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. 2019 Aug 16;116(7):1357–1371. doi: 10.1093/cvr/cvz221

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© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society of Cardiology

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Individual immune cell lineages are predominant at different times post-injury. Representative images of whole hearts dissected from Tg(mpx:GFP) (A), Tg(lyz:dsRED2) (B), Tg(c-fms:GFP); Tg(mpeg1:mCherry) (D and E), Tg(lck:GFP) (G), and Tg(gata2a:GFP); Tg(lyz:dsRED2) (H) transgenic zebrafish at the time-points post-injury as indicated. The types of inflammatory cells labelled by the different transgenics are shown above the images. Quantification of the different inflammatory cells across a timeline from unwounded to 21 dpi (C, F, and I). In C the total L-plastin+ cells present at each timepoint is shown in blue. L-plastin+ data also shown in F and I to allow comparison to total inflammatory cell numbers (same data, grey in F and I). (A–C) Neutrophils [marked by Tg(mpx:GFP) (A) and Tg(lyz:dsRED2) (B)] peak at 6 hpi and resolve by 7 dpi (C). L-plastin labelling for all leucocytes (in magenta) is also shown in A. (D–F) Macrophages, marked by Tg(c-fms:GFP) and Tg(mpeg1:mCherry), are present in large numbers at all time-points and show significant infiltration compared to unwounded hearts at 7 dpi (F). The majority of macrophages express both transgenic reporters (inset in D). (G) Analysis of Tg(lck:GFP) fish reveals significant numbers of T-cells at 7 dpi.³³ Eosinophils, labelled with Tg(gata2a:GFP) (H), are rarely observed in unwounded hearts but are significantly recruited at 6 hpi (I) and at later time-points (7, 14, and 21 dpi) (I). Tg(gata2a:GFP); Tg(lyz:dsRED2) double transgenic fish demonstrate two distinct populations of cells present at 24 hpi (inset in H). Boxed regions demark the approximate position of the insets for each panel. Dashed lines in A, B, and E–H demark the injured region. In C, F, and I, colour coded n numbers are provided for each time-point and transgenic reporter. L-plastin n numbers are only shown in C for clarity. For statistical analyses Kruskal–Wallis/Dunn’s multiple comparisons tests were used to analyse all data against unwounded for each data set. All other values were not significant. Scale bars: A, B, D, E, G, H = 250 μm; insets = 50 μm.