CFTR and tight junctions in cultured bronchial epithelial cells
2010 (English)In: Experimental and molecular pathology (Print), ISSN 0014-4800, E-ISSN 1096-0945, Vol. 88, no 1, 118-127 p.Article in journal (Refereed) Published
Airway epithelial salt and water transport takes place through paracellular and transcellular pathways. This transport depends critically on the epithelial sodium channel (ENaC) and the cystic fibrosis transmembrane conductance regulator (CFTR), operating in concert with the paracellular pathway through the tight junctions (TJs). Normal (16HBE14o-), cystic fibrosis (CFBE41o-), and corrected CFBE41o- (CFBE41o pCep4) airway epithelial cell lines were cultured under isotonic conditions. Transepithelial electrical resistance (TEER) was measured as indicator of the tightness of the cultures. Morphology was investigated by immunofluorescence and paracellular permeability by lanthanum nitrate or [14C] mannitol as permeability markers. CFBE41o pCep4 cells developed lower TEER than CFBE41o- cells. Addition of a specific inhibitor of CFTR (CFTRinh-172) to 16HBE14o- and CFBE pCep4 cells resulted in a time-dependent increase in TEER whereas stimulation of CFTR by IBMX and forskolin caused a decrease. Permeability to lanthanum and [14C] mannitol was lower in 16HBE14o- cells exposed to CFTRinh-172 and in CFBE41o- cells compared to untreated 16HBE14o- and CFBE41o pCep4 cells, respectively. 16HBE14o- cells exposed to IBMX and forskolin showed higher permeability to lanthanum but lower permeability to [14C] mannitol compared to control. Immunofluorescence revealed a disorganisation of F-actin and a-tubulin in 16HBE14o- cells exposed to CFTRinh-172, which was not seen in untreated cultures. A higher degree of disorganised F-actin and a-tubulin was also seen in CFBE41o- cells compared to CFBE41o- pCep4 cells. Changes in F-actin and a-tubulin in 16HBE14o- cells exposed to IBMX and forskolin were also seen, although these were less apparent. These results suggest the possibility of an interaction between the activity of CFTR and the TJ protein complex, probably via the cytoskeleton.
Place, publisher, year, edition, pages
2010. Vol. 88, no 1, 118-127 p.
CFTR, transepithelial electrical resistance, tight junctions, cytoskeleton
Medical and Health Sciences
Research subject Biology with specialization in Molecular Cell Biology; Molecular Medicine
IdentifiersURN: urn:nbn:se:uu:diva-99427DOI: 10.1016/j.yexmp.2009.09.018ISI: 000274500100017PubMedID: 19818767OAI: oai:DiVA.org:uu-99427DiVA: diva2:208571