Cell deformation at the air-liquid interface induces Ca2+-dependent ATP release from lung epithelial cells.
Extracellular nucleotides regulate mucociliary clearance in the airways and surfactant secretion in alveoli. Their release is exquisitely mechanosensitive and may be induced by stretch as well as airflow shear stress acting on lung epithelia. We hypothesized that, in addition, tension forces at the air-liquid interface (ALI) may contribute to mechanosensitive ATP release in the lungs. Local depletion of airway surface liquid, mucins, and surfactants, which normally protect epithelial surfaces, facilitate such release and trigger compensatory mucin and fluid secretion processes. In this study, human bronchial epithelial 16HBE14o(-) and alveolar A549 cells were subjected to tension forces at the ALI by passing an air bubble over the cell monolayer in a flow-through chamber, or by air exposure while tilting the cell culture dish. Such stimulation induced significant ATP release not involving cell lysis, as verified by ethidium bromide staining. Confocal fluorescence microscopy disclosed reversible cell deformation in the monolayer part in contact with the ALI. Fura 2 fluorescence imaging revealed transient intracellular Ca(2+) elevation evoked by the ALI, which did not entail nonspecific Ca(2+) influx from the extracellular space. ATP release was reduced by ∼40 to ∼90% from cells loaded with the Ca(2+) chelator BAPTA-AM and was completely abolished by N-ethylmalemide (1 mM). These experiments demonstrate that in close proximity to the ALI, surface tension forces are transmitted directly on cells, causing their mechanical deformation and Ca(2+)-dependent exocytotic ATP release. Such a signaling mechanism may contribute to the detection of local deficiency of airway surface liquid and surfactants on the lung surface.
Research Centre, Centre hospitalier de l'Université de Montréal (CRCHUM) — Hôtel-Dieu, and Department of Medicine, Université de Montréal, Montréal, Québec, Canada.
Ronaldo
R
Ramsingh
Alexandra
A
Grygorczyk
Anna
A
Solecki
Lalla Siham
LS
Cherkaoui
Yves
Y
Berthiaume
Ryszard
R
Grygorczyk
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000388:Air; D002118:Calcium / Q000378:metabolism*; D048430:Cell Shape / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*; D005033:Ethylmaleimide / Q000494:pharmacology; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D008168:Lung / Q000473:pathology*; D018613:Microscopy, Confocal; D013314:Stress, Mechanical
D000255:Adenosine Triphosphate; D005033:Ethylmaleimide; D002118:Calcium