ZFIN ID: ZDB-FIG-180302-19
Seritrakul et al., 2017 - Tet-mediated DNA hydroxymethylation regulates retinal neurogenesis by modulating cell-extrinsic signaling pathways. PLoS Genetics   13:e1006987 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Conditions:
Anatomical Terms:
Stage: Protruding-mouth
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage: Protruding-mouth

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.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP tet3au60/au60 ; zc7Tg ; tet2au59/au59 control Protruding-mouth cranial nerve II IFL
Protruding-mouth photoreceptor cell IFL
Protruding-mouth retinal ganglion cell IFL
tet3au60/au60 ; zc7Tg ; tet2au59/au59 chemical treatment by environment: DAPT Protruding-mouth cranial nerve II IFL
Protruding-mouth green sensitive photoreceptor cell IFL
Protruding-mouth retinal ganglion cell IFL
tet3au60/au60 ; zc7Tg ; tet2au59/au59 chemical treatment by environment: IWR-1-endo Protruding-mouth cranial nerve II IFL
Protruding-mouth green sensitive photoreceptor cell IFL
Protruding-mouth retinal ganglion cell IFL
zc7Tg control Protruding-mouth cranial nerve II IFL
Protruding-mouth retinal ganglion cell IFL
zc7Tg chemical treatment by environment: 6-bromoindirubin-3'-oxime Protruding-mouth cranial nerve II IFL
Protruding-mouth retinal ganglion cell IFL
zc7Tg chemical treatment by environment: DAPT Protruding-mouth cranial nerve II IFL
Protruding-mouth retinal ganglion cell IFL
zc7Tg chemical treatment by environment: IWR-1-endo Protruding-mouth cranial nerve II IFL
Protruding-mouth retinal ganglion cell IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
tet3au60/au60 ; zc7Tg ; tet2au59/au59 control Protruding-mouth cranial nerve II decreased thickness, abnormal
tet3au60/au60 ; zc7Tg ; tet2au59/au59 control Protruding-mouth photoreceptor cell GFP expression decreased amount, abnormal
tet3au60/au60 ; zc7Tg ; tet2au59/au59 control Protruding-mouth retinal ganglion cell decreased amount, abnormal
tet3au60/au60 ; zc7Tg ; tet2au59/au59 chemical treatment by environment: DAPT Protruding-mouth cranial nerve II thickness, ameliorated
Protruding-mouth green sensitive photoreceptor cell GFP expression decreased amount, abnormal
Protruding-mouth retinal ganglion cell amount, ameliorated
tet3au60/au60 ; zc7Tg ; tet2au59/au59 chemical treatment by environment: IWR-1-endo Protruding-mouth cranial nerve II thickness, ameliorated
Protruding-mouth green sensitive photoreceptor cell GFP expression decreased amount, abnormal
Protruding-mouth retinal ganglion cell amount, ameliorated
zc7Tg chemical treatment by environment: 6-bromoindirubin-3'-oxime Protruding-mouth cranial nerve II decreased thickness, abnormal
Protruding-mouth retina layer formation decreased occurrence, abnormal
Protruding-mouth retinal ganglion cell decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.