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Electroretinogram analysis of zebrafish retinal function across development

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Abstract

Purpose

The electroretinogram (ERG) is a powerful approach for investigating visual function in zebrafish ocular disease models. However, complexity, cost, and a literature gap present as significant barriers for the introduction of this technology to new zebrafish laboratories. Here, we introduce a simplified and effective method to obtain zebrafish ERGs.

Methods

In-house assembled recording electrodes and a custom 3D-printed platform were used to gather high-quality and consistent ERG data from zebrafish at 3 developmental timepoints—larval, juvenile, and adult. Fish were tested under both scotopic (dark-adapted) and photopic (light-adapted) conditions to differentiate between the rod and cone systems, respectively.

Results

Robust ERG waveforms across all developmental timepoints were obtained using the methodology presented here. We observed an overall increase in signal amplitude as development progressed, reflecting maturation of the zebrafish retina. Oscillatory potentials could also be isolated from the generated waveforms.

Conclusions

This simplified approach to the zebrafish ERG can generate waveforms comparable to the existing approaches and helps reduce barriers for zebrafish laboratories studying ocular development and disease.

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Availability of data and materials

The data used during this study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank Dr. Yves Sauvé and Dr. Orson Moritz for sharing their extensive technical skills and knowledge of ERG recordings. We would also like to thank Science Animal Support Services at the University of Alberta for their excellent care of the zebrafish aquatics facility.

Funding

This work was supported by an Alberta Vision Net Catalyst Grant (JCH), Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant (RGPIN-2018-05756, JCH), Alberta Graduate Excellence Scholarship (NJN), NSERC Canada Graduate Student—Master’s Grant (NJN), Queen Elizabeth II Graduate Scholarship (NJN), Walter H. Johns Graduate Fellowship (NJN), a Faculty of Medicine and Dentistry (University of Alberta)/Alberta Health Services Graduate Student Recruitment Studentship (NJN), a Department of Surgery (University of Alberta) Summer Studentship (CXLW), and an NSERC Undergraduate Student Research Award. The research was also funded by the generous support of the Stollery Children's Hospital Foundation through a Women and Children’s Health Research Institute Innovation Grant (WCHRIIG-2846, JCH) and a summer studentship (CXLW).

Author information

Authors and Affiliations

  1. Department of Medical Genetics, University of Alberta, Edmonton, Canada

    Nathan J. Nadolski & Jennifer C. Hocking

  2. Department of Biological Sciences, University of Alberta, Edmonton, AB, Canada

    Casey X. L. Wong

  3. Women & Children’s Health Research Institute, University of Alberta, Edmonton, AB, Canada

    Casey X. L. Wong & Jennifer C. Hocking

  4. Department of Cell Biology, University of Alberta, Edmonton, AB, Canada

    Jennifer C. Hocking

  5. Division of Anatomy, Department of Surgery, University of Alberta, 5-01 Medical Sciences Building, Edmonton, AB, T6G 2H7, Canada

    Jennifer C. Hocking

Authors
  1. Nathan J. Nadolski
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  2. Casey X. L. Wong
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  3. Jennifer C. Hocking
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Nathan Nadolski and Casey Wong. The first draft of the manuscript was written by Nathan Nadolski, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Jennifer C. Hocking.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Consent for publication

N.J. Nadolski, Yes; C.X.L. Wong, Yes; J.C. Hocking, Yes.

Ethical approval

Animal ethics protocols were approved by the University of Alberta Biosciences Animal Care Committee (AUP1476).

Informed consent

Informed consent was not applicable.

Statement of human rights

This article does not contain any studies with human participants performed by any of the authors.

Statement on the welfare of animals

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

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Electronic supplementary material

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Supplementary Figure 1

Scotopic b-wave amplitude comparisons across development for individual stimulus intensities. Scotopic b-wave amplitudes for each age cohort in response to a) -0.7 log cd•s/m2, b) 0.0 log cd•s/m2, and c) 1.0 log cd•s/m2 stimulus intensities are compared. Boxes extend from the 25-75% percentiles, internal lines represent the median, and whiskers complete the range. n=15 for 7 dpf, n=10 for 1 mpf, and n=10 for 4 mpf. *P<0.05; **P<0.01; ***P<0.001 (TIF 8337 kb)

Supplementary File 1

3D model of the ERG platform visualized in Figure 1b. This model can be downloaded as an .stl file, modified as necessary, and then 3D printed with polylactic acid (PLA) filament using the appropriate hardware (STL 57 kb)

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Nadolski, N.J., Wong, C.X.L. & Hocking, J.C. Electroretinogram analysis of zebrafish retinal function across development. Doc Ophthalmol 142, 99–109 (2021). https://doi.org/10.1007/s10633-020-09783-y

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  • DOI: https://doi.org/10.1007/s10633-020-09783-y

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