Short-term chilled storage of Zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation

2.50
Hdl Handle:
http://hdl.handle.net/10547/560929
Title:
Short-term chilled storage of Zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation
Authors:
Desai, Kunjan ( 0000-0002-4204-5931 ) ; Spikings, Emma ( 0000-0001-8387-2098 ) ; Zhang, Tiantian
Abstract:
As zebrafish embryos have never been cryopreserved, we developed a protocol to store zebrafish embryos (50% epiboly-5.3 hour post fertilization) for up to 18 h at 0°C. Initial experiments to optimize the cryoprotectant (CPA) solution demonstrated improved embryo hatching rate following chilling at 0°C for 18 h with 1 M MeOH+0.1 M sucrose (56 ± 5%) compared with other combinations of methanol (0.2-0.5 M) and sucrose (0.05-0.1 M). This combination of CPAs that protects against chilling injury was further tested to assess its impact on sox gene and protein expression. Significant decreases in sox3 gene expression were observed in hatched embryos that had been chilled for 18 h in 1 M MeOH+0.1 sucrose compared with non-chilled controls, however the expression of both sox2 and sox3 proteins was unaffected. Significant decreases in sox2 protein expression were, however, observed in embryos that had been chilled without CPAs and these embryos also had lower hatching rates than those chilled with the optimal CPA solution. We, therefore, conclude that the CPA combination of 1 M MeOH+0.1 M sucrose facilitates chilled storage of early stage (50% epiboly) zebrafish embryos for up to 18 h without compromising transcriptional response.
Affiliation:
Georgia Regents University; University of Bedfordshire; Bournemouth University
Citation:
Desai, K., Spikings, E. and Zhang, T.T. (2015) 'Short-term chilled storage of Zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation' Zebrafish 12(1), pp.111-120.
Publisher:
Mary Ann Liebert
Journal:
Zebrafish
Issue Date:
Feb-2015
URI:
http://hdl.handle.net/10547/560929
DOI:
10.1089/zeb.2013.0961
Additional Links:
http://online.liebertpub.com/doi/abs/10.1089/zeb.2013.0961
Type:
Article
Language:
en
ISSN:
1545-8547; 1557-8542
Appears in Collections:
Biomedical Science

Full metadata record

DC FieldValue Language
dc.contributor.authorDesai, Kunjanen
dc.contributor.authorSpikings, Emmaen
dc.contributor.authorZhang, Tiantianen
dc.date.accessioned2015-07-24T08:36:07Zen
dc.date.available2015-07-24T08:36:07Zen
dc.date.issued2015-02en
dc.identifier.citationDesai, K., Spikings, E. and Zhang, T.T. (2015) 'Short-term chilled storage of Zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservation' Zebrafish 12(1), pp.111-120.en
dc.identifier.issn1545-8547en
dc.identifier.issn1557-8542en
dc.identifier.doi10.1089/zeb.2013.0961en
dc.identifier.urihttp://hdl.handle.net/10547/560929en
dc.description.abstractAs zebrafish embryos have never been cryopreserved, we developed a protocol to store zebrafish embryos (50% epiboly-5.3 hour post fertilization) for up to 18 h at 0°C. Initial experiments to optimize the cryoprotectant (CPA) solution demonstrated improved embryo hatching rate following chilling at 0°C for 18 h with 1 M MeOH+0.1 M sucrose (56 ± 5%) compared with other combinations of methanol (0.2-0.5 M) and sucrose (0.05-0.1 M). This combination of CPAs that protects against chilling injury was further tested to assess its impact on sox gene and protein expression. Significant decreases in sox3 gene expression were observed in hatched embryos that had been chilled for 18 h in 1 M MeOH+0.1 sucrose compared with non-chilled controls, however the expression of both sox2 and sox3 proteins was unaffected. Significant decreases in sox2 protein expression were, however, observed in embryos that had been chilled without CPAs and these embryos also had lower hatching rates than those chilled with the optimal CPA solution. We, therefore, conclude that the CPA combination of 1 M MeOH+0.1 M sucrose facilitates chilled storage of early stage (50% epiboly) zebrafish embryos for up to 18 h without compromising transcriptional response.en
dc.language.isoenen
dc.publisherMary Ann Lieberten
dc.relation.urlhttp://online.liebertpub.com/doi/abs/10.1089/zeb.2013.0961en
dc.subjectzebrafishen
dc.subjectDanio rerioen
dc.subjectcryopreservationen
dc.titleShort-term chilled storage of Zebrafish (Danio rerio) embryos in cryoprotectant as an alternative to cryopreservationen
dc.typeArticleen
dc.contributor.departmentGeorgia Regents Universityen
dc.contributor.departmentUniversity of Bedfordshireen
dc.contributor.departmentBournemouth Universityen
dc.identifier.journalZebrafishen
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