A specific population of cells located in the hilus of the hippocampal fascia dentata was studied in guinea pig hippocampal slices using standard intracellular recording techniques. Twenty-one such cells were characterized using electrophysiological techniques and were identified morphologically as mossy cells following intracellular injection of the fluorescent dye Lucifer yellow. These cells had a resting membrane potential (mean, -64.6 mV), action potential amplitude (mean, 78.6 mV), action potential duration (mean, 2.2 msec), and time constant (mean, 24.2 msec) similar to those of hippocampal pyramidal cells of area CA3. Rectification seen in their I-V curves, and their ability to fire action potentials in accommodating trains or bursts in response to injected current pulses, were also similar to those of area CA3 pyramidal cells. However, these cells could be distinguished from area CA3 pyramidal cells by their higher input resistance (mean, 97.4 M omega) and higher level of spontaneous activity. The synaptic responses of mossy cells were also different from those of CA3 pyramidal cells. First, mossy cells responded to low levels of stimulation in all areas of the hippocampal slice that were tested, even areas as remote as area CA1. Second, the responses of mossy cells to stimulation consisted primarily of EPSPs. Hyperpolarizing IPSP-like events followed EPSPs in some cells, but the hyperpolarizations were small and monophasic, even after the cell was depolarized with current injection. This response contrasts with the smaller EPSP and the prominent, biphasic IPSP elicited by afferent stimulation of area CA3 pyramidal cells. The physiological and morphological characteristics of these cells suggest that they could play an important role in the integration of electrical activity in the hippocampus.