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Fig. 2. Branch dynamics during tangential migration. (A) A
telencephalic slice from an E16.5 Gad65-Gfp embryo. GABAergic cells
are observed in green in the different regions in which migration was studied:
subpallium (lateral ganglionic eminence, LGE), cortical subventricular zone
(SVZ) and cortical plate (CP). Owing to the massive accumulation of migrating
neurons in the subpallium, analysis of neurons migrating through the LGE was
performed in slice cultures in which the MGE was electroporated with
Gfp or dsRed (as in Fig.
1). (B-D) Representative images of the morphology of cells
migrating through the LGE (B), SVZ (C) or CP (D).
(B'-D') Quantification of the frequency of angles
form between leading process branches in cells migrating through the LGE
(B'), SVZ (C') or CP (D'). Mean angles are 45.32° (LGE),
35.76° (SVZ) and 51.83° (CP), respectively. Cells inside the highly
packed SVZ generated very small branching angles with high frequency
(C'), whereas neurons in the CP branched at large angles with much more
frequency (D'). The frequency of angles greater than 70° is
significantly higher in the CP than in the LGE or SVZ (
2 test,
***P<0.001). (B'',D'')
Superimposed images of movie frames recreate the path followed by migrating
neurons in the LGE (B'') and CP (D''). Green dots indicate the
position of the nucleus in each nucleokinesis. Diagrams depict directional
changes in these cells. New branches are shown in green, and the chosen branch
is marked with a red arrowhead. Numbers indicate the angle formed by the
branches. ac, anterior commissure; H, hippocampus; MZ, marginal zone; NCx,
neocortex; PCx, piriform cortex; Str, striatum. Scale bars: 500 µm in A; 25
µm in B-D; 20 µm in B'',D''.
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