Bioluminescence Monitoring of Neuronal Activity in Freely Moving Zebrafish Larvae
- Knafo, S., Prendergast, A., Thouvenin, O., Figueiredo, S.N., Wyart, C.
- Bio-protocol 7: e2550 (Journal)
- Registered Authors
- Prendergast, Andrew, Wyart, Claire
- Bioluminescence, Coelenterazine, Escape response, GFP-aequorin-opt, Zebrafish
- MeSH Terms
- 29130058 Full text @ Bio Protoc
Knafo, S., Prendergast, A., Thouvenin, O., Figueiredo, S.N., Wyart, C. (2017) Bioluminescence Monitoring of Neuronal Activity in Freely Moving Zebrafish Larvae. Bio-protocol. 7:e2550.
The proof of concept for bioluminescence monitoring of neural activity in zebrafish with the genetically encoded calcium indicator GFP-aequorin has been previously described (Naumann et al., 2010) but challenges remain. First, bioluminescence signals originating from a single muscle fiber can constitute a major pitfall. Second, bioluminescence signals emanating from neurons only are very small. To improve signals while verifying specificity, we provide an optimized 4 steps protocol achieving: 1) selective expression of a zebrafish codon-optimized GFP-aequorin, 2) efficient soaking of larvae in GFP-aequorin substrate coelenterazine, 3) bioluminescence monitoring of neural activity from motor neurons in free-tailed moving animals performing acoustic escapes and 4) verification of the absence of muscle expression using immunohistochemistry.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes