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Figure 1.
Yap overexpression expands the proliferating cell population in the post-embryonic retina.
(A) Schematic transversal section of a Xenopus tadpole retina (RPE: retinal pigment epithelium; NR: neural retina; ON: optic nerve). Within the CMZ (right panel), retinal stem cells (RSC) reside in the most peripheral margin while actively dividing progenitors (P1) and their post-mitotic progeny (P2) are localized more centrally. (B) In situ hybridization analysis of Yap expression on stage 40 retinal sections. The image on the right is a higher magnification of the CMZ (dashed lines represent the different zones as in a). (C) Immunostaining with anti-YAP antibody on stage 42 retinal sections. YAP labeling is detected in the CMZ as well as in Müller glial cells (arrows). Images on the right are higher magnifications of the CMZ. (D) EdU labeling (3-hr pulse) following in situ hybridization with a Yap probe (dotted line) on stage 40 retinal sections. (E) Lateral views (left panels), head dorsal views (middle panels) and dissected eyes (right panels) of stage 40 tadpoles following two-cell stage microinjection of GFP mRNA as a lineage tracer with either ß-gal (control) or Yap mRNA. The asterisk indicates the injected side. (F) Quantification of dissected eye area. (G–J) TUNEL (G, H; stage 33/34) or EdU incorporation (I, J; 3-hr pulse at stage 40) assays analyzed on retinal sections from tadpoles injected as in (E). Arrows point to TUNEL-positive cells in higher magnifications of the area delineated with dotted line (G). The number of analyzed retinas is indicated in each bar. Data are represented as mean ± SEM. Scale bar = 1 mm in (E) and 40 µm in other panels.
DOI: http://dx.doi.org/10.7554/eLife.08488.003 |
