title: Abnormal ion content, hydration and granule expansion of the secretory granules from cystic fibrosis airway glandular cells.
abstract: The absence or decreased expression of cystic fibrosis transmembrane conductance regulator (CFTR) induces increased Na(+) absorption and hyperabsorption of the airway surface liquid (ASL) resulting in a dehydrated and hyperviscous ASL. Although the implication of abnormal airway submucosal gland function has been suggested, the ion and water content in the Cystic Fibrosis (CF) glandular secretory granules, before exocytosis, is unknown. We analyzed, in non-CF and CF human airway glandular cell lines (MM-39 and KM4, respectively), the ion content in the secretory granules by electron probe X-ray microanalysis and the water content by quantitative dark field imaging on freeze-dried cryosections. We demonstrated that the ion content (Na(+), Mg(2+), P, S and Cl(-)) is significantly higher and the water content significantly lower in secretory granules from the CF cell line compared to the non-CF cell line. Using videomicroscopy, we observed that the secretory granule expansion was deficient in CF glandular cells. Transfection of CF cells with CFTR cDNA or inhibition of non-CF cells with CFTR(inh)-172, respectively restored or decreased the water content and granule expansion, in parallel with changes in ion content. We hypothesize that the decreased water and increased ion content in glandular secretory granules may contribute to the dehydration and increased viscosity of the ASL in CF.
authors.affiliation: INSERM ERM 203, Laboratoire de Microscopie Electronique Analytique, IFR53, Université de Reims Champagne-Ardenne, 21 rue Clément Ader, 51685 Reims Cedex 2, France.
authors.forename: S
authors.initials: S
authors.lastname: Baconnais
authors.forename: F
authors.initials: F
authors.lastname: Delavoie
authors.forename: J M
authors.initials: JM
authors.lastname: Zahm
authors.forename: M
authors.initials: M
authors.lastname: Milliot
authors.forename: C
authors.initials: C
authors.lastname: Terryn
authors.forename: N
authors.initials: N
authors.lastname: Castillon
authors.forename: V
authors.initials: V
authors.lastname: Banchet
authors.forename: J
authors.initials: J
authors.lastname: Michel
authors.forename: O
authors.initials: O
authors.lastname: Danos
authors.forename: M
authors.initials: M
authors.lastname: Merten
authors.forename: T
authors.initials: T
authors.lastname: Chinet
authors.forename: K
authors.initials: K
authors.lastname: Zierold
authors.forename: N
authors.initials: N
authors.lastname: Bonnet
authors.forename: E
authors.initials: E
authors.lastname: Puchelle
authors.forename: G
authors.initials: G
authors.lastname: Balossier
mesh_terms: D002461:Cell Line, Transformed; D002712:Chlorides / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism / Q000473:pathology; D005088:Exocrine Glands / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008858:Microscopy, Phase-Contrast; D020545:Respiratory Mucosa / Q000378:metabolism* / Q000473:pathology; D022142:Secretory Vesicles / Q000378:metabolism*; D014132:Trachea / Q000378:metabolism* / Q000473:pathology; D014867:Water / Q000378:metabolism*
chemical_list: C505032:CFTR protein, human; D002712:Chlorides; D014867:Water; D019005:Cystic Fibrosis Transmembrane Conductance Regulator