IMAGE

Fig. 3

ID
ZDB-IMAGE-181026-9
Source
Figures for Gurevich et al., 2018
Image
Figure Caption

Fig. 3

Macrophages are necessary for wound angiogenesis and act to stabilise wound blood vessel sprouting tips

Representative confocal projection images of needle‐stick injured Tg(fli:GFP); Tg(mpeg:mCherry) or Tg(fli:GFP); Tg(mpeg:KalTA4); Tg(UAS:nfsB‐mCherry) zebrafish in combination with various treatments (as indicated), for ablating macrophages (red) during the peak stage of vessel (green) sprouting (4 DPI). Boxed area denotes wound site. In control (DMSO vehicle) and metronidazole‐treated fish lacking nitroreductase expressing macrophages, a normal inflammatory response and wound angiogenesis is observed. Upon ablation of macrophages using the nitroreductase–metronidazole system, wound angiogenesis is almost completely blocked. The same is true when macrophages are ablated using clodronate liposome injections.

  • Quantification of total blood vessel length in the presence/absence of macrophages at the wound site, measured from images represented in (A), using Angioanalyser. N = 16 independent fish per condition. Statistical significance, as determined by one‐way ANOVA, is P ≤ 0.0001. Subsequent Bonferroni multiple comparison test, determines level of significance, as indicated. Significance values: ***P ≤ 0.0001.

  • Images taken from timelapse movies of 4 DPF laser wounded Tg(fli:GFP); Tg(mpx:GFP); Tg(mpeg:mCherry) control or Tg(fli:GFP); Tg(mpx:GFP); Tg(mpeg:KalTA4); Tg(UAS:nfsB‐mCherry) macrophage ablation transgenic zebrafish, treated with metronidazole and imaged 30–930 min post‐injury (MPI). Boxed area denotes wound site. Arrowheads indicate position of damaged, sprouting vessel tips.

  • Quantification of proportion of vessel repaired within 930 MPI, measured from movies represented in (C). N = 8 independent fish per condition. Statistical significance is indicated, as determined by two‐tailed t‐test.

  • Graphical representation of typical cellular interaction events and their time course, measured from movies represented in (C). N = 8 independent fish per condition.

  • Gene expression of FAC‐sorted neutrophils extracted from Tg(mpx:GFP); Tg(mpeg:mCherry) needle‐stick wounded fish at 12 HPI, showing relative gene expression for sflt1 compared to unwounded fish at 4.5 DPF. N = 50 independent fish per condition per replicate, 3 replicates. Statistical significance is indicated, as determined by two‐tailed t‐test.

  • Gene expression of FAC‐sorted macrophages from Tg(mpx:GFP); Tg(mpeg:mCherry) transgenic zebrafish at 12 HPI post‐needle‐stick injury, showing relative gene expression for cyba1 and yrk compared to unwounded fish at 4.5 DPF. N = 50 independent fish per condition per replicate, 3 replicates. Statistical significance is indicated, as determined by two‐tailed t‐test.

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    This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ EMBO J.