FIGURE

Fig. 7

ID

ZDB-FIG-180302-19

Publication

Seritrakul et al., 2017 - Tet-mediated DNA hydroxymethylation regulates retinal neurogenesis by modulating cell-extrinsic signaling pathways

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Fig. 7

Fig 7. Tet2 and Tet3 function upstream of the Notch and Wnt pathways during RGC differentiation and morphogenesis.

tet2^-/-;tet3^-/- mutant and sibling embryos carrying isl2b:GFP were exposed to 50μM DAPT, 5μM IWR-1-endo, or 1% DMSO, during neurogenesis (24hpf to 72hpf) and analyzed for isl2b:GFP⁺ RGCs and axons. (A-C, G-H) No significant increases in optic nerve diameter or percentage of isl2b:GFP⁺ RGC per total cells (%RGC) were detected in sibling embryos treated with either 1% DMSO (vehicle), DAPT (Notch inhibitor), or IWR (Wnt inhibitor). (E,G,H) tet2^-/-;tet3^-/- mutants treated with 50μM DAPT showed a significant increase in both the percentage of isl2b:GFP⁺ RGCs per retina and optic nerve diameter, when compared to DMSO-treated controls. (F-H) tet2^-/-;tet3^-/- embryos treated with 5μM IWR also showed a significant increase in the percentage of isl2b:GFP⁺ RGCs per retina and optic nerve diameter. (I-L) Wnt signaling was upregulated by exposing wild-type embryos from 24hpf to 72hpf with 2μM BIO, a GSK3β inhibitor. (I,J) BIO-treated wild-type embryos showed reduced lamination, (K) a decreased percentage of isl2b:GFP⁺ RGCs per retina, and (L) decreased optic nerve diameter relative to DMSO-treated controls, (p = 0.00236 and p<0.0001). All error bars = ± 1 S.D.; n = 5 embryos (A-H) and n = 6 embryos (I-L) per condition analyzed; P-values calculated using two-way ANOVA with multiple comparisons (for G-H) and two-tailed, unpaired t-test (for K-L). Scale bar = 50μm.

Expression Data

Gene:	GFP
Fish:	zc7Tg tet2^au59/au59; tet3^au60/au60; zc7Tg
Conditions:	chemical treatment by environment: 6-bromoindirubin-3'-oxime chemical treatment by environment: DAPT chemical treatment by environment: IWR-1-endo
Anatomical Terms:	cranial nerve II green sensitive photoreceptor cell photoreceptor cell retinal ganglion cell
Stage:	Protruding-mouth

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	zc7Tg tet2^au59/au59; tet3^au60/au60; zc7Tg
Conditions:	chemical treatment by environment: 6-bromoindirubin-3'-oxime chemical treatment by environment: DAPT chemical treatment by environment: IWR-1-endo
Observed In:	cranial nerve II green sensitive photoreceptor cell photoreceptor cell retina layer formation retinal ganglion cell
Stage:	Protruding-mouth

Phenotype Detail

Acknowledgments

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