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ZIRC
ZFIN ID: ZDB-FIG-140325-2
Shiau et al., 2013 - An Anti-inflammatory NOD-like Receptor Is Required for Microglia Development. Cell Reports   5(5):1342-1352 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Long-pec to Pec-fin
PHENOTYPE:
Fish:
Observed In:
Stage Range: Long-pec to Pec-fin

Fig. 2

Aberrant Migration of Primitive Macrophages in nlrc3-like/ Mutants

(A) Diagram shows the stereotypical migration route of macrophages from yolk sac to embryo proper (arrows).

(B) Fluorescent images show abnormal active coalescence of yolk sac macrophages (mpeg1:EGFP+ in green, arrows) that overlap with apoptotic marker TUNEL in red in mutants (bottom panel; n = 12/12), which does not occur in wild-type (top panel; n = 11/11). Region of imaging is indicated by yellow box in (A).

(C) Trajectories of individual yolk sac macrophages are shown using the MTrackJ cell-tracking tool, where the end point is indicated by a solid-triangle marker. See Movies S1 and S2 for the time-lapse series.

(D) Time-lapse imaging shows the head region (red box in A) where macrophages are expected to migrate into the brain, using transient transgenesis of the mpeg1:EGFP construct labeling macrophages in green and the stable transgene kdrl:mCherry-CAAX to visualize the vasculature in red. Imaging shows macrophages entering the brain in wild-type siblings (top) but not mutant (bottom). The numbers indicate the time in minutes. In the top panels, the arrowhead points to a macrophage that has taken up residence in the brain to become a microglial cell; the arrow points to another macrophage migrating into the brain. In the bottom panels, the arrow points to a mutant macrophage that does not enter the brain and remains associated with vasculature (as demarcated by dotted lines). See Movies S3 and S4.

(E) Whole-mount in situ hybridization at 2.5 dpf shows macrophage mfap4+ and microglial apoe+ cells in wild-type heterozygous embryos (arrows), but not in mutants (arrows). Aberrant macrophage clusters form near cranial vasculature in mutants (arrowheads). MB, midbrain; HB, hindbrain.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
apoeb nlrc3lst73/+ standard conditions Pec-fin brain microglial cell ISH
nlrc3lst73/st73 standard conditions Pec-fin Not Detected brain microglial cell ISH
EGFP nlrc3lst73/+; gl22Tg standard conditions Long-pec ball macrophage IHC
Long-pec ball macrophage IFL
nlrc3lst73/st73; gl22Tg standard conditions Long-pec ball macrophage IFL
Long-pec ball macrophage IHC
mfap4 nlrc3lst73/+ standard conditions Pec-fin brain macrophage ISH
nlrc3lst73/st73 standard conditions Pec-fin Not Detected brain macrophage ISH
Pec-fin cranial vasculature macrophage ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
nlrc3lst73/st73 standard conditions Pec-fin brain lacks all parts of type macrophage, abnormal
nlrc3lst73/st73 standard conditions Pec-fin brain lacks all parts of type microglial cell, abnormal
nlrc3lst73/st73 standard conditions Pec-fin microglia development decreased occurrence, abnormal
nlrc3lst73/st73; gl22Tg standard conditions Long-pec ball macrophage aggregated, abnormal
Long-pec ball macrophage mislocalised, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.