Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages

Sperm cryopreservation by ultra-rapid cooling based on dropping small volumes of sperm suspension directly into liquid nitrogen, has been successful in some wild ruminant species, including the Iberian ibex (Capra pyrenaica). In ultra-rapid cooling, the contents of these droplets are expected to ent...

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Hlavní autor: Bóveda, Paula (author)
Další autoři: Toledano-Díaz, Adolfo (author), Castaño, Cristina (author), Esteso, Milagros Cristina (author), López-Sebastián, Antonio (author), Rizos, Dimitrios (author), Bielli, Alejandro (author), Ungerfeld, Rodolfo (author), Santiago-Moreno, Julián (author)
Médium: article
Jazyk:angličtina
Vydáno: 2020
Témata:
On-line přístup:https://hdl.handle.net/20.500.12008/51231
https://doi.org/10.1371/journal.pone.0227946
Tagy: Přidat tag
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author Bóveda, Paula
author2 Toledano-Díaz, Adolfo
Castaño, Cristina
Esteso, Milagros Cristina
López-Sebastián, Antonio
Rizos, Dimitrios
Bielli, Alejandro
Ungerfeld, Rodolfo
Santiago-Moreno, Julián
author2_role author
author
author
author
author
author
author
author
author_browse Bielli, Alejandro
Bóveda, Paula
Castaño, Cristina
Esteso, Milagros Cristina
López-Sebastián, Antonio
Rizos, Dimitrios
Santiago-Moreno, Julián
Toledano-Díaz, Adolfo
Ungerfeld, Rodolfo
author_facet Bóveda, Paula
Toledano-Díaz, Adolfo
Castaño, Cristina
Esteso, Milagros Cristina
López-Sebastián, Antonio
Rizos, Dimitrios
Bielli, Alejandro
Ungerfeld, Rodolfo
Santiago-Moreno, Julián
author_role author
collection COLIBRI
dc.contributor.none.fl_str_mv Bóveda Paula, INIA (Madrid, España). Departamento de Reproducción Animal
Toledano-Díaz Adolfo, INIA (Madrid, España). Departamento de Reproducción Animal
Castaño Cristina, INIA (Madrid, España). Departamento de Reproducción Animal
Esteso Milagros Cristina, INIA (Madrid, España). Departamento de Reproducción Animal
López-Sebastián Antonio, INIA (Madrid, España). Departamento de Reproducción Animal
Rizos Dimitrios, INIA (Madrid, España). Departamento de Reproducción Animal
Bielli Alejandro, Universidad de la República (Uruguay). Facultad de Veterinaria. Departamento de Morfología y Desarrollo
Ungerfeld Rodolfo, Universidad de la República (Uruguay). Facultad de Veterinaria. Departamento de Fisiología
Santiago-Moreno Julián, INIA (Madrid, España). Departamento de Reproducción Animal
dc.creator.none.fl_str_mv Bóveda, Paula
Toledano-Díaz, Adolfo
Castaño, Cristina
Esteso, Milagros Cristina
López-Sebastián, Antonio
Rizos, Dimitrios
Bielli, Alejandro
Ungerfeld, Rodolfo
Santiago-Moreno, Julián
dc.date.none.fl_str_mv 2020
2025-08-22T13:00:35Z
2025-08-22T13:00:35Z
dc.format.none.fl_str_mv 18 h
application/pdf
dc.identifier.none.fl_str_mv Bóveda, P, Toledano-Díaz, A, Castaño, C, Esteso, M, López-Sebastián, A, Rizos, D, Bielli, A, Ungerfeld, R y Santiago-Moreno, J. Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages. PLoS ONE. [en línea] 2020, 15(1): e0227946, 1-18
https://hdl.handle.net/20.500.12008/51231
https://doi.org/10.1371/journal.pone.0227946
dc.language.none.fl_str_mv en
eng
dc.relation.none.fl_str_mv PLoS ONE, 2020, 15(1): e0227946, 1-18
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
Licencia Creative Commons Atribución (CC - By 4.0)
dc.source.none.fl_str_mv reponame:COLIBRI
instname:Universidad de la República
instacron:Universidad de la República
dc.subject.none.fl_str_mv CABRAS
SEMEN
CRIOPRESERVACION
dc.title.none.fl_str_mv Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
dc.type.none.fl_str_mv Artículo
info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
description Sperm cryopreservation by ultra-rapid cooling based on dropping small volumes of sperm suspension directly into liquid nitrogen, has been successful in some wild ruminant species, including the Iberian ibex (Capra pyrenaica). In ultra-rapid cooling, the contents of these droplets are expected to enter a stable, glass-like state, but to the best of our knowledge no information exists regarding the presence or absence of ice formation in the extracellular milieu when using this technique. Different modifications to the extracellular milieu likely inflict different types of damage on the plasmalemma, the acrosome and mitochondrial membranes. The aims of the present work were: 1) to examine the physical state of the extracellular milieu after cryopreservation at slow and ultra-rapid cooling rates—and thus determine whether ultra-rapid cooling vitrifies the extracellular milieu; and 2) to compare, using conventional sperm analysis techniques and scanning and transmission electron microscopy, the damage to sperm caused by these two methods. Sperm samples were obtained by the transrectal ultrasound-guided massage method (TUMASG) from anesthetized Iberian ibexes, and cryopreserved using slow and ultra-rapid cooling techniques. Sperm motility (22.95 ± 3.22% vs 4.42 ± 0.86%), viability (25.64 ± 3.71% vs 12.8 ± 2.50%), acrosome integrity (41.45± 3.73% vs 27.00 ± 1.84%) and mitochondrial membrane integrity (16.52 ± 3.75% vs 4.00 ± 0.65%) were better after slow cooling (P<0.001) than after ultrarapid technique. Cryo-scanning electron microscopy (Cryo-SEM) suggested that the vitrified state was not achieved by ultra-rapid cooling, and that the ice crystals formed were smaller and had more stretchmarks (P<0.001) than after slow cooling. Scanning electron microscopy revealed no differences in the types of damage caused by the examined techniques, although transmission electron microscopy showed the damage to the plasmalemma and mitochondrial membrane to be worse after ultra-rapid cooling. In conclusion ultra-rapid cooling provoked more membrane damage than slow cooling, perhaps due to the extracellular ice crystals formed.
