CL- CURRENTS ACTIVATED BY EXTRACELLULAR NUCLEOTIDES IN HUMAN BRONCHIAL CELLS

The perforated-patch technique was used to study the response of human bronchial cells to extracellular nucleotides. ATP or UTP (100 mu M) e licited a complex response consisting of a large transient membrane cu rrent increase followed by a relatively small sustained level. These t wo phases were characterized by different current kinetics. Throughout the transient phase (2-3 min) the membrane current (I-p) displayed sl ow activation and deactivation kinetics at depolarizing and hyperpolar izing potentials respectively. At steady-state (I-s) the relaxation at hyperpolarizing potential disappeared whereas at positive membrane po tentials the current became slightly deactivating. The I-s amplitude w as dependent on the extracellular Ca2+ concentration, being completely inhibited in Ca2+-free medium. Cell preincubation with the membrane-p ermeable chelating agent BAPTA/AM prevented completely the response to nucleotides, thus suggesting that both I-p and I-s were dependent on intracellular Ca2+. The presence of a hypertonic medium during nucleot ide stimulation abolished I-s leaving I-p unchanged. On the contrary, niflumic acid, a blocker of Ca2+-activated Cl- channels, prevented com pletely I-p without reducing significantly I-s. 1,9-dideoxyforskolin f ully inhibited I-s but also reduced I-p. Replacement of extracellular Cl- with aspartate demonstrated that the currents activated by nucleot ides were Cl- selective. I-p resulted five times more Cl- selective th an I-s with respect to aspartate. Taken together, our results indicate that ATP and UTP activate two types of Cl- currents through a Ca2+-de pendent mechanism.