semen plus SP-SRF did perform better than those inseminated with sperm lacking seminal plasma. Thus, we conclude that sperm fourfold stain can be used to simultaneously evaluate plasma and acrosomal membrane integrity and mitochondrial membrane potential, and the addition of SP-SRF at thawed boar semen cryopreserved in absence of SP-SRF improve its total and progressive motility. |
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Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it?Boar semen cryopreservation remains a challenge due to the extension of cold shock damage. Thus, many alternatives have emerged to improve the quality of frozen-thawed boar sperm. Although the use of seminal plasma arising from boar sperm-rich fraction (SP-SRF) has shown good efficacy; however, the majority of actual sperm evaluation techniques include a single or dual sperm parameter analysis, which overrates the real sperm viability. Within this context, this work was performed to introduce a sperm flow cytometry fourfold stain technique for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential. We then used the sperm flow cytometry fourfold stain technique to study the effect of SP-SRF on frozen-thawed boar sperm and further evaluated the effect of this treatment on sperm movement, tyrosine phosphorylation and fertility rate (FR). The sperm fourfold stain technique is accurate (R2 = 0.9356, p > 0.01) for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential (IPIAH cells). Centrifugation pre-cryopreservation was not deleterious (p > 0.05) for any analyzed variables. Addition of SP-SRF after cryopreservation was able to improve total and progressive motility (p < 0.05) when boar semen was cryopreserved without SP-SRF; however, it was not able to decrease tyrosine phosphorylation (p > 0.05) or improve IPIAH cells (p > 0.05). FR was not (p > 0.05) statistically increased by the addition of seminal plasma, though females inseminated with frozen-thawed boar semen plus SP-SRF did perform better than those inseminated with sperm lacking seminal plasma. Thus, we conclude that sperm fourfold stain can be used to simultaneously evaluate plasma and acrosomal membrane integrity and mitochondrial membrane potential, and the addition of SP-SRF at thawed boar semen cryopreserved in absence of SP-SRF improve its total and progressive motility.Laboratory of Andrology and Technology of Swine Embryos Department of Animal Reproduction School of Veterinary Medicine and Animal Science University of São PauloCenter of Excellence in Biotechnology of Reproduction University of la FronteraLaboratory of Swine Research Department of Animal Nutrition and Production School of Veterinary Medicine and Animal Science University of São PauloDepartment of Veterinary Medicine School of Animal Sciences and Food Engineerig University of São PauloDepartment of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science São Paulo State University (UNESP)Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science São Paulo State University (UNESP)Universidade de São Paulo (USP)University of la FronteraUniversidade Estadual Paulista (Unesp)Torres, Mariana AndradeDíaz, RommyBoguen, RodrigoMartins, Simone Maria Massami KitamuraRavagnani, Gisele MouroLeal, Diego FeitosaDe Lima Oliveira, MelissaMuro, Bruno Bracco DonatelliParra, Beatriz MartinsMeirelles, Flávio VieiraPapa, Frederico Ozanan [UNESP]Dell'Aqua, José Antônio [UNESP]Alvarenga, Marco Antônio [UNESP]De Sant'Anna Moretti, AníbalSepúlveda, NéstorDe Andrade, André Furugen Cesar2018-12-11T17:05:24Z2018-12-11T17:05:24Z2016-08-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1371/journal.pone.0160988PLoS ONE, v. 11, n. 8, 2016.1932-6203http://hdl.handle.net/11449/17342810.1371/journal.pone.01609882-s2.0-849847997342-s2.0-84984799734.pdf0473846154288947Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPLoS ONE1,164info:eu-repo/semantics/openAccess2024-09-09T14:01:08Zoai:repositorio.unesp.br:11449/173428Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462024-09-09T14:01:08Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? |
title |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? |
spellingShingle |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? Torres, Mariana Andrade |
title_short |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? |
title_full |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? |
title_fullStr |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? |
title_full_unstemmed |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? |
