ZFIN ID: ZDB-FIG-190723-1526
Jin et al., 2019 - Dysregulation of Microglial Function Contributes to Neuronal Impairment in Mcoln1a-Deficient Zebrafish. iScience   13:391-401 Full text @ iScience
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Term:
Stage: Day 4
PHENOTYPE:
Fish:
Observed In:
Stage: Day 4

Fig. 3

The Late Endosome and Lysosome Fusion Is Impaired in the biluo Microglia

(A) Fluorescent imaging (green) and LysoTracker staining of 4-dpf sibling, biluo, and Tg(coro1a:mcoln1a);biluo embryos show that the accumulation of acid phagosomes in biluo mutant microglia (middle panel) and morphological defect of microglia in biluo mutants can be rescued by ectopically expressing WT mcoln1a in microglia (lower panel).

(B) Quantification of the number and diameter of LysoTracker-positive phagosomes in the microglia in 4-dpf Tg(coro1a:GFP) siblings (blue dots), Tg(coro1a:GFP);biluo mutants (red squares), and Tg(coro1a:mcoln1a);biluo embryos (green triangles). n(sibling) = 8 microglia from 5 embryos; n(biluo) = 9 microglia from 5 embryos; n(Tg(coro1a:mcoln1a);biluo) = 9 microglia from 5 embryos. **p < 0.01, *p < 0.05, ANOVA. Error bars represent mean ± SD.

(C) Anti-GFP (green), anti-LAMP1, and anti-Lcp1 (blue) triple antibody staining of WT embryos injected H2A:GFP-Rab7. Upper panels represent WT microglia (blue); late endosomes (green) are co-localized with the lysosomes. Lower panels represent mutant microglia (blue); late endosomes (green) are not co-localized with lysosomes.

(D) Quantification of fusion probability of late endosomes and lysosomes in sibling microglia (blue dots) and mutant microglia (red squares). n(sibling) = 6 microglia from 4 embryos, n(biluo) = 6 microglia from 4 embryos. ***p < 0.001. Error bars represent mean ± SD.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP hkz04tTg standard conditions Day 4 optic tectum microglial cell IFL
mcoln1ahkz13/hkz13; hkz04tTg standard conditions Day 4 optic tectum microglial cell IFL
mcoln1ahkz13/hkz13; hkz04tTg; hkz20Tg standard conditions Day 4 optic tectum microglial cell IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
hkz04tTg standard conditions Day 4 late endosome to lysosome transport normal occurrence, normal
Day 4 microglial cell has normal numbers of parts of type microglial cell late endosome, normal
Day 4 microglial cell late endosome normal size, normal
mcoln1ahkz13/hkz13; hkz04tTg standard conditions Day 4 late endosome to lysosome transport decreased occurrence, abnormal
Day 4 microglial cell has extra parts of type microglial cell late endosome, abnormal
Day 4 microglial cell late endosome increased size, abnormal
mcoln1ahkz13/hkz13; hkz04tTg; hkz20Tg standard conditions Day 4 late endosome to lysosome transport occurrence, ameliorated
Day 4 microglial cell has number of microglial cell late endosome, ameliorated
Day 4 microglial cell late endosome size, ameliorated
Acknowledgments:
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