In-vitro recording of adult zebrafish heart electrocardiogram — A platform for pharmacological testing

Tsai, Chia Ti; Wu, Cho Kai; Chiang, Fu-Tien; Tseng, Chuen Den; Lee, Jen Kuang; Yu, Chih Chieh; Wang, Yi Chih; Lai, Ling‐Ping; Lin, Jiunn Lee; Hwang, Juey Jen

doi:10.1016/j.cca.2011.07.002

Cited by 50 publications

(45 citation statements)

References 18 publications

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“…However, the diagnostic potential of optical mapping is complicated by the need for an excised heart and by the current need for contractile inhibitors, such as blebbistatin. Simultaneous voltage mapping and ECG recording results from the excised heart are highly correlated (32), and ECG recordings from intact animals provide a valuable benchmark. ECG recordings in the intact zebrafish require precise lead placement and are prone to motion artifacts caused by respiration (24); techniques such as paralysis of gill movement increase recording quality.…”

Section: Discussionmentioning

confidence: 99%

Construction and use of a zebrafish heart voltage and calcium optical mapping system, with integrated electrocardiogram and programmable electrical stimulation

Lin

Craig

Lamothe

et al. 2015

American Journal of Physiology-Regulatory, Integrative and Comp

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Zebrafish are increasingly being used as a model of vertebrate cardiology due to mammalian-like cardiac properties in many respects. The size and fecundity of zebrafish make them suitable for large-scale genetic and pharmacological screening. In larger mammalian hearts, optical mapping is often used to investigate the interplay between voltage and calcium dynamics and to investigate their respective roles in arrhythmogenesis. This report outlines the construction of an optical mapping system for use with zebrafish hearts, using the voltage-sensitive dye RH 237 and the calcium indicator dye Rhod-2 using two industrial-level CCD cameras. With the use of economical cameras and a common 532-nm diode laser for excitation, the rate dependence of voltage and calcium dynamics within the atrial and ventricular compartments can be simultaneously determined. At 140 beats/min, the atrial action potential duration was 36 ms and the transient duration was 53 ms. With the use of a programmable electrical stimulator, a shallow rate dependence of 3 and 4 ms per 100 beats/min was observed, respectively. In the ventricle the action potential duration was 109 ms and the transient duration was 124 ms, with a steeper rate dependence of 12 and 16 ms per 100 beats/min. Synchronous electrocardiograms and optical mapping recordings were recorded, in which the P-wave aligns with the atrial voltage peak and R-wave aligns with the ventricular peak. A simple optical pathway and imaging chamber are detailed along with schematics for the in-house construction of the electrocardiogram amplifier and electrical stimulator. Laboratory procedures necessary for zebrafish heart isolation, cannulation, and loading are also presented.

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Section: Discussionmentioning

confidence: 99%

Construction and use of a zebrafish heart voltage and calcium optical mapping system, with integrated electrocardiogram and programmable electrical stimulation

Lin

Craig

Lamothe

et al. 2015

American Journal of Physiology-Regulatory, Integrative and Comp

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“…A recent report in ZF revealed that both chromanol 293B (I Ks blocker) and E4031 (I Kr blocker) decrease HR, suggesting that I Kr and I Ks are also present in the ZF SAN; however, the effect of these drugs on the AV node and AV conduction was not noted (40). Given the complexity of the nodal currents in ZF, the exact mechanism by which cooling profoundly affects the heart rate is not known and requires further investigation.…”

Section: Acute Temperature Effects On Heart Rate and Rhythm (Av Delay)mentioning

confidence: 97%

“…Similarities in heart rate (HR), electrocardiogram and atrial and ventricular action potential (AP) morphology have been reported (32,33,40), suggesting parallels in ion channel expression and cell-cell coupling. Although the maximum upstroke velocity in the ZF cardiac AP is slower than in humans, the resting membrane potential is similar, the AP upstroke is dominated by an inward sodium current (I Na ), the plateau phase is managed by an L-type calcium current (I Ca,L ), and the rapid component of the delayed rectifier potassium current (I Kr ) governs repolarization of the membrane (33).…”

mentioning

confidence: 94%

Optical mapping of the electrical activity of isolated adult zebrafish hearts: acute effects of temperature

