Figure 10.
Figure 10. Involvement of endogenous GABA in the in situ migration of SVZ NG2 cells. (A) Schematic representation of the rat sagittal brain slice showing the location of DiI-stained area (red circle) in the SVZ (gray area). Two arrows indicate the rostral direction (starting from the center of the DiI-stained area and along the longitude axis of the SVZ to the olfactory bulb direction) and dorsal direction (perpendicular with the rostral direction), respectively. The broken blue line crossing the center of the DiI-stained area and forming 45° angles with the two arrows separates the DiI-stained area into the dorsal–rostral (the right side of the broken blue line) and ventral–caudal (the left side of the broken blue line) parts for analyzing cell migration (see below). CC, corpus callosum; CTX, cortex; OB, olfactory bulb. (B, top) Confocal image of the control slice 18 h after live staining with anti-NG2 (green) and DiI crystal (red) showing asymmetric distribution of DiI-labeled cells around the DiI-stained area (outlined by the broken line). Note that more DiI-stained cells, either NG2 positive or NG2 negative, migrated out of the DiI-stained area in the dorsal–rostral side (the right side of the broken line) than those in the ventral–caudal side (the left side of the broken line). Bar, 100 µm. (B, bottom) Higher magnification of the boxed area in the top panel showing that NG2 cells (stained with both DiI and anti-NG2, indicated by arrows) migrated out of the DiI-stained area. Bar, 20 µm. (C, top) Confocal image of the bicuculline-treated slice 18 h after live staining with anti-NG2 (green) and DiI crystal (red) showing symmetric distribution of DiI-labeled cells around the DiI-stained area (outlined by the broken line). Bar, 100 µm. (middle and bottom) Higher magnification of the boxed areas 1 and 2 in the top panel, respectively, showing that double-stained NG2 cells (indicated by arrows) migrated out of the DiI-stained area. Bars, 20 µm. (D) Averaged ratio (number of the migrated NG2 cells in one side/total number of the migrated NG2 cells) of NG2 cells migrating out of the DiI-stained area in the absence (control) or presence of 10 µM bicuculline (Bic), 1 µM TTX, or 10 µM KB-R7943 (KB-R). (E) Averaged total migration distance of NG2 cells (summed migration distance from all the migrated NG2 cells in one side) starting from the edge of the DiI-stained area under various conditions. Data were analyzed from the same brain slices as shown in D. *, P < 0.05; **, P < 0.01 compared between the two groups indicated. n = 23, 13, 9, and 11 slices for control, bicuculline, TTX, and KB-R7943 groups, respectively. Error bars represent mean ± SEM.

Involvement of endogenous GABA in the in situ migration of SVZ NG2 cells. (A) Schematic representation of the rat sagittal brain slice showing the location of DiI-stained area (red circle) in the SVZ (gray area). Two arrows indicate the rostral direction (starting from the center of the DiI-stained area and along the longitude axis of the SVZ to the olfactory bulb direction) and dorsal direction (perpendicular with the rostral direction), respectively. The broken blue line crossing the center of the DiI-stained area and forming 45° angles with the two arrows separates the DiI-stained area into the dorsal–rostral (the right side of the broken blue line) and ventral–caudal (the left side of the broken blue line) parts for analyzing cell migration (see below). CC, corpus callosum; CTX, cortex; OB, olfactory bulb. (B, top) Confocal image of the control slice 18 h after live staining with anti-NG2 (green) and DiI crystal (red) showing asymmetric distribution of DiI-labeled cells around the DiI-stained area (outlined by the broken line). Note that more DiI-stained cells, either NG2 positive or NG2 negative, migrated out of the DiI-stained area in the dorsal–rostral side (the right side of the broken line) than those in the ventral–caudal side (the left side of the broken line). Bar, 100 µm. (B, bottom) Higher magnification of the boxed area in the top panel showing that NG2 cells (stained with both DiI and anti-NG2, indicated by arrows) migrated out of the DiI-stained area. Bar, 20 µm. (C, top) Confocal image of the bicuculline-treated slice 18 h after live staining with anti-NG2 (green) and DiI crystal (red) showing symmetric distribution of DiI-labeled cells around the DiI-stained area (outlined by the broken line). Bar, 100 µm. (middle and bottom) Higher magnification of the boxed areas 1 and 2 in the top panel, respectively, showing that double-stained NG2 cells (indicated by arrows) migrated out of the DiI-stained area. Bars, 20 µm. (D) Averaged ratio (number of the migrated NG2 cells in one side/total number of the migrated NG2 cells) of NG2 cells migrating out of the DiI-stained area in the absence (control) or presence of 10 µM bicuculline (Bic), 1 µM TTX, or 10 µM KB-R7943 (KB-R). (E) Averaged total migration distance of NG2 cells (summed migration distance from all the migrated NG2 cells in one side) starting from the edge of the DiI-stained area under various conditions. Data were analyzed from the same brain slices as shown in D. *, P < 0.05; **, P < 0.01 compared between the two groups indicated. n = 23, 13, 9, and 11 slices for control, bicuculline, TTX, and KB-R7943 groups, respectively. Error bars represent mean ± SEM.

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