title: Basal nucleotide levels, release, and metabolism in normal and cystic fibrosis airways.
abstract: BACKGROUND
  Cystic fibrosis (CF) is a syndrome caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. Despite advances in our understanding of the molecular pathogenesis of CF, the link between CFTR gene mutations and the pathogenesis of CF lung disease remains poorly defined. CFTR has been assigned a number of putative functions that may contribute to innate airway defense, including the regulation of adenosine 5'-triphosphate (ATP) release into the extracellular environment. Because extracellular ATP and uridine 5'-triphosphate (UTP) may regulate airway mucociliary clearance via interaction with luminal P2Y2 receptors, the loss of CFTR-mediated nucleotide release could explain the defect in CF airway defense.
  
  
  METHODS
  We tested the physiologic importance of CFTR-mediated nucleotide release in vivo by directly measuring levels of ATP and UTP in nasal airway surface liquid from normal and CF subjects. Because these basal nucleotide levels reflect the net activities of nucleotide release and metabolic pathways, we also measured constitutive rates of nucleotide release and metabolism on well-differentiated normal and CF airway cultures in vitro. The measurement of ATP release rates were paralleled by in vivo studies employing continuous nasal perfusion in normal and CF subjects. Finally, the regulation of ATP release by isoproterenol and methacholine-stimulated submucosal gland secretion was tested.
  
  
  RESULTS
  These studies revealed that steady-state ATP and UTP levels were similar in normal (470 +/- 131 nM and 37 +/- 7 nM, respectively) and CF (911 +/- 199 nM and 33 +/- 12 nM, respectively) subjects. The rates of both ATP release and metabolism were also similar in normal and CF airway epithelia both in vitro and in vivo. Airway submucosal glands did not secrete nucleotides, but rather, secreted a soluble nucleotidase in response to cholinergic stimuli.
  
  
  CONCLUSIONS
  The concentration of ATP in airway surface liquid is in a range that is relevant for the activation of airway nucleotide receptors. However, despite this finding that suggests endogenous nucleotides may be important for the regulation of mucociliary clearance, our data do not support a role for CFTR in regulating extracellular nucleotide concentrations on airway surfaces.
authors.affiliation: Cystic Fibrosis Research and Treatment Center, University of North Carolina, Chapel Hill 27599, USA. tauras@med.unc.edu
authors.forename: S H
authors.initials: SH
authors.lastname: Donaldson
authors.forename: E R
authors.initials: ER
authors.lastname: Lazarowski
authors.forename: M
authors.initials: M
authors.lastname: Picher
authors.forename: M R
authors.initials: MR
authors.lastname: Knowles
authors.forename: M J
authors.initials: MJ
authors.lastname: Stutts
authors.forename: R C
authors.initials: RC
authors.lastname: Boucher
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000293:Adolescent; D000328:Adult; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D009297:Nasal Mucosa / Q000378:metabolism*
chemical_list: D000255:Adenosine Triphosphate