title: Calcium-modulated chloride pathways contribute to chloride flux in murine cystic fibrosis-affected macrophages.
abstract: Cystic fibrosis (CF), a common lethal inherited disorder defined by ion transport abnormalities, chronic infection, and robust inflammation, is the result of mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein, a cAMP-activated chloride (Cl-) channel. Macrophages are reported to have impaired activity in CF. Previous studies suggest that Cl- transport is important for macrophage function; therefore, impaired Cl- secretion may underlie CF macrophage dysfunction. To determine whether alterations in Cl- transport exist in CF macrophages, Cl- efflux was measured using N-[ethoxycarbonylmethyl]- 6-methoxy-quinolinium bromide (MQAE), a fluorescent indicator dye. The contribution of CFTR was assessed by calculating Cl- flux in the presence and absence of cftr(inh)-172. The contribution of calcium (Ca(2+))-modulated Cl- pathways was assessed by examining Cl- flux with varied extracellular Ca(2+) concentrations or after treatment with carbachol or thapsigargin, agents that increase intracellular Ca(2+) levels. Our data demonstrate that CFTR contributed to Cl- efflux only in WT macrophages, while Ca(2+)-mediated pathways contributed to Cl- transport in CF and WT macrophages. Furthermore, CF macrophages demonstrated augmented Cl- efflux with increases in extracellular Ca(2+). Taken together, this suggests that Ca(2+)-mediated Cl- pathways are enhanced in CF macrophages compared with WT macrophages.
authors.affiliation: Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
authors.forename: Ambika
authors.initials: A
authors.lastname: Shenoy
authors.forename: Sascha
authors.initials: S
authors.lastname: Kopic
authors.forename: Michael
authors.initials: M
authors.lastname: Murek
authors.forename: Christina
authors.initials: C
authors.lastname: Caputo
authors.forename: John P
authors.initials: JP
authors.lastname: Geibel
authors.forename: Marie E
authors.initials: ME
authors.lastname: Egan
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002217:Carbachol; D002712:Chlorides / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D005456:Fluorescent Dyes; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D011806:Quinolinium Compounds; D019284:Thapsigargin
chemical_list: D002712:Chlorides; D005456:Fluorescent Dyes; D011806:Quinolinium Compounds; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C085493:N-(ethoxycarbonylmethyl)-6-methoxyquinolinium; D019284:Thapsigargin; D002217:Carbachol; D002118:Calcium