Ischemia Leads to Apoptosis-and Necrosis-like Neuron Death in the Ischemic Rat Hippocampus

Morphological evidence of apoptosis in transient forebrain ischemia is controversial. We therefore investigated the time sequence of apoptosis‐related antigens by immunohistochemistry and correlated it with emerging nuclear patterns of cell death in a model of transient forebrain ischemia in CA1 pyramidal cells of the rat hippocampus. The earliest ischemic changes were found on day 2 and 3, reflected by an upregulation of phospho‐c‐Jun in a proportion of morphologically intact CA1 neurons, which matched the number of neurons that succumbed to ischemia at later time points. At day 3 and later 3 ischemic cell death morphologies became apparent: pyknosis, apoptosis‐like cell death and necrosis‐like cell death, which were confirmed by electron microscopy. Activated caspase‐3 was present in the vast majority of cells with apoptosis‐like morphology as well as in a small subset of cells undergoing necrosis; its expression peaked on days 3 to 4. Silver staining for nucleoli, which are a substrate for caspase‐3, revealed a profound loss of nucleoli in cells with apoptosis‐like morphology, whereas cells with necrosis‐like morphology showed intact nucleoli. Overall, cells with apoptosis‐like morphology and/or caspase‐3 expression represented a minor fraction (