IMAGE

Fig. 5

ID
ZDB-IMAGE-120301-35
Genes
Antibodies
Source
Figures for Wan et al., 2012
Image
Figure Caption

Fig. 5

Notch Inhibition Stimulates MG Proliferation and Expands the Zone of Dedifferentiated MG in the Injured Retina via Induction of hb-egfa Gene Expression (A) DAPT treatment of 1016 tuba1a:gfp transgenic fish results in an expansion of the zone of dedifferentiated MG (GFP+/BrdU+) in the injured retina. Scale bar, 50 μm. (B) RT-PCR (gel) and real-time PCR (4 dpi, graph) showing that DAPT induces ascl1a, hb-egfa, and egfr mRNA expression in the injured retina. **p < 0.01 for ascl1a and hb-egfa; *p < 0.05 for egfr. (C) In situ hybridization showing that DAPT expands ascl1a mRNA expression in the injured retina (* marks the injury site). Scale bar, 50 μm. (D) ascl1a:gfp transgenic fish exhibit injury- and Notch-dependent regulation of transgene expression. Scale bar, 50 μm. (E) DAPT stimulates hb-egfa mRNA expression in proliferating MG-derived progenitors (arrows) of the injured retina. Scale bar, 50 μm. (F) MAPK inhibition (PD98059 and SL327) and EGFR inhibition (PD153035) suppress DAPT-dependent expansion of MG proliferation in the injured retina (asterisks in photomicrographs identify injury sites). Graph quantifies data represented in photomicrographs. *p < 0.01. (G) BrdU immunofluorescence on retinal sections reveals BrdU incorporation in the injured retinas of WT and hsp70:gal4;uas:nicd-myc transgenic fish overexpressing NICD. Scale bar, 50 μm. *p < 0.01. (H) In situ hybridization assays for hb-egfa (arrows) and ascl1a mRNA expression in WT and hsp70:gal4;uas:nicd-myc fish overexpressing NICD (* marks the injury site). Scale bar, 50 μm. (I) RT-PCR shows that ascl1a, hb-egfa, and pax6b mRNA expression is inhibited by NICD overexpression. (J) NICD-myc overexpression suppresses HB-EGF-induced cell proliferation in the uninjured retina. BrdU+ cells in HB-EGF-treated retinas overexpressing NICD-myc are generally NICD negative (arrows). Scale bars are 150 μm in low-magnification images and 50 μm in higher-magnification images. Error bars represent SD. ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. See also Figure S6.

Figure Data
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image.

Reprinted from Developmental Cell, 22(2), Wan, J., Ramachandran, R., and Goldman, D., HB-EGF Is Necessary and Sufficient for Müller Glia Dedifferentiation and Retina Regeneration, 334-347, Copyright (2012) with permission from Elsevier. Full text @ Dev. Cell