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dc.contributor.authorKhaliullina-Skultety, Helenaen
dc.contributor.authorZi Chao, Ngiamen
dc.contributor.authorHarris, Williamen
dc.date.accessioned2017-12-04T13:18:58Z
dc.date.available2017-12-04T13:18:58Z
dc.date.issued2017-06-26en
dc.identifier.issn1940-087X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/269886
dc.description.abstractHere, we introduce a novel system for hypoxia induction, which we developed to study the effects of hypoxia in aquatic organisms such as frog and zebrafish embryos. Our system comprises a chamber featuring a simple setup that is nevertheless robust to induce and maintain a specific oxygen concentration and temperature in any experimental solution of choice. The presented system is very cost-effective but highly functional, it allows induction and sustainment of hypoxia for direct experiments in vivo and for various time periods up to 48 h. To monitor and study the effects of hypoxia, we have employed two methods - measurement of levels of hypoxia-inducible factor 1alpha (HIF-1α) in whole embryos or specific tissues and determination of retinal stem cell proliferation by 5-ethynyl-2'-deoxyuridine (EdU) incorporation into the DNA. HIF-1α levels can serve as a general hypoxia marker in the whole embryo or tissue of choice, here embryonic retina. EdU incorporation into the proliferating cells of embryonic retina is a specific output of hypoxia induction. Thus, we have shown that hypoxic embryonic retinal progenitors decrease proliferation within 1 h of incubation under 5% oxygen of both frog and zebrafish embryos. Once mastered, our setup can be employed for use with small aquatic model organisms, for direct in vivo experiments, any given time period and under normal, hypoxic or hyperoxic oxygen concentration or under any other given gas mixture.
dc.description.sponsorshipThis work was supported by the Support from Wellcome Trust SIA Award 100329/Z/12/Z to W.A.H. and the DFG fellowship KH 376/1-1 awarded to H.K.
dc.languageengen
dc.publisherJoVE
dc.titleInduction of Hypoxia in Living Frog and Zebrafish Embryos.en
dc.typeArticle
prism.numbere55710en
prism.publicationDate2017en
prism.publicationNameJournal of Visualized Experimentsen
prism.volume124en
dc.identifier.doi10.17863/CAM.16774
rioxxterms.versionofrecord10.3791/55710en
rioxxterms.versionAM*
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2017-06-26en
dc.contributor.orcidHarris, William [0000-0002-9995-8096]
dc.identifier.eissn1940-087X
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idWellcome Trust (069413/Z/02/Z)
pubs.funder-project-idWellcome Trust (100329/Z/12/Z)


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