A, Diagrams showing the location of grids 1 and 2
in relation to ischemic infarct. Grid 1 covers a rectangular area of 280
× 250 μm adjacent to the infarct border. Grid 2 is medial to
grid 1, which is the same size as grid 1. B, Blood
vessels were counted in both grids in 4 serial sections following immunostaining
with mouse-anti-rat RECA-1 antibody. There was a significant increase in the
density of microvessels in stroke rats that received AAV-Netrin-1, compared to
those that received AAV-LacZ in both counting areas. *:
p<0.05; **: p<0.01.
C, Co-localization between Ki67 (green)
and mouse-anti-rat RECA1 in the peri-infarct of a rat receiving AAV-Netrin-1
suggests the presence of newly formed blood vessels (arrowhead). Scale bar: 10
μm. D, Representative photomicrographs
showing the blood vessels stained with mouse-anti-rat RECA-1 antibody in the
peri-infarct cortex of animals receiving AAV-LacZ and AAV-Netrin-1. I: ischemic
core. Scale bar: 100 μm. E, An image of
RECA-1 staining showing the labeled microvessels. The rules for microvessel
counting are described in detail in the material and method section. Scale bar:
15 μm. F, There was a
significant increase in the number of DCX immunoreactive cells detected in the
ipsilateral corpus callosum below the ischemic infarct (*:
p<0.05). However, there was no difference in the number
of DCX cells quantified in either the striatum or peri-infarct cortex.
G, Representative photomicrographs showing
DCX immunoreactive neuroblasts in the ipsilateral striatum, corpus callosum (c.
c.), and peri-infarct cortex at 5 weeks after gene therapy. Scale bar: 50
μm.