Comparison of European eel sperm cryopreservation protocols with standardization as a target

[EN] The critical situation of the European eel (Anguilla anguilla) has urged the development of sperm cryopreservation protocols for reproduction in captivity and cryobanking. In the last years, two research groups have developed their own protocols in Spain and Hungary with positive results, but d...

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Autores: Herranz-Jusdado, Juan Germán, MORINI, MARINA, Rozenfeld, Christoffer, Kása, E., Kollár, T., Depincé, A., Labbé, C., Horváth, Á., Gallego Albiach, Victor|||0000-0002-9733-0625, Pérez Igualada, Luz María|||0000-0002-6678-7446, Asturiano, Juan F.|||0000-0002-6441-5294
Tipo de recurso: artículo
Fecha de publicación:2019
País:España
Institución:Universitat Politècnica de València (UPV)
Repositorio:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Idioma:inglés
OAI Identifier:oai:riunet.upv.es:10251/120142
Acceso en línea:https://riunet.upv.es/handle/10251/120142
Access Level:acceso abierto
Palabra clave:Anguilla anguilla
Methanol
DMSO
DNA methylation
Epigenetics
ZOOLOGIA
PRODUCCION ANIMAL
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oai_identifier_str oai:riunet.upv.es:10251/120142
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Comparison of European eel sperm cryopreservation protocols with standardization as a target
title Comparison of European eel sperm cryopreservation protocols with standardization as a target
spellingShingle Comparison of European eel sperm cryopreservation protocols with standardization as a target
Herranz-Jusdado, Juan Germán
Anguilla anguilla
Methanol
DMSO
DNA methylation
Epigenetics
ZOOLOGIA
PRODUCCION ANIMAL
title_short Comparison of European eel sperm cryopreservation protocols with standardization as a target
title_full Comparison of European eel sperm cryopreservation protocols with standardization as a target
title_fullStr Comparison of European eel sperm cryopreservation protocols with standardization as a target
title_full_unstemmed Comparison of European eel sperm cryopreservation protocols with standardization as a target
title_sort Comparison of European eel sperm cryopreservation protocols with standardization as a target
dc.creator.none.fl_str_mv Herranz-Jusdado, Juan Germán
MORINI, MARINA
Rozenfeld, Christoffer
Kása, E.
Kollár, T.
Depincé, A.
Labbé, C.
Horváth, Á.
Gallego Albiach, Victor|||0000-0002-9733-0625
Pérez Igualada, Luz María|||0000-0002-6678-7446
Asturiano, Juan F.|||0000-0002-6441-5294
author Herranz-Jusdado, Juan Germán
author_facet Herranz-Jusdado, Juan Germán
MORINI, MARINA
Rozenfeld, Christoffer
Kása, E.
Kollár, T.
Depincé, A.
Labbé, C.
Horváth, Á.
Gallego Albiach, Victor|||0000-0002-9733-0625
Pérez Igualada, Luz María|||0000-0002-6678-7446
Asturiano, Juan F.|||0000-0002-6441-5294
author_role author
author2 MORINI, MARINA
Rozenfeld, Christoffer
Kása, E.
Kollár, T.
Depincé, A.
Labbé, C.
Horváth, Á.
Gallego Albiach, Victor|||0000-0002-9733-0625
Pérez Igualada, Luz María|||0000-0002-6678-7446
Asturiano, Juan F.|||0000-0002-6441-5294
author2_role author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Departamento de Ciencia Animal
Instituto Universitario de Ciencia y Tecnología Animal
Escuela Técnica Superior de Ingeniería Agronómica y del Medio Natural
European Commission
Repositorio Institucional de la Universitat Politècnica de València Riunet
dc.subject.none.fl_str_mv Anguilla anguilla
Methanol
DMSO
DNA methylation
Epigenetics
ZOOLOGIA
PRODUCCION ANIMAL
topic Anguilla anguilla
Methanol
DMSO
DNA methylation
Epigenetics
ZOOLOGIA
PRODUCCION ANIMAL
description [EN] The critical situation of the European eel (Anguilla anguilla) has urged the development of sperm cryopreservation protocols for reproduction in captivity and cryobanking. In the last years, two research groups have developed their own protocols in Spain and Hungary with positive results, but difficult to compare. Here, a series of experiments were conducted to test the quality of thawed sperm after using both protocols, determining which of them produce the best results and aiming for standardization. The quality of thawed sperm was assessed by studying the motility and kinetic values of thawed sperm from both cryopreservation protocols using a computer-assisted sperm analysis (CASA-Mot) system. In addition, a viability analysis was performed using flow cytometry to test if the cryoprotectants or the freezing-thawing process led to a reduction in spermatozoa survival. Furthermore, since during cryopreservation the sperm was treated with methylated cryoprotectants (DMSO or methanol) that may induce epigenetic changes in the sperm DNA (cytosine methylation) and could affect the offspring, we conducted a luminometric methylation assay (LUMA) to study the DNA methylation levels induced by both protocols. In this work, all the above-mentioned parameters were analyzed in fresh and frozen-thawed sperm samples. Our results showed that thawed sperm samples from both protocols presented lower sperm motility and velocity, and lower percentage of live cells than those shown in fresh sperm samples. Furthermore, sperm samples from the methanol based protocol showed significantly higher motility, velocity and percentage of live spermatozoa than the same sperm samples treated with the DMSO based protocol. In addition, the DMSO based protocol induced a hypomethylation of sperm DNA compared to fresh samples whereas the methanol based protocol did not alter sperm DNA methylation level. Our results indicate that the methanol based protocol is a more suitable protocol that preserves better the motility and genetic qualities of the European eel sperm.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019-01-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://riunet.upv.es/handle/10251/120142
url https://riunet.upv.es/handle/10251/120142
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv European Commission https://doi.org/10.13039/501100000780 H2020 642893 Improved production strategies for endangered freshwater species.
