PUBLICATION
Optimized Aequorin Reconstitution Protocol to Visualize Calcium Ion Transients in the Heart of Transgenic Zebrafish Embryos In Vivo
- Authors
- Vicente, M., Salgado-Almario, J., Martínez-Sielva, A., Llopis, J., Domingo, B.
- ID
- ZDB-PUB-220714-7
- Date
- 2022
- Source
- Methods in molecular biology (Clifton, N.J.) 2524: 271-280 (Chapter)
- Registered Authors
- Domingo Moreno, Beatriz, Llopis, Juan Francisco, Vicente Ruiz, Manuel
- Keywords
- Aequorin, Calcium, Coelenterazine, Heart, Zebrafish embryo
- MeSH Terms
-
- Aequorin*/genetics
- Aequorin*/metabolism
- Animals
- Animals, Genetically Modified
- Calcium/metabolism
- Ions/metabolism
- Luminescent Proteins/genetics
- Luminescent Proteins/metabolism
- Zebrafish*/metabolism
- PubMed
- 35821478 Full text @ Meth. Mol. Biol.
Citation
Vicente, M., Salgado-Almario, J., Martínez-Sielva, A., Llopis, J., Domingo, B. (2022) Optimized Aequorin Reconstitution Protocol to Visualize Calcium Ion Transients in the Heart of Transgenic Zebrafish Embryos In Vivo. Methods in molecular biology (Clifton, N.J.). 2524:271-280.
Abstract
We introduce how to image calcium ion levels in the heart of zebrafish embryos and larvae up to 5 days post-fertilization with the photoprotein green fluorescent protein (GFP)-aequorin (GA) in the transgenic line Tg(myl7:GA). Incubation of the embryos with CTZ to obtain the functional photoprotein yields few emission counts, suggesting that, when the heart is beating, the rate of aequorin consumption is faster than that of the reconstitution with CTZ. In this chapter, we present an improved aequorin reconstitution protocol. We further describe the experimental procedure as well as the bioluminescence data analysis and processing.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping