Involvement of non-selective cationic channels in the generation of pacemaker depolarizations and firing behaviour in cultured frog melanotrophs.

The firing patterns of cultured frog melanotrophs were studied using the patch-clamp technique. In the cell-attached mode, unitary currents were frequently observed as well as biphasic waveforms which were attributed to action potentials 'leaking' through the patch membrane. An inwardly rectifying single-unit current was observed with pipette solutions containing either 100 mM K+ or 100 mM Na+. Under both conditions, these channels displayed an identical I/V relationship, yielding a unitary conductance of 110 pS. The channel opening time was extremely long (50-3000 ms) and single-channel currents showed typical relaxations, which triggered bursts of action currents. In the whole-cell configuration large (2-12 mV) fluctuations in the membrane voltage of current-clamped cells frequently occurred. The deflections appeared to result from single-channel currents. Depolarizing 'events' often led to the discharge of action potentials. Taken together, our data provide evidence for the existence of high-conductance cationic channels in frog pars intermedia cells. These channels may, at least in some cases, be responsible for the generation of pacemaker depolarizations, thereby regulating firing behaviour. It is concluded, that the current traversing a single channel can seriously affect the membrane potential and excitability of frog melanotrophs.