cAMP-activated Ca2+ signaling is required for CFTR-mediated serous cell fluid secretion in porcine and human airways.
Cystic fibrosis (CF), which is caused by mutations in CFTR, affects many tissues, including the lung. Submucosal gland serous acinar cells are primary sites of fluid secretion and CFTR expression in the lung. Absence of CFTR in these cells may contribute to CF lung pathogenesis by disrupting fluid secretion. Here, we have isolated primary serous acinar cells from wild-type and CFTR-/- pigs and humans without CF to investigate the cellular mechanisms and regulation of fluid secretion by optical imaging. Porcine and human serous cells secrete fluid in response to vasoactive intestinal polypeptide (VIP) and other agents that raise intracellular cAMP levels; here, we have demonstrated that this requires CFTR and a cAMP-dependent rise in intracellular Ca2+ concentration ([Ca2+]i). Importantly, cAMP induced the release of Ca2+ from InsP3-sensitive Ca2+ stores also responsive to cAMP-independent agonists such as cholinergic, histaminergic, and purinergic agonists that stimulate CFTR-independent fluid secretion. This provides two types of synergism that strongly potentiated cAMP-mediated fluid secretion but differed in their CFTR dependencies. First, CFTR-dependent secretion was strongly potentiated by low VIP and carbachol concentrations that individually were unable to stimulate secretion. Second, higher VIP concentrations more strongly potentiated the [Ca2+]i responses, enabling ineffectual levels of cholinergic stimulation to strongly activate CFTR-independent fluid secretion. These results identify important molecular mechanisms of cAMP-dependent secretion, including a requirement for Ca2+ signaling, and suggest new therapeutic approaches to correct defective submucosal gland secretion in CF.
Lee
Robert J
RJ
Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6085, USA.
Foskett
J Kevin
JK
D000328:Adult; D000818:Animals; D000831:Animals, Newborn; D001826:Body Fluids / Q000378:metabolism*; D055745:Bronchioles; D020013:Calcium Signaling; D002217:Carbachol / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D003550:Cystic Fibrosis; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D004847:Epithelial Cells / Q000378:metabolism / Q000502:physiology*; D005088:Exocrine Glands / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000378:metabolism*; D012137:Respiratory System / Q000378:metabolism; D034421:Sus scrofa; D014132:Trachea / Q000378:metabolism
D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D002217:Carbachol; D000242:Cyclic AMP