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Fig. 9. Notch signaling inhibits both cell-cycle exit and neurogenesis in the
retina. (A-D) BrdU labeling in wild-type (A), add (B), mib
(C) and add; mib mutant retinas. BrdU incorporation is suppressed in
the central region of the add; mib double mutant retina (D,
asterisk). (E-H) ath5 expression in wild-type (E), add (F),
mib (G) and add; mib (H) mutant retinas. ath5
expression is severely inhibited in add mutant retinas, is enhanced
in mib mutant retinas and is partially restored in the add;
mib double mutant retinas (H, asterisk). (I,J) BrdU labeling of
mib mutant injected with the DNA construct pCS2[hsp:EGFP] (I) and
with pCS2[hsp:
47-ß-catenin-GFP] (J). Cells expressing
47-ß-catenin (green, inset in J) are BrdU-positive (red), even in
the central region of mib mutant retinas (J), while cells expressing
EGFP are BrdU negative in the central region of mib mutant retinas
(I). (K,L) BrdU labeling of wild-type retina treated with TSA (K) and
mib mutant retina treated with TSA (L). Treatment with 400 nM TSA
from 10 hpf induces more severe phenotypes than that from 14 hpf shown in
Fig. 5F. (M,N) Plastic sections
of retinas of wild-type retina (M) and wild-type retina expressing NICD (N).
Wild-type retina expressing NICD is folded in the same way as the add
mutant retina (arrowheads). (O,P) Labeling of wild-type (O) and
NICD-expressing (P) retinas with zn5 antibody, which labels retinal ganglion
cells (red). Nuclei were counterlabeled with Sytox Green (green). (Q,R) BrdU
labeling of wild-type (Q) and NICD-expressing retinas (R). In both cases, BrdU
incorporation is not observed in the central retina. Broken white lines show
the interface between the brain and retina. (S,T) Labeling of wild-type (S)
and NICD-expressing retinas (T) with anti-glutamine synthetase antibody, which
labels Müller glia (red). Glial cells differentiate precociously in the
NICD-expressing retina (arrowhead).
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