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Fig. 3. Inhibition of vascular growth in vivo prevents astrocyte differentiation.
In situ hybridization for PDGFRa (A-C,E,F), VEGF (G) or
GFAP (H) was combined with immunostaining for anti-collagen type IV
(green in A-C,E,F) and anti-BrdU (red in A-C,E,F) on retinal wholemounts.
Exposure of mouse pups to 80% O2 from P0-P8 prevents retinal
vascularization (B,E-H) that normally occurs in control animals (A,C).
Proliferating cells in control animals are limited to vessels (arrowheads, C),
whereas many retinal astrocytes are proliferating in animals lacking retinal
blood vessels (E). However, retinal astrocytes are quiescent in the very
center of the retina (F). Proliferation of retinal astrocytes was quantified
by counting cells in whole-mount retinae (D). Data points represent the
mean±s.d. (from four different animals) of cells counted in normoxic
animals (C in panel D), and in hyperoxic animals in the periphery (E in panel
D) or the center (F in panel D) of the retina. In the central area,
VEGF mRNA expression is low (arrow, G) and GFAP mRNA
expression is high (arrow, H). Box in A refers to C; boxes in B refer to E and
F. Scale bar: 100 µm in A,B,G,H.
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