ZFIN ID: ZDB-FIG-191224-1
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:
Genes:
Fish:
Conditions:
Knockdown Reagents:
Anatomical Terms:
Stage: Day 4
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagents:
Observed In:
Stage: Day 4

Fig. 2 bubblebrain Functions Downstream of Cell Death and Engulfment

(A and B) Dorsal view of blbNY007; Tg(spi1b:Gal4,UAS:GFP); Tg(NBT:DsRed) (A) and blbNY007; Tg(spi1b:Gal4,UAS:GFP); Tg(NBT:DsRed) injected with a Caspase 3 morpholino and treated with Z-VAD to reduce apoptosis (B); representative images quantified in (G).

(C and D) Microglial cells labeled with Tg(spi1:Gal4,UAS:GFP) in 4 dpf blbNY007 (C) and in blbNY007 injected with morpholinos against BAI1 and TIM-4 to reduce engulfment (D); representative images quantified in (H).

(E and F) Representative examples of trunk 4 dpf macrophages in blbNY007 (E) and blbNY007 treated with Camptothecin to increase apoptosis (F). Scale bar 10 μm.

(G) Quantification of (A) and (B). Number of microglia with bubble morphology in blbNY007 (n = 61) and blbNY007 with reduced apoptosis (n = 61). Whiskers: 5th/95th percentile, p value < 0.0001.

(H) Quantification of (C) and (D). Number of microglia with bubble morphology in blbNY007 (n = 50) and blbNY007 injected with Tim4+BAI1 morpholino to reduce engulfment (n = 50). Whiskers: 5th/95th percentile, p value < 0.0001.

(I) Electron micrograph of a representative blbNY007 microglia at 3 dpf. Scale bar, 5 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
DsRed slc37a2t30301/t30301 ; zf148Tg ; zf149Tg control Day 4 brain neuron IFL
slc37a2t30301/t30301 ; zf148Tg ; zf149Tg + MO1-casp3a chemical treatment by environment: Z-Val-Ala-Asp(OMe)-CH2F Day 4 brain neuron IFL
EGFP slc37a2t30301/t30301 ; zf148Tg ; zf149Tg control Day 4 microglial cell IFL
slc37a2t30301/t30301 ; zf148Tg ; zf149Tg + MO1-casp3a chemical treatment by environment: Z-Val-Ala-Asp(OMe)-CH2F Day 4 microglial cell IFL
slc37a2t30301/t30301 ; zf149Tg standard conditions Day 4 microglial cell IFL
slc37a2t30301/t30301 ; zf149Tg + MO1-adgrb1b + MO1-havcr1 standard conditions Day 4 microglial cell IFL
slc37a2t30301/t30301 ; zh901Tg standard conditions Day 4 trunk macrophage IFL
slc37a2t30301/t30301 ; zh901Tg chemical treatment by environment: camptothecin Day 4 trunk macrophage IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
slc37a2t30301/t30301 ; zf148Tg ; zf149Tg control Day 4 microglial cell increased size, abnormal
Day 4 microglial cell phagocytic vesicle increased diameter, abnormal
slc37a2t30301/t30301 ; zf148Tg ; zf149Tg + MO1-casp3a chemical treatment by environment: Z-Val-Ala-Asp(OMe)-CH2F Day 4 brain apoptotic process decreased rate, abnormal
Day 4 microglial cell phagocytic vesicle diameter, ameliorated
Day 4 microglial cell size, ameliorated
slc37a2t30301/t30301 ; zf149Tg standard conditions Day 4 microglial cell increased size, abnormal
Day 4 microglial cell phagocytic vesicle increased diameter, abnormal
slc37a2t30301/t30301 ; zf149Tg + MO1-adgrb1b + MO1-havcr1 standard conditions Day 4 microglial cell phagocytic vesicle diameter, ameliorated
Day 4 microglial cell phagocytosis, engulfment decreased occurrence, abnormal
Day 4 microglial cell size, ameliorated
slc37a2t30301/t30301 ; zh901Tg standard conditions Day 4 trunk macrophage normal size, normal
slc37a2t30301/t30301 ; zh901Tg chemical treatment by environment: camptothecin Day 4 trunk apoptotic process increased rate, abnormal
Day 4 trunk macrophage increased size, 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