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First published online March 2, 2006
Journal of Experimental Biology 209, 1093-1100 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02117
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Development of the sympatho-vagal balance in the cardiovascular system in zebrafish (Danio rerio) characterized by power spectrum and classical signal analysis

Thorsten Schwerte*, Caroline Prem, Anita Mairösl and Bernd Pelster

Institute of Zoology and Limnology and Centre for Molecular Biosciences, University of Innsbruck, Innsbruck, Austria


Figure 1
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  		Fig. 1. The method of heart rate analysis using high speed video imaging. For analysis of the short-term heart rate variability (HRV) images (frames) of the beating heart were taken with 1000 or 200 frames s–1 and stored on the frame grabber's on-board memory. One sample image is shown in (A). Dotted lines indicate the perimeter of the atrium (a) and the ventricle (v). For each image a line was drawn beginning outside the heart tissue, cross sectioning the centre of ventricle and atrium and ending in the sinus venosus (solid line). Luminance profiles along these lines (see B), were stored for all frames. The intersection of the red lines marks the point on the line, where grey values through all frames are compiled to a luminance periodogram (C). The detected peaks (blue squares in C) were used for the calculation of inter-beat frequency (Hz) and plotted against frames in the tachogram (D). Red circles are the unfiltered raw data of every individual beat, and black rectangles represent the median filtered signal (median ranking=10). These data were taken for further analysis.

 

Figure 2
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  		Fig. 2. Absolute changes in heart rate after application of 1 µl (A) acetylcholine, atropine, (B) isoproterenol or propranolol (final concentration 10–4 mol l–1). Asterisks indicate statistically significant changes compared to control values (P<0.05). N=7 (per stage).

 

Figure 3
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  		Fig. 3. Typical power spectra of unfiltered luminance change frequencies in in situ recordings of zebrafish hearts in (A) 4 d.p.f. and (B) 15 d.p.f. zebrafish.

 

Figure 4
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  		Fig. 4. Typical data for heart rate variability in (A) 4 d.p.f. and (B) 15 d.p.f. zebrafish. The red circles are beat-to-beat frequencies of individual heart beat. The black circles are median values of the individual beats (median ranking=10).

 

Figure 5
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  		Fig. 5. Absolute changes in heart rate variability (HRV) frequency bandwidth, in control animals and in animals after incubation with (A) propranolol (ß-adrenergic antagonist, 10–5 mol l–1) and atropine (muscarinergic antagonist; 10–5 mol l–1) or (B) with each agonist separately. Asterisks indicate statistically significant differences between incubated and control animals (P<0.05). N=7 (per stage).

 




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