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Fig. 2. Pax6 plays a unique role in each of two spatially distinct subsets of
RPCs in the Pax6flox/flox;
-Cre
OC. The expression of Pax6, VC1.1 (A-H; green and red,
respectively), Crx (I-P), BrdU (Q-V) and syntaxin
(W,X red) were characterized on adjacent sections by antibody
labeling (A-H,Q-X) or in situ hybridization (I-P) in control
(Pax6flox/flox) and mutant
(Pax6flox/flox;-Cre) retinas in the course
of eye development. In the
Pax6flox/flox;-Cre OC, Pax6 was eliminated
from the peripheral regions (B,D,F,H; the Pax6-deficient domain is flanked
with arrowheads). Two spatially distinct populations of Pax6-deficient RPCs
were identified (diagram): the Pax6- cells that are
located in the OC periphery upregulate Crx, whereas the
Pax6- cells located towards the central OC do not
upregulate Crx. The border between the two Pax6-deficient cell types is
indicated with an arrow and a broken line, and their margins are marked with
arrowheads (labeled as region 1 or region 2, respectively). (Y) The
percentage of the Crx-expressing domain (region 1, white bars) and the
Crx- domain (region 2, gray bars) relative to the total
Pax6-deficient area was calculated for E12, E14, E16 OCs (n=4 eyes
for all embryonic stages). (Z) Significant reduction in the percentage
of BrdU+ cells was detected for both Pax6-deficient regions at all
stages of development (**P<0.005 and
*P<0.05 by Student's t-test; n=4 eyes
for all Pax6flox/flox;-Cre retinas and
three eyes for controls). The reduction in the proliferation index was
significantly more extensive in region 1 than in region 2 at E14
(P<0.01 by Student's t-test). inl, inner nuclear layer;
gcl, ganglion cell layer; le, lens; nr, neuroretina; prp, prospective
photoreceptor layer; rpe, retinal pigmented epithelium. Scale bar: 100
µm.
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