Fig. 1

Gβγ signaling is required for wound-induced neutrophil migrationin vivo. 72-hpf Tg(mpx:GFP) embryos pretreated for 30 min with the small-molecule Gβγ inhibitor M158C (25 μM), its inactive compound M158D (25 μM), or 1% DMSO only (control), were wounded (a portion of the tail fin was excised). (A-C′) Snapshots from epifluorescence and bright-field movies of the tail region (right) immediately after wounding (0 h, A-C) and 2 h after wounding (2 h, A′-C′). (A′′-C′′) Representative tracks (multiple colors) delineate routes of migration of individual neutrophils. (D) Number of neutrophils that reach the wound site within 2 h (the number of embryos analyzed is shown). (E) Total and net speeds of migration, and chemotaxis index, of neutrophils in 1% DMSO- (68 neutrophils, 7 embryos), M158D- (45 neutrophils, 5 embryos), and M158C- (67 neutrophils, 9 embryos) treated embryos. (F) Rose diagrams showing the directions of neutrophil movement events relative to the direction of migration (along anteroposterior axis, red line with arrowhead). The direction of migration was analyzed every 2–3 min throughout the time-lapse period, and were grouped into 30° sectors, with the number of events in each sector indicated. The fraction of migration events oriented at an angle of ±30° with respect to the direction of migration is 60% of 750 events in 1% DMSO-treated neutrophils, 59% of 453 events in M158D-treated neutrophils, and 25% of 453 events in M158C-treated neutrophils (p < 0.001, by t-test, two-way ANOVA, for M158C vs DMSO; p < 0.001, by one-way ANOVA for all three groups). Yellow lines: areas of wounding; Red line with arrowhead: the direction of migration. #: p > 0.05 vs control, *: p < 0.001 vs control.