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Fig. 2. Retinal vascularization correlates with differentiation of retinal
astrocytes. Retinal wholemounts from P5 mice were analyzed for proliferation
(A-G) and stained for PDGFRa (A-C,E), VEGF (G,H) or
GFAP (F,I) mRNA. Retinal astrocytes were visualized by
PDGFRa in situ hybridization (dark signal in A-C,E) and cell
proliferation was assessed by BrdU incorporation (red nuclei in A-C,E-G).
Blood vessels were stained with anti-collagen type IV antibody (green in A,C,
white in H). Proliferation of retinal astrocytes is high in peripheral,
avascular areas (B) but low in central, vascularized areas (C). This was
quantified by counting cells in the avascular (B in panel D) or vascularized
(C in panel D) retina in whole-mount preparations double labeled for
PDGFRa mRNA and BrdU incorporation (data points represent the
mean±s.d. from five different animals). High magnification reveals that
BrdU incorporation in the peripheral retina is limited to retinal astrocytes
identified by PDGFRa mRNA (E), GFAP mRNA (F) and
VEGF mRNA (G) expression. VEGF mRNA (dark signal in H) is
strongly expressed in avascular areas, whereas GFAP mRNA (black in I)
is expressed strongly in the presence of blood vessels. Upper box in A refers
to B, lower box to C. Scale bars: 50 µm.
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