Molecular identity and function in transepithelial transport of K(ATP) channels in alveolar epithelial cells.
K(+) channels play a crucial role in epithelia by repolarizing cells and maintaining electrochemical gradient for Na(+) absorption and Cl(-) secretion. In the airway epithelium, the most frequently studied K(+) channels are KvLQT1 and K(Ca). A functional role for K(ATP) channels has been also suggested in the lung, where K(ATP) channel openers activate alveolar clearance and attenuate ischemia-reperfusion injury. However, the molecular identity of this channel is unknown in airway and alveolar epithelial cells (AEC). We adopted an RT-PCR strategy to identify, in AEC, cDNA transcripts for Kir channels (Kir6.1 or 6.2) and sulfonylurea receptors (SUR1, 2A, or 2B) forming K(ATP) channels. Only Kir6.1 and SUR2B were detected in freshly isolated and cultured alveolar cells. To determine the physiological role of K(+) channels in the transepithelial transport of alveolar monolayers, we studied the effect, on total short-circuit currents (I(sc)), of basolateral application of glibenclamide, an inhibitor of K(ATP) channels, as well as clofilium, charybdotoxin, clotrimazole, and iberiotoxin, inhibitors of KvLQT1 and K(Ca) channels, respectively. Interestingly, activity of the three types of K(+) channels was detected, since all tested inhibitors decreased I(sc). Furthermore, these K(+) channel inhibitors reduced amiloride-sensitive Na(+) currents (mediated by ENaC) and completely abolished stimulation of Cl(-) currents by forskolin. Conversely, pinacidil, an activator of K(ATP) channels, increased Na(+) and Cl(-) transepithelial transport by 33-35%. These results suggest the presence, in AEC, of a K(ATP) channel, formed from Kir6.1 and SUR2B subunits, which plays a physiological role, with KvLQT1 and K(Ca) channels, in Na(+) and Cl(-) transepithelial transport.
Leroy
Claudie
C
Centre de recherche, CHUM-Hôtel-Dieu, 3850 St-Urbain, Montréal, Québec H2W 1T7, Canada. emmanuelle.brochiero@umontreal.ca
Dagenais
André
A
Berthiaume
Yves
Y
Brochiero
Emmanuelle
E
D018528:ATP-Binding Cassette Transporters / Q000187:drug effects / Q000502:physiology; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D005576:Colforsin / Q000494:pharmacology; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D005905:Glyburide / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D020110:Pinacidil / Q000494:pharmacology; D015221:Potassium Channels / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D024661:Potassium Channels, Inwardly Rectifying / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011955:Receptors, Drug / Q000187:drug effects / Q000502:physiology; D020545:Respiratory Mucosa / Q000187:drug effects / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D064233:Sulfonylurea Receptors
D018528:ATP-Binding Cassette Transporters; C577509:Abcc8 protein, rat; D017931:DNA Primers; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors; D005576:Colforsin; D020110:Pinacidil; D000584:Amiloride; D004247:DNA; D005905:Glyburide