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Hypoxia-induced retinopathy model in adult zebrafish

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Abstract

Hypoxia-induced vascular responses, including angiogenesis, vascular remodeling and vascular leakage, significantly contribute to the onset, development and progression of retinopathy. However, until recently there were no appropriate animal disease models recapitulating adult retinopathy available. In this article, we describe protocols that create hypoxia-induced retinopathy in adult zebrafish. Adult fli1:EGFP zebrafish are placed in hypoxic water for 3–10 d and retinal neovascularization is analyzed using confocal microscopy. It usually takes 11 d to obtain conclusive results using the hypoxia-induced retinopathy model in adult zebrafish. This model provides a unique opportunity to study kinetically the development of retinopathy in adult animals using noninvasive protocols and to assess therapeutic efficacy of orally active antiangiogenic drugs.

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Figure 1: Hypoxia setup for adult zebrafish.
Figure 2: Critical steps of dissecting and analyzing the retinal vasculature in adult zebrafish.
Figure 3: Hypoxia-induced retinal angiogenesis.
Figure 4: Critical steps of dissecting and flatting the adult zebrafish retina.

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Acknowledgements

This study was supported through research grants from the Swedish Research Council, the Swedish Cancer Foundation, the Karolinska Institute Foundation, the Karolinska Institute Distinguished Professor Award, the European Union Integrated Project of Metoxia (Project no. 222741) and the European Research Council (ERC) advanced grant ANGIOFAT (Project no 250021). Z.C. was supported by the Swedish Heart and Lung Foundation.

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Author notes

  1. Ziquan Cao, Lasse D Jensen and Pegah Rouhi: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Medicine and Health, Linkoping University, Linkoping, Sweden

    Ziquan Cao, Toste Länne & Erik Wahlberg

  2. Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden

    Lasse D Jensen, Pegah Rouhi, Kayoko Hosaka & Yihai Cao

  3. Marine Biological Laboratory, Biological Institute, University of Copenhagen, Helsingor, Denmark

    John F Steffensen

Authors
  1. Ziquan Cao
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  2. Lasse D Jensen
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  3. Pegah Rouhi
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  4. Kayoko Hosaka
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  5. Toste Länne
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  6. John F Steffensen
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  7. Erik Wahlberg
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  8. Yihai Cao
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Contributions

Y.C. designed the study. L.D.J., P.R. and Z.C. performed the experiments. L.D.J., P.R., Z.C. and Y.C. analyzed the data. K.H., T.L., J.F.S. and E.W. participated in designing and discussing this study. L.D.J., P.R., Z.C. and Y.C. wrote the article.

Corresponding author

Correspondence to Yihai Cao.

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Cao, Z., Jensen, L., Rouhi, P. et al. Hypoxia-induced retinopathy model in adult zebrafish. Nat Protoc 5, 1903–1910 (2010). https://doi.org/10.1038/nprot.2010.149

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