title_sort |
Novel flow cytometry analyses of boar sperm viability: Can the addition of whole sperm-rich fraction seminal plasma to frozen-thawed boar sperm affect it? |
author |
Torres, Mariana Andrade |
author_facet |
Torres, Mariana Andrade Díaz, Rommy Boguen, Rodrigo Martins, Simone Maria Massami Kitamura Ravagnani, Gisele Mouro Leal, Diego Feitosa De Lima Oliveira, Melissa Muro, Bruno Bracco Donatelli Parra, Beatriz Martins Meirelles, Flávio Vieira Papa, Frederico Ozanan [UNESP] Dell'Aqua, José Antônio [UNESP] Alvarenga, Marco Antônio [UNESP] De Sant'Anna Moretti, Aníbal Sepúlveda, Néstor De Andrade, André Furugen Cesar |
author_role |
author |
author2 |
Díaz, Rommy Boguen, Rodrigo Martins, Simone Maria Massami Kitamura Ravagnani, Gisele Mouro Leal, Diego Feitosa De Lima Oliveira, Melissa Muro, Bruno Bracco Donatelli Parra, Beatriz Martins Meirelles, Flávio Vieira Papa, Frederico Ozanan [UNESP] Dell'Aqua, José Antônio [UNESP] Alvarenga, Marco Antônio [UNESP] De Sant'Anna Moretti, Aníbal Sepúlveda, Néstor De Andrade, André Furugen Cesar |
author2_role |
author author author author author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) University of la Frontera Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Torres, Mariana Andrade Díaz, Rommy Boguen, Rodrigo Martins, Simone Maria Massami Kitamura Ravagnani, Gisele Mouro Leal, Diego Feitosa De Lima Oliveira, Melissa Muro, Bruno Bracco Donatelli Parra, Beatriz Martins Meirelles, Flávio Vieira Papa, Frederico Ozanan [UNESP] Dell'Aqua, José Antônio [UNESP] Alvarenga, Marco Antônio [UNESP] De Sant'Anna Moretti, Aníbal Sepúlveda, Néstor De Andrade, André Furugen Cesar |
description |
Boar semen cryopreservation remains a challenge due to the extension of cold shock damage. Thus, many alternatives have emerged to improve the quality of frozen-thawed boar sperm. Although the use of seminal plasma arising from boar sperm-rich fraction (SP-SRF) has shown good efficacy; however, the majority of actual sperm evaluation techniques include a single or dual sperm parameter analysis, which overrates the real sperm viability. Within this context, this work was performed to introduce a sperm flow cytometry fourfold stain technique for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential. We then used the sperm flow cytometry fourfold stain technique to study the effect of SP-SRF on frozen-thawed boar sperm and further evaluated the effect of this treatment on sperm movement, tyrosine phosphorylation and fertility rate (FR). The sperm fourfold stain technique is accurate (R2 = 0.9356, p > 0.01) for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential (IPIAH cells). Centrifugation pre-cryopreservation was not deleterious (p > 0.05) for any analyzed variables. Addition of SP-SRF after cryopreservation was able to improve total and progressive motility (p < 0.05) when boar semen was cryopreserved without SP-SRF; however, it was not able to decrease tyrosine phosphorylation (p > 0.05) or improve IPIAH cells (p > 0.05). FR was not (p > 0.05) statistically increased by the addition of seminal plasma, though females inseminated with frozen-thawed boar semen plus SP-SRF did perform better than those inseminated with sperm lacking seminal plasma. Thus, we conclude that sperm fourfold stain can be used to simultaneously evaluate plasma and acrosomal membrane integrity and mitochondrial membrane potential, and the addition of SP-SRF at thawed boar semen cryopreserved in absence of SP-SRF improve its total and progressive motility. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-08-01 2018-12-11T17:05:24Z 2018-12-11T17:05:24Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1371/journal.pone.0160988 PLoS ONE, v. 11, n. 8, 2016. 1932-6203 http://hdl.handle.net/11449/173428 10.1371/journal.pone.0160988 2-s2.0-84984799734 2-s2.0-84984799734.pdf 0473846154288947 |
url |
http://dx.doi.org/10.1371/journal.pone.0160988 http://hdl.handle.net/11449/173428 |
identifier_str_mv |
PLoS ONE, v. 11, n. 8, 2016. 1932-6203 10.1371/journal.pone.0160988 2-s2.0-84984799734 2-s2.0-84984799734.pdf 0473846154288947 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
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PLoS ONE 1,164 |
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info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
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Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
repositoriounesp@unesp.br |
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1810021321685336064 |