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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| Médium: | article |
| Jazyk: | angličtina |
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2020
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| On-line přístup: | https://hdl.handle.net/20.500.12008/51231 https://doi.org/10.1371/journal.pone.0227946 |
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| _version_ | 1868890215962116096 |
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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 |
| format | article |
| id | anni_e22d1b0e4cf8bb9d4127969d2a9871e8 |
| 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 |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | anni |
| network_name_str | oai-lr-anni |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/51231 |
| publishDate | 2020 |
| publishDateSort | 2020 |
| reponame_str | COLIBRI |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| 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 |