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. 2008 Sep 12;374(1):143-6.
doi: 10.1016/j.bbrc.2008.06.109. Epub 2008 Jul 9.

Characterization of isolated ventricular myocytes from adult zebrafish (Danio rerio)

Fabien Brette  1 , Guillermo Luxan, Caroline Cros, Hayley Dixey, Christopher Wilson, Holly A Shiels
Affiliations

Characterization of isolated ventricular myocytes from adult zebrafish (Danio rerio)

Fabien Brette et al. Biochem Biophys Res Commun. .

Abstract

The zebrafish is widely used for human related disease studies. Surprisingly, there is no information about the electrical activity of single myocytes freshly isolated from adult zebrafish ventricle. In this study, we present an enzymatic method to isolate ventricular myocytes from zebrafish heart that yield a large number of calcium tolerant cells. Ventricular myocytes from zebrafish were imaged using light and confocal microscopy. Myocytes were mostly rod shaped and responded by vigorous contraction to field electrical stimulation. Whole cell configuration of the patch clamp technique was used to record electrophysiological characteristics of myocytes. Action potentials present a long duration and a plateau phase and action potential duration decreases when increasing stimulation frequency (as observed in larger mammals). Together these results indicate that zebrafish is a species ideally suited for investigation of ion channels related mutation screening of cardiac alteration important in human.

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Figures

Fig. 1
Fig. 1
Appearance of adult zebrafish ventricular myocytes under high magnification. (A) Phase contrast image of an isolated myocyte used for patch clamp experiments. (B) Confocal image of a single cardiac myocyte stained with di-8-ANNEPS. Scale bar is 5 μm. All cells were bathed in 2 mM Ca physiological saline solution.
Fig. 2
Fig. 2
Action potential characteristics of isolated ventricular myocytes from zebrafish. (A) Representative action potentials recorded at 0.1 Hz (black trace), 1 Hz (gray trace), and 2 Hz (light gray trace) in the same cell. (B) Mean (±SEM) action potential duration at 25%, 50%, and 90% repolarisation (APD25, APD50, and APD90) at 0.1 Hz (black bars), 1 Hz (gray bars), and 2 Hz (light gray bars). Data are from 14 myocytes. ∗ Indicates P < 0.05 versus 0.1 Hz and # indicates P < 0.05 versus 1 Hz.
Fig. 3
Fig. 3
Na and Ca currents in isolated ventricular myocytes from zebrafish. (A) Representative membrane current recorded at −40 mV (Na current) and 0 mV (Ca current). The inset shows the voltage protocol. (B) Mean (±SEM) Na current density (left) and Ca current density. Data are from 12 myocytes.
Fig. 4
Fig. 4
Ca current–voltage relationship in isolated ventricular myocytes from zebrafish. (A) Representative Ca currents recorded from −40 to +50 mV in 10 mV steps. The inset shows the voltage protocol. (B) Mean (±SEM) Ca current–voltage relationship. Data are from 10 myocytes.
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