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Fig. 5. The Pn-1 gene deficiency delays the onset of CGNP
differentiation in the EGL. (A-H) Sections from P10 wild-type
(A,C,E,G) and Pn-1-deficient mice (B,D,F,H) were immunostained for
MATH1 (green; Hoechst, blue). In both groups, MATH1 staining is detected in
the oEGL. The zone of MATH1-positive cells is enlarged in
Pn-1-deficient mice. In addition, the intensity of the MATH1 staining
increases at the cellular level. (I-L) Sections from P10 wild-type and
Pn-1-deficient mice were immunostained for p27 (green; Hoechst, dark
blue; overlap, light blue). The wild-type EGL is divided into two zones: the
oEGL, with few p27-expressing cells, and the iEGL, expressing p27 at high
levels (I,K). In Pn-1-deficient cerebellum, the p27-negative oEGL is
approximately twice the width of the iEGL (J,L). (M,N) Sections
from P10 wild-type and Pn-1-deficient mice injected with BrdU 1 hour
prior to sacrifice were immunostained (BrdU, green; Hoechst, blue). Analysis
of the wild-type oEGL (M) reveals regularly dispersed BrdU-positive CGNPs
(arrowheads), whereas the proliferating CGNPs of Pn-1-deficient mice
are closer to the external pial border (N). (O) The ratio of BrdU
positive versus negative CGNPs is not significantly altered in mutants.
(P) Mutant mice show a significant decrease in the fraction of
p27-labelled CGNPs. *P<0.05 (Student's
t-test). (Q,R) P10 cerebellar sections of
Pn-1+/+ and Pn-1-/- mice immunostained
for GFAP were analyzed by confocal microscopy. Pn-1-/-
Bergmann glia display higher GFAP levels together with an increased thickness
of and larger endfeet (R) in comparison to wild type (Q). iEGL, inner external
granular layer; oEGL, outer external granular layer. Scale bars: 80 µm in B
for A-D; 20 µm in F for E-H; 40 µm in I for I-J and in Q for Q,R; 15
µm in K for K-N and in inset in Q doe insets in Q,R.
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