ZFIN ID: ZDB-FIG-191224-3
Villani et al., 2019 - Clearance by Microglia Depends on Packaging of Phagosomes into a Unique Cellular Compartment. Developmental Cell   49(1):77-88.e7 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Long-pec to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 4 Tracking of Phagosomes Reveals that These Shrink and Converge into One Cellular Compartment

(A) V-plot showing size distribution of phagosomes in wild-type (n = 27, n = 141) and blbNY007 (n = 27, n = 47). p value < 0.0001.

(B) Quantification of relative vesicle diameter over time in wild-type (n = 13) and blbNY007 (n = 16). t0 corresponds to first fusion even. Data are normalized against the initial size.

(C and D) Representative time-lapse of phagosomal tracking in wild-type (C) (see also Video S3, upper panel) and blbNY007 (D) (see also Video S3, lower panel). Microglia are labeled with Tg(mpeg:GFP-caax). Scale bar, 5 μm.

(E) Quantification of the size of newly formed phagosomes in wild-type (n = 13) and in blbNY007 (n = 16). Whiskers 5th/95th percentile, p value: not significant.

(F) SPIM time-lapse of 2.5 dpf blbNY007 microglia labeled with Tg(mpeg:GFP-caax). The red dot marks the growing vesicle. Red, yellow, and blue tracks show the trajectories of three incoming phagosomes. Time indicated in minutes. Scale bar, 10 μm. See also Video S5.

(G) High time resolution SPIM time-lapse of a representative fusion event in a 3 dpf blbNY007 microglia labeled with Tg(mpeg:GFP-caax). Scale bar, 5 μm.

(H and I) Representative examples of microglia labeled with Tg(mpeg:GFP-caax) in blbNY007 (H) and in blbNY007 injected with a morpholino against the a1-vATPase (I) to prevent vesicular fusion.

(J) Quantification of experiment in (H) and (I). Percentage of microglia with more than 1 vesicle per cell in blbNY007 (n = 15) and blbNY007 injected with a morpholino against the a1-vATPase (n = 21), whiskers: 5th/95th percentile, p value < 0.0001.

(K) SPIM time-lapse of wild-type microglia labeled in Tg(mpeg::GFP-caax). Tracking highlights fusion events. Scale bar, 5 μm. Time in minutes. See also Video S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP slc37a2t30301/t30301 ; zh901Tg standard conditions Long-pec microglial cell phagocytic vesicle membrane IFL
Long-pec microglial cell plasma membrane IFL
Protruding-mouth microglial cell phagocytic vesicle membrane IFL
Protruding-mouth microglial cell plasma membrane IFL
slc37a2t30301/t30301 ; zh901Tg + MO1-atp6v0a1a standard conditions Protruding-mouth microglial cell phagocytic vesicle membrane IFL
Protruding-mouth microglial cell plasma membrane IFL
zh901Tg standard conditions Long-pec microglial cell phagocytic vesicle membrane IFL
Long-pec microglial cell plasma membrane IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
slc37a2t30301/t30301 ; zh901Tg standard conditions Long-pec microglial cell normal size, normal
Long-pec microglial cell phagocytic vesicle normal diameter, normal
Protruding-mouth microglial cell phagocytic vesicle increased diameter, abnormal
Protruding-mouth microglial cell phagosome maturation decreased occurrence, abnormal
slc37a2t30301/t30301 ; zh901Tg + MO1-atp6v0a1a standard conditions Protruding-mouth microglial cell has extra parts of type microglial cell phagocytic vesicle, abnormal
Protruding-mouth microglial cell vesicle fusion decreased occurrence, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 49(1), Villani, A., Benjaminsen, J., Moritz, C., Henke, K., Hartmann, J., Norlin, N., Richter, K., Schieber, N.L., Franke, T., Schwab, Y., Peri, F., Clearance by Microglia Depends on Packaging of Phagosomes into a Unique Cellular Compartment, 77-88.e7, Copyright (2019) with permission from Elsevier. Full text @ Dev. Cell