Targeted Liver Ablation: Inducing Hepatocyte Death with Metronidazole in Transgenic Zebrafish Larvae
Published: April 30, 2023
Abstract
Source: Choi T.Y., et al. Hepatocyte-specific Ablation in Zebrafish to Study Biliary-driven Liver Regeneration. J. Vis. Exp. (2015)
The article describes targeted liver ablation, a method for inducing hepatocyte death with metronidazole in transgenic zebrafish larvae. The sample protocol describes the process for generating, manipulating and assessing targeted liver ablation in zebrafish.
Protocol
1. Preparation of Embryos/larvae Transfer 100 embryos into a 100 mm petri dish with 25 ml of egg water. Keep no more than 100 embryos per petri dish and raise them at 28 °C. To inhibit pigmentation, add phenylthiourea (PTU) into the petri dishes prior to 10 hr post-fertilization (hpf), and raise embryos/larvae until 80 hpf. The final concentration of PTU is 0.2 mM. CAUTION: PTU is toxic. Use in accordance with appropriate handling guidelines. Anesthetize t…
Materials
| 3-amino benzoic acid ethyl ester (Tricaine) powder | Sigma-Aldrich | A5040 | Make 0.4% (15 mM) tricaine stock : Bring 0.4 g tricaine to 100 mL with egg water. Adjust to pH 7. |
| Dimethyl sulfoxide (DMSO) | Sigma-Aldrich | D8418 | |
| Metronidazole | Sigma-Aldrich | M1547 | |
| N-Phenylthiourea (PTU) | Sigma-Aldrich | P7629 | |
| 100 x 15 mm petri dish | Denville Scientific Inc. | M5300 | |
| egg water | 0.3 g/L of sea salts ‘Instant Ocean’ and 0.5 mM CaSO4 in distilled water. | ||
| epifluorescence microscope | Leica Microsystems | M205FA |
Cite This Article
Targeted Liver Ablation: Inducing Hepatocyte Death with Metronidazole in Transgenic Zebrafish Larvae. J. Vis. Exp. (Pending Publication), e20181, doi: (2023).