PUBLICATION
Cardioluminescence in Transgenic Zebrafish Larvae: A Calcium Imaging Tool to Study Drug Effects and Pathological Modeling
- Authors
- Vicente, M., Salgado-Almario, J., Collins, M.M., Martínez-Sielva, A., Minoshima, M., Kikuchi, K., Domingo, B., Llopis, J.
- ID
- ZDB-PUB-211025-34
- Date
- 2021
- Source
- Biomedicines 9(10): (Journal)
- Registered Authors
- Collins, Michelle, Domingo Moreno, Beatriz, Llopis, Juan Francisco, Martínez Sielva, Antonio, Vicente Ruiz, Manuel
- Keywords
- aequorin, calcium, coelenterazine, heart, heart failure, imaging, zebrafish
- MeSH Terms
- none
- PubMed
- 34680411 Full text @ Biomedicines
Citation
Vicente, M., Salgado-Almario, J., Collins, M.M., Martínez-Sielva, A., Minoshima, M., Kikuchi, K., Domingo, B., Llopis, J. (2021) Cardioluminescence in Transgenic Zebrafish Larvae: A Calcium Imaging Tool to Study Drug Effects and Pathological Modeling. Biomedicines. 9(10):.
Abstract
Zebrafish embryos and larvae have emerged as an excellent model in cardiovascular research and are amenable to live imaging with genetically encoded biosensors to study cardiac cell behaviours, including calcium dynamics. To monitor calcium ion levels in three to five days post-fertilization larvae, we have used bioluminescence. We generated a transgenic line expressing GFP-aequorin in the heart, Tg(myl7:GA), and optimized a reconstitution protocol to boost aequorin luminescence. The analogue diacetylh-coelenterazine enhanced light output and signal-to-noise ratio. With this cardioluminescence model, we imaged the time-averaged calcium levels and beat-to-beat calcium oscillations continuously for hours. As a proof-of-concept of the transgenic line, changes in ventricular calcium levels were observed by Bay K8644, an L-type calcium channel activator and with the blocker nifedipine. The β-adrenergic blocker propranolol decreased calcium levels, heart rate, stroke volume, and cardiac output, suggesting that larvae have a basal adrenergic tone. Zebrafish larvae treated with terfenadine for 24 h have been proposed as a model of heart failure. Tg(myl7:GA) larvae treated with terfenadine showed bradycardia, 2:1 atrioventricular block, decreased time-averaged ventricular calcium levels but increased calcium transient amplitude, and reduced cardiac output. As alterations of calcium signalling are involved in the pathogenesis of heart failure and arrhythmia, the GFP-aequorin transgenic line provides a powerful platform for understanding calcium dynamics.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping