Neurogenesis in the brain continues throughout life and is promoted by brain insults including
ischemia. There is no critical conclusion, however, about whether proliferated cells acquire neuronal function after
ischemia. Transient global
ischemia was produced by a four-vessel occlusion procedure in rats (n = 54). To label proliferative cells, rats were administrated with a single dose of
5-bromo-2'-deoxyuridine (
BrdU) at 4, 6, 8, 10, 13, or 15 days after
ischemia. Increases in
BrdU-positive cells were detected in the hippocampal dentate gyrus at 5, 7, and 9 days after
ischemia. To determine the phenotype of
BrdU-positive cells,
BrdU was administrated twice daily for 3 consecutive days during 6 to 8 days after
ischemia. A
basic helix-loop-helix transcription factor NeuroD at 7 and 14 days and an immature migrating neuronal marker doublecortin at 14 days after
ischemia were expressed transiently in proliferative cells. These proliferative cells after
ischemia differentiated to the phenotype of neuron at 28 days after
ischemia. Furthermore,
BrdU-positive neurons showed phosphorylation of
extracellular signal-regulated kinase (ERK) by intracerebroventricular injection of
N-methyl-D-aspartate (
NMDA) at 28 and 56 days after
ischemia as seen in surrounding mature neurons. The number of
BrdU-positive neurons, which responded to
NMDA stimulation, increased with time after
ischemia and was greater than that of
sham-operated animals. The present study provides evidence for in vivo ERK phosphorylation in response to
NMDA stimulation of
BrdU-positive neurons in the adult hippocampus after transient forebrain
ischemia.