Research Output

Heart on a Plate: Histological and Functional Assessment of Isolated Adult Zebrafish Hearts Maintained in Culture

  The zebrafish is increasingly used for cardiovascular genetic and functional studies. We present a novel protocol to maintain and monitor whole isolated beating adult zebrafish hearts in culture for long-term experiments. Excised whole adult zebrafish hearts were transferred directly into culture dishes containing optimized L-15 Leibovitz growth medium and maintained for 5 days. Hearts were assessed daily using video-edge analysis of ventricle function using low power microscopy images. High-throughput histology techniques were used to assess changes in myocardial architecture and cell viability. Mean spontaneous Heart rate (HR, min−1) declined significantly between day 0 and day 1 in culture (96.7±19.5 to 45.2±8.2 min−1, mean±SD, p = 0.001), and thereafter declined more slowly to 27.6±7.2 min−1 on day 5. Ventricle wall motion amplitude (WMA) did not change until day 4 in culture (day 0, 46.7±13.0 µm vs day 4, 16.9±1.9 µm, p = 0.08). Contraction velocity (CV) declined between day 0 and day 3 (35.6±14.8 vs 15.2±5.3 µms−1, respectively, p = 0.012) while relaxation velocity (RV) declined quite rapidly (day 0, 72.5±11.9 vs day 1, 29.5±5.8 µms−1, p = 0.03). HR and WMA responded consistently to isoproterenol from day 0 to day 5 in culture while CV and RV showed less consistent responses to beta-agonist. Cellular architecture and cross-striation pattern of cardiomyocytes remained unchanged up to day 3 in culture and thereafter showed significant deterioration with loss of striation pattern, pyknotic nuclei and cell swelling. Apoptotic markers within the myocardium became increasingly frequent by day 3 in culture. Whole adult zebrafish hearts can be maintained in culture-medium for up to 3 days. However, after day-3 there is significant deterioration in ventricle function and heart rate accompanied by significant histological changes consistent with cell death and loss of cardiomyocyte cell integrity. Further studies are needed to assess whether this preparation can be optimised for longer term survival.

  • Type:

    Article

  • Date:

    13 May 2014

  • Publication Status:

    Published

  • Publisher

    Public Library of Science (PLoS)

  • DOI:

    10.1371/journal.pone.0096771

  • Library of Congress:

    QL Zoology

  • Dewey Decimal Classification:

    571 Physiology & related subjects

  • Funders:

    The British Heart Foundation Research Excellence Award (CoRE)

Citation

Pieperhoff, S., Wilson, K. S., Baily, J., de Mora, K., Maqsood, S., Vass, S., …Denvir, M. A. (2014). Heart on a Plate: Histological and Functional Assessment of Isolated Adult Zebrafish Hearts Maintained in Culture. PLoS ONE. 9(5), e96771. doi:10.1371/journal.pone.0096771. ISSN 1932-6203

Authors

Copyright

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Keywords

Zebrafish, hearts, culture experiments, cardiac ventricles, heart rate, histology, Isoproterenol, apoptosis, myocardium,

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