ZFIN Image: Bruton et al., 2022, Fig. 7

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Figure Caption

Fig. 7

Figure 7. Vegfaa drives cardiomyocyte proliferation by endocardial notch signaling LSFM images of Tg(myl7:h2b-GFP) larvae at 24 hpi treated with PBS 0.1% BSA or zfVegfaa 0.1% BSA injection.

(B) Ventricular cardiomyocyte number in Tg(myl7:h2b-GFP) larvae at 24 and 48 hpi treated with PBS 0.1% BSA or zfVegfaa 0.1% BSA injection, n = 20. Unpaired t test.

(C) Images of injured ventricles from Tg(myl7:h2b-GFP) larvae, EdU stained and bathed in vehicle or AV951, imaged at 48 hpi. Non-myocardial EdU signal is excluded post-acquisitionally.

(D) Percentage of EdU+ cardiomyocyte nuclei from uninjured and injured ventricles from Tg(myl7:h2b-GFP) larvae, EdU stained and bathed in vehicle or AV951, n = 13–36. Unpaired t test.

(E) Images of injured Tg(myl7:nlsDsRed) larvae treated with vehicle or DAPT, acquired at 48 hpi by LSFM.

(F) Ventricular cardiomyocyte number in uninjured and injured Tg(myl7:h2b-GFP) larvae at 48 hpi treated with vehicle or DAPT, n = 24–40. Unpaired t test.

(G) Images of injured Tg(myl7:nlsDsRed) larvae treated with vehicle or AG1478, acquired at 48 hpi by LSFM.

(H) Ventricular cardiomyocyte number in uninjured and injured Tg(myl7:h2b-GFP) larvae at 48 hpi treated with vehicle or AG1478, n = 24. Unpaired t test.

(I) Treatment strategy for the injection of uninjured larvae with zfVegfaa and continuous bathing in AG1478 solution.

(J) Hypothesized signaling pathway active in uninjured and injured larval hearts driving cardiomyocyte proliferation.

(K) LSFM-acquired z plane showing notch expression colocalizing with endocardium in Tg(Tp1:venus-PEST;kdrl:hsa.HRAS-mCherry), abbreviated in the figure to Tg(Tp1:venus-PEST;kdrl:mCherry). AG1478 abbreviated to AG; white box, zoom panel.

(L) Proportion of larvae with notch+ endocardium at 6, 24, and 48 hpt following zfVegfaa injection and bathing in AG1478, n = 28. Fisher’s exact test.

(M) Treatment strategy for the lasering of larvae and continuous bathing in AG1478 solution.

(N) Representative z plane images of uninjured, injured, and injured AG-treated ventricles from Tg(tp1:venus-PEST) larvae at 48 hpi. BA, bulbous arteriosus; AVV, atrioventricular valve; white arrowheads, laterally inhibited cardiomyocytes; cyan arrowheads, notch+ endocardium; cyan box, zoom panel. Fisher’s exact test.

(O) Proportion of larvae with notch+ endocardium at 6, 24, and 48 hpt following laser injury and bathing in AG1478, n = 18.

(P) Cardiomyocyte number at 48 hpi following injection with recombinant Vegfaa and continuous bathing in DAPT or AG1478, n = 22–25. One-way ANOVA followed by Holms-Sidak’s multiple comparison post-hoc tests. All images are maximum intensity projections of 3D LSFM stacks unless otherwise stated. Scale bars, 50 μm. Data are represented as mean ± SEM, ^∗ p ≤ 0.05, ^∗∗ p ≤ 0.01, ^∗∗∗ p ≤ 0.001, and ^∗∗∗∗p ≤ 0.0001.

Acknowledgments

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Reprinted from Developmental Cell, 57(12), Bruton, F.A., Kaveh, A., Ross-Stewart, K.M., Matrone, G., Oremek, M.E.M., Solomonidis, E.G., Tucker, C.S., Mullins, J.J., Lucas, C.D., Brittan, M., Taylor, J.M., Rossi, A.G., Denvir, M.A., Macrophages trigger cardiomyocyte proliferation by increasing epicardial vegfaa expression during larval zebrafish heart regeneration, 1512-1528.e5, Copyright (2022) with permission from Elsevier. Full text @ Dev. Cell