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Fig. 6. Dispersion of DiD-labelled GFP neurons in the MZ after DiD injection
into the parietal cortex. (A) Schematic of the experimental
paradigm used. DiD-labelled GFP neurons in the MZ were analysed after an
appropriate survival period by preparing flat-mount cortical preparations.
About 70% of DiD-labelled neurons expressed GFP (n=352/485
DiD-labelled cells, 12 brains). The remainder may include Cajal-Retzius cells
(Bielle et al., 2005;
Takiguchi-Hayashi et al.,
2004). (B,C) Distribution of DiD-labelled neurons in
flat-mount preparations one and a half days (E17.0; B) or three days (E18.5;
C) after injection. (B1) Dorsal view of the area around the injection site.
(B2,B3) Higher magnification of the DiD-labelled GFP neurons indicated by
straight lines in B1. These neurons extended an unbranched (B2) or branched
(B3) leading process and a short trailing process. Comparison of fluorescence
in a single optical section demonstrates that this DiD-labelled cell (inset,
magenta) expresses GFP (green). (C1) Dorsal view of the area around the
injection site. (C2-C4) Higher magnification of the DiD-labelled GFP neurons
indicated by straight lines in C1. These neurons showed typical morphology of
migrating neurons in the MZ extending a leading process (C2), occasionally
extending a thick but short process (C3) that appears to elongate toward the
CP (data not shown). Neurons having several long processes, and granular
structure in the trailing process were also observed (C4). (D)
Distribution of DiD-labelled GFP neurons in flat-mount preparations one and a
half (E17.0; black circles) and three days (E18.5; white circles) after
injection. The neurons situated >600 µm or >800 µm from the
injection site were plotted for E17.0 or E18.5 brains, respectively. Pooled
data from two (E17.0) or six (E18.5) independent experiments. The lack of data
medial to the injection site is due to folding of the cortex. OB, olfactory
bulb; R, rostral; C, caudal; L, lateral. Dotted lines, flat-mount outline.
Scale bars: in B1,C1, 200 µm; in B2, B3, 10 µm; in C2-C4, 10 µm.
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