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. 2020 Oct 19;8:579943. doi: 10.3389/fcell.2020.579943

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Copyright © 2020 Kaveh, Bruton, Buckley, Oremek, Tucker, Mullins, Taylor, Rossi and Denvir.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Heart laser injury causes localized cardiomyocyte cell death. (A) Whole larva brightfield image of 3 dpf zebrafish with heart region indicated (black square), scale bar = 500 μm. (B) Brightfield images of a 3 dpf larval heart pre-laser injury (left) and 1-min post laser injury (right). The lesion is seen as a thickened and partially collapsed region at the ventricular apex (black arrowhead). (C) Ventricular ejection fraction before injury, and 2 hpi in injured and uninjured larvae. Error bars = SEM, n = 15–20 larvae, experimental n = 3. Unpaired t-test performed between groups where **** p < 0.0001. (D) Epifluorescence images of an uninjured and injured Tg(myl7:GFP) heart. Injury site is marked by a loss of myocardial GFP at the ventricular apex (white dashed line and arrowhead). (E) 3D LSFM image of a TUNEL stained injured Tg(myl7:h2b-GFP) heart at 2 hpi. Injury site is marked by a loss of nuclear myocardial GFP (white arrowhead) bordered by TUNEL positive cells (magenta). Image displayed as a maximum intensity projection (MIP). (F) LSFM single z-plane image of a Tg(myl7:mCherry;nfkb:GFP) ventricle at 2 and 24 hpi following heart injury. White arrowheads indicate loss of myocardial signal at 2 hpi and upregulation of nfkb:GFP in wound-bordering cardiomyocytes at 24 hpi. (G) 3D LSFM image of Tg(nfkb:GFP) ventricular expression at 24 hpi in uninjured and injured larvae (black arrowhead indicates ventricular apex injury site). Image displayed as a MIP (inverse color map). All scale bars = 50 μm unless stated otherwise. V, ventricle; A, atrium; ns, non-significant.