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Bioluminescence Monitoring of Neuronal Activity in Freely Moving Zebrafish Larvae.


ABSTRACT: 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.

SUBMITTER: Knafo S 

PROVIDER: S-EPMC5681248 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature

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Bioluminescence Monitoring of Neuronal Activity in Freely Moving Zebrafish Larvae.

Knafo Steven S   Prendergast Andrew A   Thouvenin Olivier O   Figueiredo Sophie Nunes SN   Wyart Claire C  

Bio-protocol 20170901 18


The proof of concept for bioluminescence monitoring of neural activity in zebrafish with the genetically encoded calcium indicator <i>GFP-aequorin</i> has been previously described (Naumann <i>et al</i>., 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  ...[more]

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