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. 2022 Jun 20;57(12):1512–1528.e5. doi: 10.1016/j.devcel.2022.05.014

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© 2022 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Macrophages stimulate epicardial cell number expansion following cardiac injury

(A) Image of an uninjured 3 dpf ventricle from a Tg(myl7:mKateCAAX;myl7:h2b-GFP) larva showing vegfaa+ cells (green) overlying myocardium (magenta). Cyan box, zoom panel.

(B) Images of uninjured and injured ventricles from Tg(vegfaa:GFP) larvae acquired at 48 hpi. “Heat” LUT is applied to highlight the increased intensity of epicardial vegfaa:GFP in injured hearts.

(C) Total ventricular vegfaa:GFP fluorescence in uninjured and injured hearts over standard injury model time points, n = 28–30. ^∗ p ≤ 0.05 one-way ANOVA followed by Holms-Sidak’s multiple comparison post-hoc tests.

(D and E) Images of uninjured and injured EdU-stained Tg(vegfaa:GFP) hearts 48 hpi showing EdU+ epicardial cells (white arrowheads) and the proportion of EdU+ epicardial cells are quantified in (E), n = 13–16. Unpaired t test, ^∗∗ p ≤ 0.01.

(F) Image of a ventricle from a Tg(vegfaa:GFP;csfr1a:NfsB-mCherry;kdrl:hsa.HRAS-mCherry) (abbreviated to kdrl:mCherry) larva at 48 hpi showing macrophages in the epicardial-myocardial niche (white arrowheads). Cyan box, zoom panel.

(G) Images of uninjured and injured ventricles from Tg(vegfaa:GFP;csfr1a:NfsB-mCherry) larvae from metronidazole-nitroreductase macrophage ablation groups at 48 hpi. “Heat” LUT is applied to highlight the increase in the overall fluorescence in injured groups except NTR+met+. Total vegfaa:GFP fluorescence (H) and epicardial cell number (I) in uninjured and injured ventricles from Tg(vegfaa:GFP;csfr1a:NfsB-mCherry) larvae from metronidazole-nitroreductase macrophage ablation groups at 48 hpi. All images are maximum intensity projections of 3D LSFM stacks. Scale bars, 50 μm, n = 46. ^∗ p ≤ 0.05, ^∗∗ p ≤ 0.01, ^∗∗∗p ≤ 0.001. One-way ANOVA followed by Holms-Sidak’s multiple comparison post-hoc tests. Data are represented as mean ± SEM.