Influence of TFF3 on neuronal degeneration, cell death, and caspase 3 activity in the ischemic cerebral lesion.

<p>(<b>A</b>) Fluorescence micrographs showing Fluoro-Jade B-labeled degenerating cortical neurons in wild-type and TFF3^-/- mice with cerebral ischemia/reperfusion injury at 24 hrs. No Fluoro-Jade B-labeled cells were found in the cerebrum of sham control mice and the non-injured left hemisphere of the mice with right hemisphere injury. CI: Cerebral ischemia/reperfusion injury. Left: Left hemisphere. Right: Right hemisphere. neg ctrl: Negative control for Fluoro-Jade B staining (without Fluoro-Jade B). Red: MAP2. Green: Fluoro-Jade B (note that the yellowish color represents overlay of Fluoro-Jade B and MAP2 signals). Blue: cell nuclei. Scale bars: 10 µm. (<b>B</b>) TUNEL-labeled cells in the cerebral cortex of wild-type and TFF3^-/- mice with cerebral injury at 24 hrs. The abbreviations are the same as those for panel A, except that neg ctrl is TUNEL-negative controls (specimens incubated with TUNEL reagents excluding fluorophore-conjugated dUTP). No TUNEL-positive cells were found in the cerebrum of sham control mice and the non-injured left hemisphere of the mice with right hemisphere injury. PBS and TFF3: Intravenous administration of PBS or TFF3 immediately after cerebral injury, respectively. Red: MAP2. Green: TUNEL. Blue: cell nuclei. Scale bars: 10 µm. (<b>C</b>) Graphic representation of the density of TUNEL-positive cells in the cerebral cortex of wild-type mice with sham operation (blue) or cerebral ischemia/reperfusion injury (red) and TFF3^-/- mice with cerebral ischemia/reperfusion injury with administration of PBS (purple) or recombinant TFF3 (green). CI: Cerebral ischemia/reperfusion injury. P<0.001 between sham operation (blue) and cerebral ischemia/reperfusion injury (red) in wild-type mice, P<0.001 between wild-type (red) and TFF3^-/- (purple) mice with cerebral ischemia/reperfusion injury, and P<0.001 between cerebrum-injured TFF3^-/- mice with PBS (purple) and recombinant TFF3 (green) at day 1, 3, 5, and 10 days by ANOVA (n = 8). (<b>D</b>) Relative activity of caspase 3 in the ischemic cerebral lesion at 24 hrs, calculated in reference to the relative activity of caspase 3 in the sham control cerebrum.