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Fig. 1

ID

ZDB-FIG-190724-3

Publication

Villani et al., 2019 - Clearance by Microglia Depends on Packaging of Phagosomes into a Unique Cellular Compartment

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Fig. 1

Characterization of bubblebrain, a Mutant with Bloated Microglia

(A and B) Neutral red staining in a 4 dpf wild-type (A) and blb^NY007 (B) embryos.

(C and D) Microglia labeled with Tg(Apo-E:lynGFP) in a 4 dpf wild-type (C) and blb^NY007 (D) embryo.

(E and F) Microglia cell labeled with Tg(spi1b:Gal4,UAS:GFP) in a 4 dpf wild-type (E) and blb^NY007 (F) embryo. Scale bar, 10 μm.

(G) Quantification of the “bubble” size in blb^NY007 embryos from 2.5 to 4.5 dpf in optic tectum (OT, red plots) and in the hindbrain (HB, orange plots). Whiskers: 5th/95th percentile, p value < 0.0001.

(H) Quantification of apoptotic neurons visualized by Acridine Orange in wild-type (n = 20), blb^NY007 (n = 17) and in wild-type (n = 18) and blb^NY007 (n= 19) injected with pU1 morpholino to deplete microglia in 3.5 dpf embryos; representative images in (J) and (K). Whiskers: 5th/95th percentile, p value < 0.0001.

(I) Quantifications of phagocytic events per cell per hour in wild-type (n = 50; n = 47) and blb^NY007 (n = 61; n = 77) embryos at 3.5 dpf and 4 dpf, respectively. Whiskers: 5th/95th percentile, p value < 0.0001.

(J and K) Dorsal view of a 3.5 dpf wild-type (J) and blb^NY007 (K) embryo injected with pU1 morpholino to deplete microglia and stained with Acridine orange to visualize apoptotic neurons; quantifications in (H).

(L) Microglia cell labeled with Tg(mpeg:GFP-caax) and Tg(NBT:secA5-BFP) to visualize apoptotic neurons in a 4 dpf blb^NY007 embryo.

Expression Data

Genes:	EGFP TagBFP
Fish:	zf147Tg zf149Tg slc37a2^{t30301/t30301}; zf147Tg slc37a2^{t30301/t30301}; zf149Tg slc37a2^{t30301/t30301}; hdb6Tg; zh901Tg
Anatomical Terms:	microglial cell microglial cell phagocytic vesicle lumen microglial cell plasma membrane
Stage:	Day 4

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	slc37a2^{t30301/t30301} WT + MO1-spi1b slc37a2^{t30301/t30301}; zf147Tg slc37a2^{t30301/t30301}; zf149Tg slc37a2^{t30301/t30301} + MO1-spi1b slc37a2^{t30301/t30301}; hdb6Tg; zh901Tg
Knockdown Reagent:	MO1-spi1b
Observed In:	brain brain apoptotic process brain phagocytosis microglial cell microglial cell phagocytic vesicle
Stage Range:	Protruding-mouth to Day 4

Phenotype Detail

Acknowledgments

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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