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Updated: Aug 8, 2025
Analysis of Oxidative Stress in Zebrafish Embryos
Published on: July 7, 2014
Duox is the primary NADPH oxidase responsible for ROS production during adult caudal fin regeneration in zebrafish
Kunal Chopra1, Milda Folkmanaitė1, Liam Stockdale1
1Division of Cell Matrix Biology & Regenerative Medicine, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PT, UK.
Abstract:
Sustained elevated levels of reactive oxygen species (ROS) have been shown to be essential for regeneration in many organisms. This has been shown primarily via the use of pharmacological inhibitors targeting the family of NADPH oxidases (NOXes). To identify the specific NOXes involved in ROS production during adult caudal fin regeneration in zebrafish, we generated nox mutants for duox, nox5 and cyba (a key subunit of NOXes 1-4) and crossed these lines with a transgenic line ubiquitously expressing HyPer, which permits the measurement of ROS levels. Homozygous duox mutants had the greatest effect on ROS levels and rate of fin regeneration among the single mutants. However, duox:cyba double mutants showed a greater effect on fin regeneration than the single duox mutants, suggesting that Nox1-4 also play a role during regeneration. This work also serendipitously found that ROS levels in amputated adult zebrafish fins oscillate with a circadian rhythm.
Insights
Reactive oxygen species (ROS) are crucial for zebrafish fin regeneration. Specific NADPH oxidases (NOXes), particularly Duox, are key players, with ROS levels exhibiting a daily rhythm post-amputation.
Area of Science:
- Zebrafish fin regeneration
- Reactive oxygen species (ROS) biology
- Molecular mechanisms of regeneration
Background:
- Sustained elevated reactive oxygen species (ROS) levels are vital for regeneration across diverse organisms.
- Pharmacological inhibitors targeting NADPH oxidases (NOXes) have primarily demonstrated the role of ROS in regeneration.
- Understanding specific NOX family members involved in ROS production during fin regeneration is crucial.
Purpose of the Study:
- To identify specific NADPH oxidases (NOXes) responsible for ROS production during adult zebrafish caudal fin regeneration.
- To investigate the roles of Duox, Nox5, and Cyba in ROS-mediated fin regeneration.
- To analyze ROS level dynamics during the fin regeneration process.
Main Methods:
- Generation and utilization of specific nox mutants (duox, nox5, cyba) in zebrafish.
- Crossbreeding mutant lines with a transgenic HyPer reporter line for ROS level measurement.
- Assessment of ROS levels and fin regeneration rates in wild-type and mutant zebrafish.
Main Results:
- Homozygous duox mutants exhibited the most significant reduction in ROS levels and fin regeneration rate among single mutants.
- duox:cyba double mutants displayed impaired fin regeneration compared to single duox mutants, indicating a role for Nox1-4.
- ROS levels in amputated adult zebrafish fins were found to oscillate with a circadian rhythm.
Conclusions:
- Duox is a primary NOX enzyme involved in ROS production during zebrafish fin regeneration.
- Nox1-4 family members also contribute to fin regeneration, as evidenced by the enhanced effect in duox:cyba double mutants.
- The study revealed a novel circadian oscillation of ROS levels during zebrafish fin regeneration, opening new avenues for research.
