Properties of hyperpolarization-activated cation currents (I(h)) were investigated in neurons of juvenile rat entorhinal cortex using the patch-clamp technique. A rat brain slice preparation containing the entorhinal cortex was used for whole-cell recordings of I(h) in pyramidal cells from layer IV and in stellate cells from layer II of the entorhinal cortex. In both stellate and pyramidal cells, I(h) activated at potentials more negative than -60 mV and did not show any time-dependent inactivation. Half-maximal activation of I(h) was achieved at -95.3 mV in pyramidal cells and at -95.0 mV in stellate cells. The channels were permeable for sodium and potassium ions. I(h) of pyramidal and stellate neurons was reduced by about 50% in the presence of 100 microM ZD7288. Extracellularly applied 1 mM Cs(+) decreased I(h) of pyramidal cells by 92%, whereas I(h) of stellate cells was only reduced by 70%. In both pyramidal and stellate neurons, I(h) was not significantly changed during the application of 1 mM Ba(2+). 8-Bromo-c-AMP increased amplitudes of I(h) in stellate cells, while I(h) of pyramidal cells remained unchanged. It is suggested that different types of hyperpolarization-activated cation channels are expressed in pyramidal and stellate cells of the entorhinal cortex.