Figure 6
Hippocampal GCs after 1 day of culture express DCC and UNC5A receptors.
(a)–(f): immunostaining of hippocampal GCs with antibodies for DCC and UNC5A. (a) and (d) DCC (green), (b) and (e) UNC5A (red) and (c) and (f) merged confocal fluorescence images. The arrow in c indicates absence of colocalization, in f the arrow indicates colocalization. (g) DIC image of the GC used for the turning assay using a Netrin-1 (300 ng/ml) gradient. (h) superimposed trajectories of neurite extension during exposure of control (PBS, left) and Netrin-1 gradient (right) for 10 UNC5A-positive (red lines) and 10 UNC5A-negative (black lines) neurons. The pipette position is indicated by the black arrow. Thick line trajectories refer to GCs shown in (i–j) and (k–l) respectively. (i) and (j) Fluorescence image of the same GC shown in g, attracted by Netrin-1, stained for NCAM (green) and UNC5A (red), respectively. (k) and (l) Fluorescence image of the GC repelled by Netrin-1 stained as in i and j. Scale bar: 5 μm.
