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Generating Widespread and Scalable Retinal Lesions in Adult Zebrafish by Intraocular Injection of Ouabain

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Axon Regeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2636))

Abstract

Zebrafish regenerate functional retinal neurons after injury. Regeneration takes place following photic, chemical, mechanical, surgical, or cryogenic lesions, as well as after lesions that selectively target specific neuronal cell populations. An advantage of chemical retinal lesion for studying the process of regeneration is that the lesion is topographically widespread. This results in the loss of visual function as well as a regenerative response that engages nearly all stem cells (Müller glia). Such lesions can therefore be used to further our understanding of the process and mechanisms underlying re-establishment of neuronal wiring patterns, retinal function, and visually mediated behaviors. Widespread chemical lesions also permit the quantitative analysis of gene expression throughout the retina during the period of initial damage and over the duration of regeneration, as well as the study of growth and targeting of axons of regenerated retinal ganglion cells. The neurotoxic Na+/K+ ATPase inhibitor ouabain specifically offers a further advantage over other types of chemical lesions in that it is scalable; the extent of damage can be targeted to include only inner retinal neurons, or all retinal neurons, simply by adjusting the intraocular concentration of ouabain that is used. Here we describe the procedure through which these “selective” vs. “extensive” retinal lesions can be generated.

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Acknowledgments

Retinal regeneration studies in our laboratories using the approaches described have been supported by NIH R01 EY012146 (DLS), NIH R01 EY030467 (DMM), and NIH R21 EY026814 (DLS and DMM). We are also grateful for support in the form of pilot grants and other funding from Idaho INBRE (NIH P20 GM103408). We thank past and present members of the Stenkamp and Mitchell laboratories, particularly Ruth Frey for zebrafish husbandry and Lindsey Barrett for the images shown in Fig. 3a, b.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Idaho, Moscow, ID, USA

    Diana M. Mitchell & Deborah L. Stenkamp

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  1. Diana M. Mitchell
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  2. Deborah L. Stenkamp
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Correspondence to Diana M. Mitchell or Deborah L. Stenkamp .

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  1. Department of Biology, Appalachian State University, Boone, NC, USA

    Ava J. Udvadia

  2. Office of Technology Development, Medical College of Wisconsin, Milwaukee, WI, USA

    James B. Antczak

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Mitchell, D.M., Stenkamp, D.L. (2023). Generating Widespread and Scalable Retinal Lesions in Adult Zebrafish by Intraocular Injection of Ouabain. In: Udvadia, A.J., Antczak, J.B. (eds) Axon Regeneration. Methods in Molecular Biology, vol 2636. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3012-9_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3012-9_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3011-2

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