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Optimizing spinal cord injury in zebrafish larvae: effects of age on the injury response

View ORCID ProfileKirsten L. Underwood, View ORCID ProfileWhitney J. Walker, View ORCID ProfilePatrick I. Garrett, Shannon Linch, View ORCID ProfileThomas P. Rynes, View ORCID ProfileKaren Mruk
doi: https://doi.org/10.1101/2023.05.18.541337
Kirsten L. Underwood
1School of Pharmacy, University of Wyoming, Laramie, WY, USA
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Whitney J. Walker
1School of Pharmacy, University of Wyoming, Laramie, WY, USA
2Zoology and Physiology Graduate Program, University of Wyoming, Laramie, WY, USA
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Patrick I. Garrett
3Neurosciences Graduate Program, University of Wyoming, Laramie, WY, USA
4Department of Pharmacology and Toxicology, Brody School of Medicine, East Carolina University, Greenville, North Carolina, USA
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Shannon Linch
1School of Pharmacy, University of Wyoming, Laramie, WY, USA
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Thomas P. Rynes
1School of Pharmacy, University of Wyoming, Laramie, WY, USA
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Karen Mruk
1School of Pharmacy, University of Wyoming, Laramie, WY, USA
3Neurosciences Graduate Program, University of Wyoming, Laramie, WY, USA
4Department of Pharmacology and Toxicology, Brody School of Medicine, East Carolina University, Greenville, North Carolina, USA
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  • For correspondence: mrukk23@ecu.edu
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ABSTRACT

Zebrafish are an increasingly popular model to study spinal cord injury (SCI) regeneration. The transparency of larval zebrafish makes them ideal to study cellular processes in real time. Standardized approaches, including age of injury, are not readily available making comparisons of the results with other models challenging. In this study, we systematically examined the response to spinal cord transection of larval zebrafish at three different ages (3-7 days post fertilization or dpf) to determine whether the developmental complexity of the central nervous system affects the overall response to SCI. We then used imaging and behavioral analysis to evaluate whether differences existed based on the age of injury. All ages of larval zebrafish upregulated the required genes for glial bridge formation, ctgfa and gfap, at the site of injury, consistent with studies from adult zebrafish. Though all larval ages upregulated factors required to promote glial bridging, young larval zebrafish (3 dpf) were better able to regenerate axons independent of the glial bridge, unlike older zebrafish (7 dpf). Consistent with this data, locomotor experiments demonstrated that some swimming behavior occurs independent of glial bridge formation, further highlighting the need for standardization of this model and recovery assays. Overall, we found subtle cellular differences based on the age of transection in zebrafish, underlining the importance of considering age when designing experiments aimed at understanding regeneration.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted May 18, 2023.
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Optimizing spinal cord injury in zebrafish larvae: effects of age on the injury response
Kirsten L. Underwood, Whitney J. Walker, Patrick I. Garrett, Shannon Linch, Thomas P. Rynes, Karen Mruk
bioRxiv 2023.05.18.541337; doi: https://doi.org/10.1101/2023.05.18.541337
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Optimizing spinal cord injury in zebrafish larvae: effects of age on the injury response
Kirsten L. Underwood, Whitney J. Walker, Patrick I. Garrett, Shannon Linch, Thomas P. Rynes, Karen Mruk
bioRxiv 2023.05.18.541337; doi: https://doi.org/10.1101/2023.05.18.541337

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