ZFIN ID: ZDB-FIG-190107-12
Lian et al., 2018 - Alas1 is essential for neutrophil maturation in zebrafish. Haematologica   103(11):1785-1795 Full text @ Haematologica
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Protruding-mouth to Day 4

Fig. 6

alas1 deficiency caused impaired host immunity against E. coli infection. (A) Arrowhead indicates the site of bacteria injection. The imaged region is boxed. (B-E) The in vivo behavior of neutrophils against E. coli was monitored by confocal microscopy. eGFP+ E. coli were subcutaneously injected over one somite into 3-day post fertilization (dpf) sibling (upper panels) and alas1smu350/smu350 mutant (lower panels) larva of the Tg(lyz:DsRed) background. Neutrophil behavior was analyzed through live imaging at 0.5 hours post injection (hpi) (B), 3 hpi (C), 5 hpi (D), and 24 hpi (E). All images are maximum-intensity projections from 25 steps × 2 μm. Scale bars: 50 μm. (F) Bacterial burden of embryos injected with E. coli. Significantly more bacterial cells were detected in alas1smu350/smu350 mutants (gray column) compared with siblings (black column) at 5 and 24 hpi. Mean±Standard Deviation (SD), n>10 in each group; Mann-Whitney U test: ****P<0.0001, **P<0.01, ns: not significant. (G) Quantification of recruited DsRed+ neutrophils at infection sites in live embryos injected with E. coli. Significantly more neutrophils were observed at the infection sites of alas1smu350/smu350 mutants (gray column) compared with siblings (black column) at 24 hpi. Mean±SD; n>19 in each group; Student t-test: ****P<0.0001. nd: not detectable, ns: not significant. (H) Relative il1b expression assessed by RT-qPCR. il1b expression was up-regulated in alas1smu350/smu350 mutants (gray column) compared with siblings (black column) at 3, 5, and 24 hpi. Mean±SD; n=4 in each group, performed in triplicate. Expression levels were adjusted for trauma [phosphate buffered saline (PBS) injection only]. Statistical significance was determined using Student t-test: **P<0.01, *P<0.05. ns: not significant. (I) Relative cxcl8a expression assessed by RT-qPCR. cxcl8a expression was up-regulated in alas1smu350/smu350 mutants (gray column) compared with siblings (black column) at 3 and 5 hpi. Mean±SD; n=4 in each group, performed in triplicate. Expression levels were adjusted for trauma (PBS injection only). Statistical significance was determined using Student t-test: **P<0.01. ns: not significant.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cxcl8a WT bacterial treatment by injection: Escherichia coli Protruding-mouth whole organism RTPCR
Day 4 whole organism RTPCR
alas1sm350/sm350 bacterial treatment by injection: Escherichia coli Protruding-mouth whole organism RTPCR
Day 4 whole organism RTPCR
DsRed2 nz50Tg control Protruding-mouth neutrophil IFL
Day 4 neutrophil IFL
nz50Tg ; alas1sm350/sm350 control Protruding-mouth neutrophil IFL
Day 4 neutrophil IFL
il1b WT bacterial treatment by injection: Escherichia coli Protruding-mouth whole organism RTPCR
Day 4 whole organism RTPCR
alas1sm350/sm350 bacterial treatment by injection: Escherichia coli Protruding-mouth whole organism RTPCR
Day 4 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
alas1sm350/sm350 bacterial treatment by injection: Escherichia coli Protruding-mouth whole organism il1b expression increased amount, abnormal
Protruding-mouth whole organism cxcl8a expression increased amount, abnormal
Day 4 whole organism il1b expression increased amount, abnormal
nz50Tg ; alas1sm350/sm350 bacterial treatment by injection: Escherichia coli Protruding-mouth - Day 4 defense response to bacterium process quality, abnormal
Day 4 defense response to bacterium process quality, abnormal
Day 4 neutrophil increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Haematologica for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Haematologica