|ZFIN ID: ZDB-PUB-171114-5|
Bioluminescence Monitoring of Neuronal Activity in Freely Moving Zebrafish Larvae
Knafo, S., Prendergast, A., Thouvenin, O., Figueiredo, S.N., Wyart, C.
|Source:||Bio-protocol 7: e2550 (Journal)|
|Registered Authors:||Prendergast, Andrew, Wyart, Claire|
|Keywords:||Bioluminescence, Coelenterazine, Escape response, GFP-aequorin-opt, Zebrafish|
|PubMed:||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.
ABSTRACTThe 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.