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Reconocimiento (by)
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
instname:Universitat Politècnica de València (UPV)
instname_str Universitat Politècnica de València (UPV)
reponame_str RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
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spelling Comparison of European eel sperm cryopreservation protocols with standardization as a targetHerranz-Jusdado, Juan GermánMORINI, MARINARozenfeld, ChristofferKása, E.Kollár, T.Depincé, A.Labbé, C.Horváth, Á.Gallego Albiach, Victor|||0000-0002-9733-0625Pérez Igualada, Luz María|||0000-0002-6678-7446Asturiano, Juan F.|||0000-0002-6441-5294Anguilla anguillaMethanolDMSODNA methylationEpigeneticsZOOLOGIAPRODUCCION ANIMAL[EN] The critical situation of the European eel (Anguilla anguilla) has urged the development of sperm cryopreservation protocols for reproduction in captivity and cryobanking. In the last years, two research groups have developed their own protocols in Spain and Hungary with positive results, but difficult to compare. Here, a series of experiments were conducted to test the quality of thawed sperm after using both protocols, determining which of them produce the best results and aiming for standardization. The quality of thawed sperm was assessed by studying the motility and kinetic values of thawed sperm from both cryopreservation protocols using a computer-assisted sperm analysis (CASA-Mot) system. In addition, a viability analysis was performed using flow cytometry to test if the cryoprotectants or the freezing-thawing process led to a reduction in spermatozoa survival. Furthermore, since during cryopreservation the sperm was treated with methylated cryoprotectants (DMSO or methanol) that may induce epigenetic changes in the sperm DNA (cytosine methylation) and could affect the offspring, we conducted a luminometric methylation assay (LUMA) to study the DNA methylation levels induced by both protocols. In this work, all the above-mentioned parameters were analyzed in fresh and frozen-thawed sperm samples. Our results showed that thawed sperm samples from both protocols presented lower sperm motility and velocity, and lower percentage of live cells than those shown in fresh sperm samples. Furthermore, sperm samples from the methanol based protocol showed significantly higher motility, velocity and percentage of live spermatozoa than the same sperm samples treated with the DMSO based protocol. In addition, the DMSO based protocol induced a hypomethylation of sperm DNA compared to fresh samples whereas the methanol based protocol did not alter sperm DNA methylation level. Our results indicate that the methanol based protocol is a more suitable protocol that preserves better the motility and genetic qualities of the European eel sperm.Funded by the European Union's Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement N 642893 (IMPRESS), including the JGHJ and CR predoctoral contracts, the French CRB Anim project "Investissements d'avenir", ANR-11-INBS-0003, as well as the Hungarian EFOP 3.6.3.-VEKOP 16.-2017-00008 project, and by the Higher Education Institutional Excellence Program (1783-3/2018/FEKUTSRAT) awarded by the Ministry of Human Capacities within the framework of water related researches of Szent Istvan University. MM, EK, TK and AH were granted with Short Term Scientific Missions by the COST Office (COST Action FA1205: AQUAGAMETE).ElsevierDepartamento de Ciencia AnimalInstituto Universitario de Ciencia y Tecnología AnimalEscuela Técnica Superior de Ingeniería Agronómica y del Medio NaturalEuropean CommissionRepositorio Institucional de la Universitat Politècnica de València Riunet20192019-01-01journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfapplication/pdfhttps://riunet.upv.es/handle/10251/120142reponame:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valénciainstname:Universitat Politècnica de València (UPV)InglésengEuropean Commission https://doi.org/10.13039/501100000780 H2020 642893 Improved production strategies for endangered freshwater species.open accesshttp://purl.org/coar/access_right/c_abf2Reconocimiento (by)http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:riunet.upv.es:10251/1201422026-06-13T07:49:27Z
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