ZFIN ID: ZDB-FIG-171020-8
Vincent et al., 2017 - Neutrophil derived LTB4 induces macrophage aggregation in response to encapsulated Streptococcus iniae infection. PLoS One   12:e0179574 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Condition:
Knockdown Reagent:
Anatomical Term:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Condition:
Knockdown Reagents:
Observed In:
Stage Range: Protruding-mouth to Days 7-13

Fig. 5

Neutrophil-specific expression of Lta4h rescues macrophage aggregation and correlates with host survival in Lta4h-deficient larvae.

(A) Schematic of the Tol2-lyz:lta4h-2a-mCherry construct that was injected into AB-WT embryos to generate the Tg(lyz:lta4h-2a-mCherry) transgenic line. (B) The transgenic Lta4h sequence is non-targetable by morpholino knockdown. Tg(lyz:lta4h-2a-mCherry) (lyz:lta4h) or Tg(mpx:mCherry) (WT) was crossed to Tg(mpeg1:dendra2) and injected with either the Ctrl MO or Lta4h MO and monitored for macrophage aggregation at 24 hpi. Representative 20X images of WT or lyz:lta4h larvae at 24 hpi following inoculation with 50 CFU WT S. iniae, in both control and Lta4h morphant larvae. Scale bar is 80 μm. (C) Quantification of the average total percent of larvae forming macrophage aggregates from (B). (D) Survival of WT or lyz:lta4h larvae in either control or Lta4h morphant larvae infected with 50 CFU S. iniae. Compared with WT Ctrl MO larvae (black solid line), WT Lta4h MO larvae (black dotted line) have worse survival (p = 0.00437). Compared with lyz:lta4h Ctrl MO larvae (magenta solid line), lyz:lta4h Lta4h MO larvae (magenta dotted line) do not have significantly worse survival. However, lyz:lta4h Lta4h MO larvae (magenta dotted line) have significantly better survival than WT Lta4h MO larvae (black dotted line, p = 0.0142). (E) Increased survival in lyz:lta4h is macrophage dependent. Survival of WT or lyz:lta4h larvae in either control or Irf8 morphant larvae (lacking macrophages) infected with 50 CFU S. iniae. Compared with lyz:lta4h Ctrl MO larvae (green solid line), lyz:lta4h Irf8 MO larvae (green dotted line) have significantly worse survival (p < 0.0001). Compared with WT Irf8 MO larvae (black dotted line), lyz:lta4h Irf8 MO larvae do not have a significant difference in survival. Data are statistically pooled from at least 3 independent experiments, each with 24 larvae per condition.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
Dendra2 uwm12Tg ; uwm36Tg bacterial treatment by injection: Streptococcus iniae Protruding-mouth to Day 4 macrophage IFL
uwm12Tg ; uwm36Tg + MO2-lta4h bacterial treatment by injection: Streptococcus iniae Protruding-mouth to Day 4 macrophage IFL
uwm12Tg ; uwm5Tg bacterial treatment by injection: Streptococcus iniae Protruding-mouth to Day 4 macrophage IFL
uwm12Tg ; uwm5Tg + MO2-lta4h bacterial treatment by injection: Streptococcus iniae Protruding-mouth to Day 4 macrophage IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Days 7-13 whole organism decreased life span, abnormal
AB + MO1-irf8 bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Days 7-13 whole organism decreased life span, exacerbated
AB + MO2-lta4h bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Days 7-13 whole organism decreased life span, exacerbated
uwm12Tg ; uwm36Tg bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Day 4 macrophage aggregated, abnormal
uwm12Tg ; uwm36Tg + MO2-lta4h bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Day 4 macrophage aggregated, abnormal
uwm12Tg ; uwm5Tg bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Day 4 macrophage aggregated, abnormal
uwm12Tg ; uwm5Tg + MO2-lta4h bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Day 4 macrophage aggregated, ameliorated
uwm36Tg bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Days 7-13 whole organism decreased life span, exacerbated
uwm36Tg + MO1-irf8 bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Days 7-13 whole organism decreased life span, exacerbated
uwm36Tg + MO2-lta4h bacterial treatment by injection: Streptococcus iniae Protruding-mouth - Days 7-13 whole organism decreased life span, exacerbated
Acknowledgments:
ZFIN wishes to thank the journal PLoS One for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS One