Figure 6

Figure 6

Neutrophil Cx43 Is Required for Autonomous and Cooperative Neutrophil Calcium Fluxes

(A) Time-lapse images showing calcium fluxes in neutrophils (arrow) contacting other neutrophils or necrotic cells in control untreated, CBX-treated, or cx43 MO-treated Tg(lyz:GCamp6F) larvae or in Tg(lyz:GCamp6F)xTg(lyz:cx43DN-T2A-mCherry) larvae. Time in minutes is indicated relative to the start of the cell-cell contact. Scale bar represents 15 μm.

(B) Cartoon illustrates contacts between neutrophils and necrotic PI⁺ cells or between fluxing and non-fluxing neutrophils resulting or not in calcium flux transmission.

(C and D) Percentage of neutrophil-neutrophil contacts (C) or neutrophil-PI⁺ cell contacts (D) resulting in transmission of calcium fluxes. Data are from Tg(lyz:GCamp6F)xTg(lyz:tRFP-5LO) (C and D) or Tg(lyz:GCamp6F) (D) larvae. Contacts in which none of the cells is initially fluxing are not included. n = 8 control, 4 CBX-treated larvae, and 7 Cx43 morphants from 8, 2, and 3 experiments, respectively (C). n = 5 control, 5 CBX-treated larvae, and 5 Cx43 morphants from 5, 2, and 2 experiments, respectively (D). One-way ANOVA, Tukey’s multiple comparisons test, is shown.

(E) Percentage of contacts resulting in transmission of calcium fluxes in Tg(lyz:GCamp6F)xTg(lyz:cx43DN-T2A-mCherry) zebrafish larvae, positive (cx43 DN) or negative for the Cx43DN-T2A-mCherry transgene (control siblings: Ctr sib). n = 7 cx43 DN and n = 5 control siblings from 3 experiments; Mann-Whitney test.

(F) Time point of arrival of first neutrophil at the wound in neutrophils positive (cx43 DN) or negative for the Cx43DN-T2A-mCherry transgene (control siblings: Ctr sib). n = 7 cx43 DN and n = 5 control siblings from 3 experiments; Mann Whitney test.

Error bars represent SEM. ^∗∗p < 0.002, ^∗∗∗p < 0.0002. See also Video S6.