Targeted Liver Ablation: Inducing Hepatocyte Death with Metronidazole in Transgenic Zebrafish Larvae

- Dissolve metronidazole in egg water containing DMSO, a solvent, and add phenylthiourea, which prevents pigment formation in embryos. Transfer a few larvae of the appropriate age to treatment and control plates. Next, add the desired concentration of the metronidazole solution to the test plate and egg water containing DMSO to the control plate.

Let the larvae in both the plates grow at 28 degrees Celsius. The transgenic larvae produce nitroreductase only in hepatocytes, which converts the metronidazole into a cytotoxic drug, resulting in the death of only the targeted hepatocytes. After treatment, remove the metronidazole solution from the treatment plate and wash the embryos with egg water.

Add tricaine to anesthetize the larvae and examine them under an epifluorescence microscope for severe hepatocyte ablation-- the destruction of liver cells. You will observe a much smaller liver in treated individuals than in control group individuals. In the example protocol, we will use metronidazole for hepatocyte ablation in transgenic zebrafish larvae to study liver regeneration.

- Transfer up to 100 embryos into a 100 millimeter Petri dish containing 25 milliliters of egg water and plac