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Monitoring Nrf2/ARE Pathway Activity with a New Zebrafish Reporter System
Lorenzo Badenetti1,2, Rosa Manzoli3,4, Michela Rubin3
1Department of Women's and Children's Health, University of Padova, I-35128 Padova, Italy.
Abstract:
Among multiple cytoprotective mechanisms, eukaryotic cells exhibit a complex transcriptional program relying on the Nrf2 transcription factor, which is generally recruited upon biological stressors including oxidative-stress-based cellular insults. The relevance of this master regulator has remarkably emerged in recent years in several research fields such as cancer, inflammatory disorders and age-related neurological diseases. Here, we document the generation and characterization of a novel Nrf2/ARE pathway biosensor fish which exhibits a dynamic spatiotemporal expression profile during the early developmental stages. The transgenic line is responsive to known Nrf2 pathway modulators but also to Edaravone, which direct activity on the Nrf2 pathway has never been documented in a live transgenic fish model. We also show that the reporter is faithfully activated during fin regeneration, and its degree of expression is slightly affected in a glucocerebrosidase (Gba1) morphant zebrafish model. Therefore, this novel transgenic fish may represent a valuable tool to be exploited for the characterization of zebrafish models of human diseases, as well as for primary high-throughput drug screening.
Insights
Researchers developed a new Nrf2/ARE pathway biosensor fish. This tool tracks cellular stress responses and aids in drug screening for diseases like cancer and neurological disorders.
Area of Science:
- Molecular Biology
- Genetics
- Toxicology
Background:
- Eukaryotic cells utilize a transcriptional program involving the Nrf2 transcription factor for cytoprotection against stressors.
- The Nrf2 pathway is crucial in cancer, inflammatory disorders, and age-related neurological diseases.
Purpose of the Study:
- To generate and characterize a novel Nrf2/ARE pathway biosensor fish.
- To assess the biosensor's responsiveness to modulators and its utility in disease modeling and drug screening.
Main Methods:
- Generation of a transgenic Nrf2/ARE pathway biosensor fish.
- Analysis of spatiotemporal expression during development.
- Testing responsiveness to known Nrf2 modulators and Edaravone.
- Evaluation in fin regeneration and a glucocerebrosidase (Gba1) morphant model.
Main Results:
- The biosensor fish displays dynamic spatiotemporal expression during early development.
- The reporter is responsive to Nrf2 pathway modulators and Edaravone.
- Reporter activation was observed during fin regeneration.
- Expression was slightly affected in Gba1 morphant zebrafish.
Conclusions:
- The novel Nrf2/ARE biosensor fish is a valuable tool for studying Nrf2 pathway dynamics.
- This model can aid in characterizing zebrafish disease models and high-throughput drug screening.
