ZFIN ID: ZDB-FIG-201003-187
Houseright et al., 2020 - Cell type specific gene expression profiling reveals a role for complement component C3 in neutrophil responses to tissue damage. Scientific Reports   10:15716 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Term:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Protruding-mouth to Days 7-13

Figure 3

Global depletion of c3a.1 decreases neutrophil recruitment to, and subsequent survival of, localized bacterial infection. (A) RT-qPCR validation of c3a, c3b, and c5a orthologue expression in pooled WT and c3a.1−/− (sa31241) whole zebrafish larvae, normalized to WT expression for each gene and to ef1α, with data expressed as mean + /- SEM. Data comprise 3 independent experiments, performed in triplicate, n = 50 larvae per condition per experiment. (B) Experimental setup. c3a.1+/+ (n = 71, 1 hpw; 65, 6 hpw) and c3a.1−/− (n = 71, 1 hpw; 68, 6 hpw) larvae were inoculated with 1000 CFU Pseudomonas aeruginosa in the left otic vesicle and subsequently stained with Sudan Black B. The otic vesicle region (box) was imaged. (C) Representative images and (D) quantification of neutrophil recruitment following otic vesicle infection, with data expressed as mean + /- SEM. Each dot represents one larva; colors represent results of 3 independent experiments. ***p < 0.001 (E) c3a.1+/+ (n = 24) and c3a.1−/− (n = 32) larvae were infected with 7500 CFU Pseudomonas aeruginosa in the left otic vesicle and survival was tracked over 5 days post-infection. (F) To test whether survival was neutrophil-dependent, c3a.1+/+ (n = 16) and c3a.1−/− (n = 14) larvae with neutrophils that are mcherry-labeled and carry a mutation in rac2 rendering them migration-defective (Tg(mpx:rac2D57N-mcherry)) were infected with Pseudomonas aeruginosa as in C and survival was tracked over 5 days post-infection. E and F each comprise 3 independent experiments.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
c3a.1 WT standard conditions Protruding-mouth whole organism RTPCR
c3a.1sa31241/sa31241 standard conditions Protruding-mouth whole organism RTPCR
c3a.6 WT standard conditions Protruding-mouth whole organism RTPCR
c3a.1sa31241/sa31241 standard conditions Protruding-mouth whole organism RTPCR
c3b.1 WT standard conditions Protruding-mouth whole organism RTPCR
c3a.1sa31241/sa31241 standard conditions Protruding-mouth whole organism RTPCR
c3b.2 WT standard conditions Protruding-mouth whole organism RTPCR
c3a.1sa31241/sa31241 standard conditions Protruding-mouth whole organism RTPCR
c5 WT standard conditions Protruding-mouth whole organism RTPCR
c3a.1sa31241/sa31241 standard conditions Protruding-mouth whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT bacterial treatment by injection: Pseudomonas aeruginosa PAK Days 7-13 otic vesicle neutrophil increased amount, abnormal
c3a.1sa31241/sa31241 standard conditions Protruding-mouth whole organism c3a.1 expression decreased amount, abnormal
Protruding-mouth whole organism c3b.1 expression decreased amount, abnormal
Protruding-mouth whole organism c5 expression increased amount, abnormal
c3a.1sa31241/sa31241 bacterial treatment by injection: Pseudomonas aeruginosa PAK Days 7-13 otic vesicle neutrophil increased amount, ameliorated
Days 7-13 whole organism viability, abnormal
c3a.1sa31241/sa31241; zf307Tg bacterial treatment by injection: Pseudomonas aeruginosa PAK Days 7-13 whole organism viability, abnormal
zf307Tg bacterial treatment by injection: Pseudomonas aeruginosa PAK Days 7-13 whole organism viability, abnormal
Acknowledgments:
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