Home › Cell Biology › A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System
Cell Biology JoVE (Open Access) Citable · DOI

A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System

DOI: 10.3791/50439-v
What you'll learn
  • ✓ Set up and execute the GRIZLY luciferase reporter assay in zebrafish larvae
  • ✓ Treat transgenic larvae with test compounds and record luminescence signals
  • ✓ Analyze dose-response data to quantify glucocorticoid signaling modulation
Protocol

We describe the procedure and data analysis of a chemical screening system for glucocorticoid stress hormone signaling using zebrafish larvae: the Glucocorticoid Responsive In vivo Zebrafish Luciferase activitY (GRIZLY) assay. The assay sensitively and specifically detects effects on glucocorticoid signaling by compounds that require metabolization or affect endogenous glucocorticoid production.

Difficulty
advanced
Total time
~4–6 hours per experimental batch (compound treatment through luminescence recording and analysis)
Model organism
Zebrafish (Danio rerio) larvae, transgenic GRIZLY line
Biosafety
BSL-1

Steps

1
Prepare zebrafish larvae and GRIZLY system

Overview of the GRIZLY transgenic zebrafish line and glucocorticoid-responsive luciferase reporter construct. Establish baseline understanding of the assay platform and its sensitivity to glucocorticoid signaling.

▶ 02:22
2
Execute chemical compound treatment protocol

Apply test compounds to zebrafish larvae at defined developmental stages and concentrations. Document treatment conditions, exposure duration, and larval staging to ensure consistent glucocorticoid signaling activation.

▶ 04:40
3
Measure drug-induced luminescence response

Record luciferase luminescence output from treated larvae using a plate reader or imaging system. Capture temporal kinetics of reporter activation following compound exposure.

▶ 07:09
4
Analyze dose-response and quantify signaling

Process raw luminescence data, normalize to background controls, and generate dose-response curves. Calculate EC₅₀ values and statistical parameters to determine compound potency and specificity for glucocorticoid signaling modulation.

▶ 07:52
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