FIGURE

Fig. 5

ID

ZDB-FIG-160308-6

Publication

Shen et al., 2016 - The Rag-Ragulator Complex Regulates Lysosome Function and Phagocytic Flux in Microglia

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Fig. 5

Accumulation of Undigested Neuronal Material in Microglia of In rraga^st77/st77 Mutants

(A–D, G, and H) Confocal images show living larvae bearing transgenes that label neurons Tg(nbt:DsRed) and microglia Tg(mpeg1:EGFP).

(A–E) In WT (A and C) and rraga^st77/st77 mutant (B and D) larvae, arrows indicate microglia that contain neuronal material. In WT larvae, more microglia contain neuronal material at 4 dpf (A and E) than at 6 dpf (C, C′, and E), whereas most microglia in mutants contained neuronal material at both 4 and 6 dpf (B, D, D′, and E). Each point in (E) represents the data from one larva.

(F and G) Frames from time-lapse movies of WT (F) and rraga^st77/st77 (G) mutant larvae starting at 4 dpf, with time points indicated in minutes.

(H) Quantification of the number of neuronal puncta inside microglia in time-lapse movies of WT and rraga^st77/st77 mutant larvae. Each line represents a single microglia (WT, n = 4 cells from three larvae; mutant, n = 4 cells from three larvae).

(I) Immunoblot of whole-animal protein lysates at 5 dpf shows normal LC3-I to LC3-II conversion, but an accumulation of p62 in rraga^st77/st77 mutants. Actin is shown as a loading control.

All larvae shown in (A)–(H) were genotyped by PCR after photography. Larvae analyzed in the immunoblot (I) were genotyped as described in the Experimental Procedures.

Expression Data

Genes:	DsRed EGFP
Antibodies:	Ab2-map1lc3b Ab6-sqstm1
Fish:	WT rraga^st77/st77 gl22Tg; zf148Tg rraga^st77/st77; gl22Tg; zf148Tg
Anatomical Terms:	brain microglial cell brain neuron whole organism
Stage Range:	Day 4 to Day 6

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	rraga^st77/st77 rraga^st77/st77; gl22Tg; zf148Tg
Observed In:	brain apoptotic cell clearance brain engulfment of apoptotic cell microglial cell lysosomal protein catabolic process whole organism
Stage Range:	Day 4 to Day 6

Phenotype Detail

Acknowledgments

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