Figure 2.

cxcl8 morphants display a reduced zebrafish neutrophil recruitment in acute inflammation. Tail fins of Tg(mpx:gfp)i114 previously microinjected with standard control morpholino (MO StdC), MO cxcl8-l1 and MO cxcl8-l2 were transected at 3 dpf. (A) Representative maximum intensity projections of 6 hours confocal time-lapse microscopy of 3 dpf Tg(mpx:gfp)i114 control and morphant larvae, acquired at 3 min intervals. This sequence is shown in Supplemental Movie 1. Scale bar=100 μm (B) Counts of fluorescent neutrophils at the wound were made at 1, 2, 4 and 6 hpw. Data are presented as means ± SEM (n=30 performed as 3 independent experiments). P values were calculated using two-way ANOVA and Bonferroni multiple comparison test. At 1 hpw no significant differences in neutrophil number were observed for both morphants in comparison to control larvae (P>0.05). For MO cxcl8-l1 significant decreases were observed at 2 hpw (P<0.01) and from 4 to 6 hpw (P<0.001). As for MO cxcl8-l2, we have observed a significant decrease from 2 to 6 hpw (P<0.001) (C) DIC and GFP wide field fluorescence microscope micrographs from 3 dpf control and morphant larvae. (D) Total neutrophil counts in whole-larvae for each condition. Each bar represents means ± SEM (n=20 performed as 2 independent experiments). P values were calculated using one-way ANOVA and Bonferroni multiple comparison test (P>0.05, no significant differences were observed).