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Evolving cardiac conduction phenotypes in developing zebrafish larvae: implications to drug sensitivity.

ABSTRACT: Cardiac arrhythmias include problems with impulse formation and/or conduction abnormalities. Zebrafish (Danio rerio) is an emerging model system for studying the cardiac conduction system. However, real-time recording of the electrocardiogram remains a challenge. In the present study, we assessed the feasibility of recording electrical cardiogram (ECG) signals from the zebrafish larvae using the micropipette electrodes, and demonstrated the dynamic changes in ECG signals and their sensitivity to Amiodarone during the developmental stages. We observed that ECG signals revealed P waves and QRS complexes at 7 days postfertilization (dpf). T waves started to develop at 14 dpf. Distinct P waves, QRS complexes, and T waves were similar to those of adult zebrafish at 35 dpf, accompanied by a statistically significant decrease in QRS intervals (from 256 ± 16 ms at 7 dpf to 54 ± 6 ms, p < 0.01, n = 5). In response to Amiodarone, ECG signals showed QRS prolongation from 7 to 35 dpf (p < 0.05, n = 5). Hence, micropipette electrodes can be applied to detect evolving ECG signals from the developing zebrafish larvae, thus providing a noninvasive and nonparalyzing approach to investigate cardiac conduction phenotypes in response to genetic, epigenetic, or pharmacologic perturbation.

SUBMITTER: Yu F

PROVIDER: S-EPMC3005146 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature

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Evolving cardiac conduction phenotypes in developing zebrafish larvae: implications to drug sensitivity.

Yu Fei F Huang Jie J Adlerz Katrina K Jadvar Hossein H Hamdan Mohamed H MH Chi Neil N Chen Jau-Nian JN Hsiai Tzung K TK

Zebrafish 20101020 4

Cardiac arrhythmias include problems with impulse formation and/or conduction abnormalities. Zebrafish (Danio rerio) is an emerging model system for studying the cardiac conduction system. However, real-time recording of the electrocardiogram remains a challenge. In the present study, we assessed the feasibility of recording electrical cardiogram (ECG) signals from the zebrafish larvae using the micropipette electrodes, and demonstrated the dynamic changes in ECG signals and their sensitivity to ...[more]

PMID: 20958244

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Project description:RationaleThe cardiac conduction system (CCS) consists of distinct components including the sinoatrial node, atrioventricular node, His bundle, bundle branches, and Purkinje fibers. Despite an essential role for the CCS in heart development and function, the CCS has remained challenging to interrogate because of inherent obstacles including small cell numbers, large cell-type heterogeneity, complex anatomy, and difficulty in isolation. Single-cell RNA-sequencing allows for genome-wide analysis of gene expression at single-cell resolution.ObjectiveAssess the transcriptional landscape of the entire CCS at single-cell resolution by single-cell RNA-sequencing within the developing mouse heart.Methods and resultsWild-type, embryonic day 16.5 mouse hearts (n=6 per zone) were harvested and 3 zones of microdissection were isolated, including: Zone I-sinoatrial node region; Zone II-atrioventricular node/His region; and Zone III-bundle branch/Purkinje fiber region. Tissue was digested into single-cell suspensions, cells isolated, mRNA reverse transcribed, and barcoded before high-throughput sequencing and bioinformatics analyses. Single-cell RNA-sequencing was performed on over 22 000 cells, and all major cell types of the murine heart were successfully captured including bona fide clusters of cells consistent with each major component of the CCS. Unsupervised weighted gene coexpression network analysis led to the discovery of a host of novel CCS genes, a subset of which were validated using fluorescent in situ hybridization as well as whole-mount immunolabeling with volume imaging (iDISCO+) in 3 dimensions on intact mouse hearts. Further, subcluster analysis unveiled isolation of distinct CCS cell subtypes, including the clinically relevant but poorly characterized transitional cells that bridge the CCS and surrounding myocardium.ConclusionsOur study represents the first comprehensive assessment of the transcriptional profiles from the entire CCS at single-cell resolution and provides a characterization in the context of development and disease.

Dataset Information

Evolving cardiac conduction phenotypes in developing zebrafish larvae: implications to drug sensitivity.

Publications

Evolving cardiac conduction phenotypes in developing zebrafish larvae: implications to drug sensitivity.

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