Recent studies have shown that prolonged stimulation of afferents of the rat fascia dentata in vivo leads to the development of chronic epileptiform activity of the dentate granule cell region, and degeneration of certain cell types in the adjacent hilus. To investigate the development of dentate hyperexcitability and the selective vulnerability of hilar cells, the hippocampal slice preparation offers an in vitro model in which cellular mechanisms can be examined. We have recorded intracellularly from granule cells and hilar cells in tissue slices from rat before, during, and following sustained stimulation of the major afferent input to the dentate gyrus, the perforant path. Results from intracellular studies in slices were consistent with in vivo studies. Hilar cells were far more sensitive to short-term or prolonged perforant path stimulation than granule cells. At a time when the granule cell population response was not affected by prolonged stimulation, simultaneous recordings from hilar cells and some granule layer interneurons showed that these cells were already depolarized, had very low input resistance, and showed other electrophysiological changes indicative of deterioration. In contrast, granule cells generally hyperpolarized during stimulation and their input resistance increased; no signs of injury were evident in granule cells. Some stimulus-induced changes in the physiological characteristics of granule cells, such as decreased spike frequency adaptation and reduced inhibitory postsynaptic potentials, may contribute to the development of dentate hyperexcitability.