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Fig. 4. Chemotaxis in MGE-derived cells requires leading process branching.
(A) Schematic diagram of experimental design. (B-D) Migration of
MGE-derived cells in response to mock-transfected (B) or
Nrg1-transfected (C,D) COS cells aggregates cultured in matrigel
matrices for 36 hours in the presence of vehicle solution (B,C) or the ROCK
inhibitor Y27632 (30 µM) (D). COS cells were also transfected with
dsRed to aid their visualization. Broken lines indicate the limits of
the explants before culture. (B'-D') Confocal images
of cells migrating through Sector 1, as defined in the schematic shown in A,
in control (B'), Nrg1 (C') and in Nrg1+Y27632 (30 µM)
(D')-treated explants. Solid and open arrowheads indicate branched and
non-branched interneurons, respectively. (E) Schematic view of the
method used to quantify the orientation of cells (
angle). (F)
Quantification of angle in Sector 1. Bars show mean±s.e.m.
37.02±2.14° (control, n=167 cells from three independent
experiments), 49.65±1.29° (Nrg1, n=520 cells from three
independent experiments) and 38.10±1.41° (Nrg1 +Y27631,
n=381 cells from three independent experiments). t-test,
***P<0.001. (G) Quantification of percentage of
neurons located in Sector 1 with at least two leading process branches. Bars
show mean±s.e.m. 54.85±7.41% (Ctrl, n=161 cells from
three independent experiments), 72.21±4.49% (Nrg1, n=528 cells
from three independent experiments) and 55.73±5.34% (Nrg1+Y27631,
n=317 cells from three independent experiments). t-test,
*P<0.05. Scale bars: 100 µm in B,C,D; 40 µm
B',C',D'.
