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Rapid and non-genomic reduction of intracellular [Ca²⁺] induced by aldosterone in human bronchial epithelium

Valérie Urbach and Brian J. Harvey *

1. Using a Ca²⁺ imaging system and fura-2 AM (5 µM) we showed that exposure of polarised monolayers of human bronchial epithelial cells (16HBE14o- cell line) to aldosterone produced a fast intracellular [Ca²⁺] ([Ca²⁺]_i) decrease, in 70 % of cells. Exposure to aldosterone (1 nM) reduced the [Ca²⁺]_i by 39 ± 9 nM (n = 282, P < 0.0001) within 10 min, from a basal [Ca²⁺]_i of 131 ± 19 nM (n = 282).
2. The effect of aldosterone on [Ca²⁺]_i was not affected by inhibitors of the classical genomic pathway, cycloheximide (1 µM) or spironolactone (10 µM). The aldosterone-induced [Ca²⁺]_i decrease was inhibited by thapsigargin (1 µM), pertussis toxin (24 h at 200 ng ml^-1), the adenylate cyclase inhibitors 2',3'-dideoxyadenosine (200 µM) and MDL-12,330A hydrochloride (500 µM), and the protein kinase A inhibitor R_P-adenosine 3',5'-cyclic monophosphorothioate (200 µM). In addition, treatment of 16HBE14o- monolayers with aldosterone (1 nM) inhibited by ~30 % the large and transient [Ca²⁺]_i increase induced by apical exposure to uridine triphosphate (UTP, 0.1 mM), a known secretagogue in airway epithelia.
3. Our results demonstrate for the first time that in human bronchial epithelial cells, aldosterone decreases [Ca²⁺]_i levels via a non-genomic mechanism. The hormone-induced changes to [Ca²⁺]_i involve stimulation of thapsigargin-sensitive Ca²⁺-ATPase, via G-protein-, adenylate cyclase- and protein kinase A-coupled signalling pathways.

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