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Cryoprotectants synergy improve zebrafish sperm cryopreservation and offspring skeletogenesis

2019-12Journal article Restricted access
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dc.contributor.authorDiogo, Patricia
dc.contributor.authorMartins, Gil
dc.contributor.authorNogueira, Rita
dc.contributor.authorMarreiros, Ana
dc.contributor.authorGavaia, Paulo
dc.contributor.authorCabrita, Elsa
dc.date.accessioned2020-07-24T10:50:48Z
dc.date.available2020-07-24T10:50:48Z
dc.date.issued2019-12
dc.description.abstractThe synergy obtained by the combination of cryoprotectants is a successful strategy that can be beneficial on the optimization of zebrafish sperm cryopreservation. Recently, a protocol was established for this species using an electric ultrafreezer (-150 degrees C) performing cooling rate (-66 degrees C/min) and storage within one step. The ultimate objective of sperm cryopreservation is to generate healthy offspring. Therefore, the objective of this study was to select the most adequate cryoprotectant combination, for the previously established protocol, that generate high quality offspring with normal skeletogenesis. Among the permeating cryoprotectant concentrations studied 12.5% and 15% of N,N-dimethylformamide (DMF) yielded high post-thaw sperm quality and hatching rates. For these two concentrations, the presence of bovine serum albumin (10 mg/mL), egg yolk (10%), glycine (30 mM) and bicine (50 mM) was evaluated for post-thaw sperm motility, viability, in vitro fertilization success and offspring skeletal development (30 days post fertilization). Higher concentration of permeating cryoprotectant (15%) decreased the incidence of deformed arches and severe skeletal malformations, which suggests higher capacity to protect the cell against cold stress and DNA damage. Extender containing 15% DMF with Ctrl, Bicine and egg yolk were the non-permeating cryoprotectants with higher post-thaw quality. The use of these compounds results in a reduction in vertebral fusions, compressions and severity of skeletal malformations in the offspring. Therefore, these extender compositions are beneficial for the quality of zebrafish offspring sired by cryopreserved sperm with 66 degrees C/min freezing rate. To the best of our knowledge, this is the first report on skeletal development of the offspring sired by cryopreserved sperm performed with different freezing media compositions in zebrafish.
dc.description.sponsorshipPortuguese Foundation for Science and Technology (FCT)Portuguese Foundation for Science and Technology [SFRH/BD/97466/2013]
dc.description.sponsorshipEuropean National funds (ASSEMBLE+) [730984]
dc.description.sponsorshipPortuguese national funds from FCT - Foundation for Science and Technology [UID/Multi/04326/2019]
dc.description.sponsorshipOperational programme CRESC Algarve 2020
dc.description.sponsorshipOperational programme COMPETE 2020 [EMBRC.PT ALG-01-0145-FEDER-022121]
dc.identifier.doi10.1016/j.cryobiol.2019.10.001
dc.identifier.issn0011-2240
dc.identifier.issn1090-2392
dc.identifier.urihttp://hdl.handle.net/10400.1/14156
dc.language.isoeng
dc.peerreviewedyes
dc.publisherAcademic Press inc Elsevier
dc.relationAssociation of European Marine Biological Laboratories Expanded
dc.subjectIn-vitro fertilization
dc.subjectSteroid glucuronides
dc.subjectOvarian fluid
dc.subjectAmino-acids
dc.subjectDna-damage
dc.subjectRerio
dc.subjectMotility
dc.subjectSpermatozoa
dc.subjectStorage
dc.subjectRepair
dc.titleCryoprotectants synergy improve zebrafish sperm cryopreservation and offspring skeletogenesis
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleAssociation of European Marine Biological Laboratories Expanded
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F97466%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/730984/EU
oaire.citation.endPage127
oaire.citation.startPage115
oaire.citation.titleCryobiology
oaire.citation.volume91
oaire.fundingStreamSFRH
oaire.fundingStreamH2020
person.familyNameDiogo
person.familyNameMartins
person.familyNameMarreiros
person.familyNameGavaia
person.familyNameCabrita
person.givenNamePatricia
person.givenNameGil
person.givenNameAna
person.givenNamePaulo
person.givenNameElsa
person.identifier.ciencia-idFD18-ED81-FFB5
person.identifier.ciencia-idD11F-8D8A-8EA1
person.identifier.ciencia-id9A12-9450-7051
person.identifier.ciencia-idB619-FC16-D007
person.identifier.orcid0000-0002-0804-5014
person.identifier.orcid0000-0002-1344-2954
person.identifier.orcid0000-0001-9410-4772
person.identifier.orcid0000-0002-9582-1957
person.identifier.orcid0000-0003-3864-8841
person.identifier.ridA-6470-2011
person.identifier.ridE-8523-2012
person.identifier.scopus-author-id36695981300
person.identifier.scopus-author-id57194785077
person.identifier.scopus-author-id6507104377
person.identifier.scopus-author-id26645697000
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameEuropean Commission
rcaap.rightsrestrictedAccess
rcaap.typearticle
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