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.
Date: 2017
Source: Bio-protocol   7: e2550 (Journal)
Registered Authors: Prendergast, Andrew, Wyart, Claire
Keywords: Bioluminescence, Coelenterazine, Escape response, GFP-aequorin-opt, Zebrafish
MeSH Terms: none
PubMed: 29130058 Full text @ Bio Protoc
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.