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Revisiting in vivo staining with alizarin red S - a valuable approach to analyse zebrafish skeletal mineralization during development and regeneration

2016-01-19Journal article Open Access
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dc.contributor.authorBensimon-Brito, A.
dc.contributor.authorCardeira Da Silva, João
dc.contributor.authorDionísio, Gisela
dc.contributor.authorHuysseune, Ann
dc.contributor.authorCancela, Leonor
dc.contributor.authorWitten, Paul
dc.date.accessioned2016-01-26T12:58:04Z
dc.date.available2016-01-26T12:58:04Z
dc.date.issued2016-01-19
dc.date.updated2016-01-19T17:03:59Z
dc.description.abstractBackground The correct evaluation of mineralization is fundamental for the study of skeletal development, maintenance, and regeneration. Current methods to visualize mineralized tissue in zebrafish rely on: 1) fixed specimens; 2) radiographic and μCT techniques, that are ultimately limited in resolution; or 3) vital stains with fluorochromes that are indistinguishable from the signal of green fluorescent protein (GFP)-labelled cells. Alizarin compounds, either in the form of alizarin red S (ARS) or alizarin complexone (ALC), have long been used to stain the mineralized skeleton in fixed specimens from all vertebrate groups. Recent works have used ARS vital staining in zebrafish and medaka, yet not based on consistent protocols. There is a fundamental concern on whether ARS vital staining, achieved by adding ARS to the water, can affect bone formation in juvenile and adult zebrafish, as ARS has been shown to inhibit skeletal growth and mineralization in mammals. Results Here we present a protocol for vital staining of mineralized structures in zebrafish with a low ARS concentration that does not affect bone mineralization, even after repetitive ARS staining events, as confirmed by careful imaging under fluorescent light. Early and late stages of bone development are equally unaffected by this vital staining protocol. From all tested concentrations, 0.01 % ARS yielded correct detection of bone calcium deposits without inducing additional stress to fish. Conclusions The proposed ARS vital staining protocol can be combined with GFP fluorescence associated with skeletal tissues and thus represents a powerful tool for in vivo monitoring of mineralized structures. We provide examples from wild type and transgenic GFP-expressing zebrafish, for endoskeletal development and dermal fin ray regeneration.pt_PT
dc.identifier.citationBMC Developmental Biology. 2016 Jan 19;16(1):2pt_PT
dc.identifier.doihttp://dx.doi.org/10.1186/s12861-016-0102-4pt_PT
dc.identifier.otherAUT: LCA00739
dc.identifier.urihttp://hdl.handle.net/10400.1/7498
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherBioMed Centralpt_PT
dc.rights.holderBensimon-Brito et al.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.titleRevisiting in vivo staining with alizarin red S - a valuable approach to analyse zebrafish skeletal mineralization during development and regenerationpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FMulti%2F04326%2F2013/PT
oaire.citation.issue1pt_PT
oaire.citation.startPage2pt_PT
oaire.citation.titleBMC Developmental Biologypt_PT
oaire.citation.volume16pt_PT
oaire.fundingStream5876
person.familyNameBensimon-Brito
person.familyNameCardeira da Silva
person.familyNameDionísio
person.familyNameCancela
person.givenNameAnabela
person.givenNameJoão
person.givenNameGisela
person.givenNameM. Leonor
person.identifier1460592
person.identifier.ciencia-id8419-63A7-D0E5
person.identifier.ciencia-idDC15-86F7-3AAC
person.identifier.ciencia-id321D-EE64-25CD
person.identifier.orcid0000-0003-1663-2232
person.identifier.orcid0000-0003-3722-8810
person.identifier.orcid0000-0001-8891-9908
person.identifier.orcid0000-0003-3114-6662
person.identifier.ridN-7291-2014
person.identifier.scopus-author-id36154529200
person.identifier.scopus-author-id55224020100
person.identifier.scopus-author-id15833954700
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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