Fig. 3

Observing collagen deposition/remodelling following skin wounding. (A) Schematic to illustrate the location/depth and tissues/layers involved in larval skin wounding. (B) Single and multi-channel images of wounds made to the flanks of Tg(krt19:col1a2-GFP), Tg(krt19:tdTomatoCAAX) double positive fish indicate how the epidermis (red cells) has partly healed in 20?min and completely healed over the denuded surface by 2?h post injury (hpi), whilst collagen I (green) remains absent in this region; see also Movie 1. (C-I) Max projection confocal images of Tg(krt19:col1a2-GFP) transgenic fish unwounded and wounded prior to imaging at the specified timepoints post injury (dpi); inset in C shows 3D reconstruction indicating collagen layer at 1?dpi with the collagen wound margin wrinkled where tension in this matrix meshwork has been released (arrows). E shows high magnification view of the interface between orthogonal (O) collagen, wound (W) margin and newly deposited wispy collagen fibrils (arrowheads) of a 5?dpi fish. (J) Quantification of collagen alignment index (AI) over the period of repair. For each timepoint mean of n?=?7?12 wounds are plotted?±?SD. ns, not significant, **p?<?0.01, ***p?<?0.001, ****p?<?0.0001. (K) A 3D reconstructed, sub-epidermal view in the region of the repairing wound (red dotted line) indicates tethers extending both into muscle and between myosepta (asterisks). (L) Schematic to show tethers in addition to myosepta that remain at the healed wound site. For C-I Scale bars =?15?µm; inset in C and K, 1 unit=?24.68?µm.