Lin

Ribeiro

Ding

et al. 2014

American Journal of Physiology-Regulatory, Integrative and Comp

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The zebrafish (Danio rerio) has emerged as an important model for developmental cardiovascular (CV) biology; however, little is known about the cardiac function of the adult zebrafish enabling it to be used as a model of teleost CV biology. Here, we describe electrophysiological parameters, such as heart rate (HR), action potential duration (APD), and atrioventricular (AV) delay, in the zebrafish heart over a range of physiological temperatures (18-28°C). Hearts were isolated and incubated in a potentiometric dye, RH-237, enabling electrical activity assessment in several distinct regions of the heart simultaneously. Integration of a rapid thermoelectric cooling system facilitated the investigation of acute changes in temperature on critical electrophysiological parameters in the zebrafish heart. While intrinsic HR varied considerably between fish, the ex vivo preparation exhibited impressively stable HRs and sinus rhythm for more than 5 h, with a mean HR of 158 ± 9 bpm (means ± SE; n = 20) at 28°C. Atrial and ventricular APDs at 50% repolarization (APD50) were 33 ± 1 ms and 98 ± 2 ms, respectively. Excitation originated in the atrium, and there was an AV delay of 61 ± 3 ms prior to activation of the ventricle at 28°C. APD and AV delay varied between hearts beating at unique HRs; however, APD and AV delay did not appear to be statistically dependent on intrinsic basal HR, likely due to the innate beat-to-beat variability within each heart. As hearts were cooled to 18°C (by 1°C increments), HR decreased by ~40%, and atrial and ventricular APD50 increased by a factor of ~3 and 2, respectively. The increase in APD with cooling was disproportionate at different levels of repolarization, indicating unique temperature sensitivities for ion currents at different phases of the action potential. The effect of temperature was more apparent at lower levels of repolarization and, as a whole, the atrial APD was the cardiac parameter most affected by acute temperature change. In conclusion, this study describes a preparation enabling the in-depth analysis of transmembrane potential dynamics in whole zebrafish hearts. Because the zebrafish offers some critical advantages over the murine model for cardiac electrophysiology, optical mapping studies utilizing zebrafish offer insightful information into the understanding and treatment of human cardiac arrhythmias, as well as serving as a model for other teleosts.

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“…The embryonic and adult zebrafish heart is proposed as an efficient platform for testing of drugs with potential electrophysiological effects on cardiomyocytes (Mittelstadt et al, 2008; Tsai et al, 2011) and for investigating human ion channel function in healthy and diseased state (Milan and Macrae, 2008). However, whether the zebrafish is truly an attractive alternative model for human cardiac electrophysiology depends on the presence and gating properties of the various ion channels in the zebrafish heart.…”

Section: Introductionmentioning

confidence: 99%

Zebrafish: a novel research tool for cardiac (patho)electrophysiology and ion channel disorders

Verkerk

Remme

2012

Front. Physio.

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The zebrafish is a cold-blooded tropical freshwater teleost with two-chamber heart morphology. A major advantage of the zebrafish for heart studies is that the embryo is transparent, allowing for easy assessment of heart development, heart rate analysis and phenotypic characterization. Moreover, rapid and effective gene-specific knockdown can be achieved using morpholino oligonucleotides. Lastly, zebrafish are small in size, are easy to maintain and house, grow fast, and have large offspring size, making them a cost-efficient research model. Zebrafish embryonic and adult heart rates as well as action potential (AP) shape and duration and electrocardiogram morphology closely resemble those of humans. However, whether the zebrafish is truly an attractive alternative model for human cardiac electrophysiology depends on the presence and gating properties of the various ion channels in the zebrafish heart, but studies into the latter are as yet limited. The rapid component of the delayed rectifier K+ current (IKr) remains the best characterized and validated ion current in zebrafish myocytes, and zebrafish may represent a valuable model to investigate human IKr channel-related disease, including long QT syndrome. Arguments against the use of zebrafish as model for human cardiac (patho)electrophysiology include its cold-bloodedness and two-chamber heart morphology, absence of t-tubuli, sarcoplamatic reticulum function, and a different profile of various depolarizing and repolarizing ion channels, including a limited Na+ current density. Based on the currently available literature, we propose that zebrafish may constitute a relevant research model for investigating ion channel disorders associated with abnormal repolarization, but may be less suitable for studying depolarization disorders or Ca2+-modulated arrhythmias.

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In-vitro recording of adult zebrafish heart electrocardiogram — A platform for pharmacological testing

Cited by 50 publications

References 18 publications

Construction and use of a zebrafish heart voltage and calcium optical mapping system, with integrated electrocardiogram and programmable electrical stimulation

Construction and use of a zebrafish heart voltage and calcium optical mapping system, with integrated electrocardiogram and programmable electrical stimulation

Optical mapping of the electrical activity of isolated adult zebrafish hearts: acute effects of temperature

Zebrafish: a novel research tool for cardiac (patho)electrophysiology and ion channel disorders

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