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Solitary rod inner segments were obtained by enzymic dissociation of the tiger salamander retina. Ih, an inward current activated by membrane hyperpolarization, was studied using the single-pipette voltage-clamp technique with patch pipettes. In order to investigate Ih in isolation from voltage-dependent potassium and calcium currents, it was necessary to superfuse with a solution containing TEA and cobalt. When the solution in the patch pipette contained 45 mM-KCl and 50 mM-NaCl, the characteristics of Ih were indistinguishable from those previously described with fine-tip micro-electrodes: the reversal potential was near-30 mV and Ih was blocked by extracellular caesium and enhanced by an increase in the extracellular potassium concentration. The increase in Ih observed when the extracellular potassium concentration is raised is due to an increase in conductance and in driving force. Replacement of sodium in the patch pipette with choline caused a 15 mV displacement of the reversal potential for Ih in the depolarized direction. When using sodium-free patch pipettes, replacement of extracellular sodium displaced the reversal potential for Ih to -74 mV, a value in the range of the potassium equilibrium potential in solitary inner segments. Intracellular or intra- and extracellular sodium substitution affected neither the activation range of Ih nor the maximum conductance. From points 3-6 it can be concluded that Ih is carried mainly, if not exclusively, by sodium and potassium and that the channel responsible for Ih is insensitive to modifications of the intra- or extracellular sodium concentration. The results of long-term hyperpolarization, of partial block with caesium and of total sodium substitution are consistent with sodium and potassium permeating the same type of channel.

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