Spontaneous and GABA-evoked chloride channels on pituitary intermediate lobe cells and their internal Ca requirements

Abstract

On porcine intermediate lobe (IL) endocrine cells, spontaneously opening chloride channels have been studied and compared to GABA-A activated chloride channels. Elementary currents were recorded mainly from outside-out patches excised from IL cells maintained in culture for 1-4 weeks. Spontaneous inward currents were observed in Cs-loaded cells after replacing Na in the extracellular medium by the impermeant ion choline. This activity, at an internal calcium concentration of 10(-8) M corresponded to a channel for chloride ions with a main conductance level of 26 pS, and substates around 11 pS. The sequence of permeabilities to halides was I greater than Br greater than Cl. These conductance characteristics were common to the GABA-operated channels which also showed a main conductance substate of 23-31 pS. The open time of the 26 pS level mostly encountered in spontaneous activity, was distributed along two modes: one, the most frequent, around 1 ms, and the other around 4 ms. This latter mode was the predominant one observed during GABA and isoguvacine applications but in addition a bursting activity of 19 ms duration was also seen. Specific GABA-A receptor antagonists (bicuculline and SR42641, 1 microM) blocked activity evoked by GABA (1-10 microM), but did not affect spontaneous events. These spontaneous Cl events were only observed in a restricted range of internal Ca concentrations, i.e. between 1 nM and 0.1 microM, and were practically abolished at Cai 1 microM. The GABA-induced activity of Cl channels was also Ca-sensitive, being reduced when Cai reached 1 microM.