PUBLICATION
A Drug-Inducible Transgenic Zebrafish Model for Myelinating Glial Cell Ablation
- Authors
- Karttunen, M.J., Lyons, D.A.
- ID
- ZDB-PUB-190302-6
- Date
- 2019
- Source
- Methods in molecular biology (Clifton, N.J.) 1936: 227-238 (Chapter)
- Registered Authors
- Karttunen, Marja, Lyons, David A.
- Keywords
- Demyelination, Drug-inducible, Live imaging, Transgenic line, Zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Demyelinating Diseases/chemically induced*
- Demyelinating Diseases/metabolism
- Disease Models, Animal
- Gene Expression Regulation, Developmental
- Metronidazole/pharmacology*
- Myelin Basic Protein/genetics
- Neuroglia
- Nitroreductases/genetics
- Nitroreductases/metabolism*
- Oligodendroglia/cytology*
- Oligodendroglia/drug effects
- Promoter Regions, Genetic
- Remyelination/drug effects
- Zebrafish/genetics
- Zebrafish/growth & development*
- PubMed
- 30820902 Full text @ Meth. Mol. Biol.
Citation
Karttunen, M.J., Lyons, D.A. (2019) A Drug-Inducible Transgenic Zebrafish Model for Myelinating Glial Cell Ablation. Methods in molecular biology (Clifton, N.J.). 1936:227-238.
Abstract
To study cellular and molecular mechanisms of demyelination and remyelination in vivo, we developed a transgenic zebrafish line, Tg(mbp:mCherry-NTR), in which expression of the bacterial enzyme nitroreductase (NTR) is driven under the myelin basic protein promoter (mbp) and thus is expressed in myelinating glia. When NTR-expressing larvae are treated with the prodrug metronidazole, the reaction between NTR and Mtz results in a toxic metabolite which selectively kills NTR-expressing cells. Using the Tg(mbp:mCherry-NTR) line, we can ablate two-thirds of oligodendrocytes following a 2-day MTZ treatment. Demyelination is evident seven days later, and remyelination is observed 16 days after Mtz treatment. The Tg(mbp:mCherry-NTR) model can be used to image cell behavior during, and to test how genetic manipulations or chemical compounds regulate, demyelination and remyelination. In this chapter, we describe the methods we used to characterize the oligodendrocyte loss, demyelination and remyelination in the Tg(mbp:mCherry-NTR) model.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping