The Na+/H+ antiporter in rat alveolar type II cells and its role in stimulated surfactant secretion.
We used the pH-sensitive fluorescent probe 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) to identify Na+/H+ exchange in freshly isolated rat alveolar type II cells and alveolar type II cells in primary culture. The intracellular pH (pHi) of freshly isolated alveolar type II cells was 7.36 +/- 0.05 (n = 3). When freshly isolated alveolar type II cells were acid loaded with nigericin in sodium-free buffer, the pHi dropped to 6.59 +/- 0.04 and remained low in sodium-free buffer. When acid-loaded cells were subsequently incubated with NaCl, pHi increased in a dose-dependent manner. Amiloride (0.1 mM) inhibited the sodium-induced increase in pHi. When the acid-loaded cells were resuspended in an unbuffered choline chloride solution, the cells secreted H+ in a sodium-dependent and amiloride-inhibitable manner. Alveolar type II cell monolayers, which were cultured for 22 h on glass coverslips and then loaded with BCECF, had a resting pHi of 7.48 +/- 0.05 (n = 4). Nigericin acidified these cultured cells in the absence of sodium and NaCl increased the pHi of these acid loaded cells as observed in freshly isolated cells. Secretagogues of pulmonary surfactant, 12-O-tetradecanoylphorbol 13-acetate (TPA) and terbutaline, did not change pHi. Inhibition of the Na+/H+ antiporter by the addition of amiloride to a Na+ containing medium or the substitution of choline for Na+ did not inhibit stimulated phosphatidylcholine secretion. We conclude that pHi regulation in rat alveolar type II cells is in part mediated by an amiloride-sensitive Na+/H+ antiporter, but this system appears not to be involved in TPA- or terbutaline-induced pulmonary surfactant secretion in primary culture.
Sano
K
K
Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, CO 80206.
Cott
G R
GR
Voelker
D R
DR
Mason
R J
RJ
D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009550:Nigericin / Q000494:pharmacology; D011663:Pulmonary Surfactants / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012965:Sodium Chloride / Q000494:pharmacology; D017923:Sodium-Hydrogen Exchangers; D013726:Terbutaline / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002352:Carrier Proteins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D011663:Pulmonary Surfactants; D017923:Sodium-Hydrogen Exchangers; D012965:Sodium Chloride; D000584:Amiloride; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; D013726:Terbutaline; D013755:Tetradecanoylphorbol Acetate; D009550:Nigericin