eu_rights_str_mv openAccess
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identifier_str_mv Bóveda, P, Toledano-Díaz, A, Castaño, C, Esteso, M, López-Sebastián, A, Rizos, D, Bielli, A, Ungerfeld, R y Santiago-Moreno, J. Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages. PLoS ONE. [en línea] 2020, 15(1): e0227946, 1-18
instacron_str Universidad de la República
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publishDate 2020
publishDateSort 2020
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rights_invalid_str_mv Licencia Creative Commons Atribución (CC - By 4.0)
spelling Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damagesBóveda, PaulaToledano-Díaz, AdolfoCastaño, CristinaEsteso, Milagros CristinaLópez-Sebastián, AntonioRizos, DimitriosBielli, AlejandroUngerfeld, RodolfoSantiago-Moreno, JuliánCABRASSEMENCRIOPRESERVACIONSperm cryopreservation by ultra-rapid cooling based on dropping small volumes of sperm suspension directly into liquid nitrogen, has been successful in some wild ruminant species, including the Iberian ibex (Capra pyrenaica). In ultra-rapid cooling, the contents of these droplets are expected to enter a stable, glass-like state, but to the best of our knowledge no information exists regarding the presence or absence of ice formation in the extracellular milieu when using this technique. Different modifications to the extracellular milieu likely inflict different types of damage on the plasmalemma, the acrosome and mitochondrial membranes. The aims of the present work were: 1) to examine the physical state of the extracellular milieu after cryopreservation at slow and ultra-rapid cooling rates—and thus determine whether ultra-rapid cooling vitrifies the extracellular milieu; and 2) to compare, using conventional sperm analysis techniques and scanning and transmission electron microscopy, the damage to sperm caused by these two methods. Sperm samples were obtained by the transrectal ultrasound-guided massage method (TUMASG) from anesthetized Iberian ibexes, and cryopreserved using slow and ultra-rapid cooling techniques. Sperm motility (22.95 ± 3.22% vs 4.42 ± 0.86%), viability (25.64 ± 3.71% vs 12.8 ± 2.50%), acrosome integrity (41.45± 3.73% vs 27.00 ± 1.84%) and mitochondrial membrane integrity (16.52 ± 3.75% vs 4.00 ± 0.65%) were better after slow cooling (P<0.001) than after ultrarapid technique. Cryo-scanning electron microscopy (Cryo-SEM) suggested that the vitrified state was not achieved by ultra-rapid cooling, and that the ice crystals formed were smaller and had more stretchmarks (P<0.001) than after slow cooling. Scanning electron microscopy revealed no differences in the types of damage caused by the examined techniques, although transmission electron microscopy showed the damage to the plasmalemma and mitochondrial membrane to be worse after ultra-rapid cooling. In conclusion ultra-rapid cooling provoked more membrane damage than slow cooling, perhaps due to the extracellular ice crystals formed.Bóveda Paula, INIA (Madrid, España). Departamento de Reproducción AnimalToledano-Díaz Adolfo, INIA (Madrid, España). Departamento de Reproducción AnimalCastaño Cristina, INIA (Madrid, España). Departamento de Reproducción AnimalEsteso Milagros Cristina, INIA (Madrid, España). Departamento de Reproducción AnimalLópez-Sebastián Antonio, INIA (Madrid, España). Departamento de Reproducción AnimalRizos Dimitrios, INIA (Madrid, España). Departamento de Reproducción AnimalBielli Alejandro, Universidad de la República (Uruguay). Facultad de Veterinaria. Departamento de Morfología y DesarrolloUngerfeld Rodolfo, Universidad de la República (Uruguay). Facultad de Veterinaria. Departamento de FisiologíaSantiago-Moreno Julián, INIA (Madrid, España). Departamento de Reproducción Animal2025-08-22T13:00:35Z2025-08-22T13:00:35Z2020Artículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion18 happlication/pdfBóveda, P, Toledano-Díaz, A, Castaño, C, Esteso, M, López-Sebastián, A, Rizos, D, Bielli, A, Ungerfeld, R y Santiago-Moreno, J. Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages. PLoS ONE. [en línea] 2020, 15(1): e0227946, 1-18https://hdl.handle.net/20.500.12008/51231https://doi.org/10.1371/journal.pone.0227946reponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaenengPLoS ONE, 2020, 15(1): e0227946, 1-18Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución (CC - By 4.0)oai:colibri.udelar.edu.uy:20.500.12008/512312026-04-14T10:35:07Z
spellingShingle Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
Bóveda, Paula
CABRAS
SEMEN
CRIOPRESERVACION
status_str publishedVersion
title Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
title_full Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
title_fullStr Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
title_full_unstemmed Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
title_short Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
title_sort Ultra-rapid cooling of ibex sperm by spheres method does not induce a vitreous extracellular state and increases the membrane damages
topic CABRAS
SEMEN
CRIOPRESERVACION
url https://hdl.handle.net/20.500.12008/51231
https://doi.org/10.1371/journal.pone.0227946