Nuclear shrinkage and other markers of neuronal cell death after oxygen-glucose deprivation in rat hippocampal slice cultures

Abstract

Organotypic hippocampal slice cultures are used increasingly in experimental models of neurodegeneration, together with various histological, biochemical and electrophysiological markers of cell death. While functional electrophysiological changes typically occur early, histological changes appear later, with loss of dendritic immunoreactivity for microtubule-associated protein 2 (MAP2) among the earliest. In this study we compared the temporal changes of four different histological markers for neurodegeneration after oxygen-glucose deprivation (OGD) of rat hippocampal slice cultures. Within an observation period of 24 h after OGD, shrinkage of Hoechst 33342 stained neuronal nuclei both occurred before, and was completed faster, than loss of MAP2 staining, which again started earlier and progressed faster towards complete loss than the increase in cellular uptake of propidium iodide and Fluoro-Jade B staining of degenerating neurons. We conclude that shrinkage of Hoechst 33342 stained neuronal nuclei detected by image analysis is an early and easily quantifiable indicator of neuronal degeneration in hippocampal slice cultures.