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1 Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA
2 Department of Cell and Molecular Physiology, University of North Carolina, Chapel Hill, NC, USA; Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA
* To whom correspondence should be addressed. E-mail: cwdavis{at}med.unc.edu.
Studies of regulated mucin secretion from goblet cells in primary cultures of human bronchial epithelial (HBE) cells have suffered, generally, from poor signal:noise ratios, with reported secretory responses of <100% (<1-fold) relative to baseline. Using instead, HBE cells grown as xenografts in the backs of nude mice, UTP (100 µM) stimulated strong mucin secretory responses from isolated, luminally perfused preparations. The peak response (10 min) for 11 control experiments (37 xenografts) was 3.3±0.05-fold relative to baseline, and the time-integrated response (60 min) was 23.4±0.5-fold. Because responses to ATP and UTP were approximately equal, an apical membrane P2Y2-R is suggested. Additionally, ADP activated mucin release from HBE xenografts whereas UDP and 2-methlythio-ADP did not, a pattern of response inconsistent with known purinoceptors. Hence, either a novel receptor to ADP is suggested, or there is significant conversion of ADP to ATP by ecto-adenylate kinase activity. Adenosine and a NO donor were without effect. Consistent with P2Y2-R coupling to phospholipase-C, HBE xenografts responded to ionomycin and PMA; however, they were recalcitrant to forskolin and chlorophenylthio-cAMP, and to 8 Br-cGMP. Hence, human airway goblet cells, like those of other species, appear to be regulated primarily via phospholipase-C pathways, activated particularly by apical membrane P2Y2-R agonists.
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