Membrane capacitance and conductance changes parallel mucin secretion in the human airway epithelium.
Measurement of the magnitude and kinetics of exocytosis from intact epithelia has historically been difficult. Using well-differentiated cultures of human bronchial epithelial cells, we describe the use of transepithelial impedance analysis to enable the real-time quantification of mucin secretagogue-induced changes in membrane capacitance (surface area) and conductance. ATPgammaS, UTP, ionomycin, and PMA induced robust increases in total cellular capacitance that were demonstrated to be dominated by a specific increase in apical membrane surface area. The UTP-induced increase in capacitance occurred in parallel with goblet cell emptying and the secretion of mucin and was associated with decreases in apical and basolateral membrane resistances. The magnitude and kinetics of the capacitance increases were dependent on the agonist and the sidedness of the stimulation. The peak increase in capacitance induced by UTP was approximately 30 mucin granule fusions per goblet cell. Secretagogue-induced decreases in apical membrane resistance were independent of exocytosis, although each of the secretagogues induced profound reductions in basolateral membrane resistance. Transepithelial impedance analysis offers the potential to study morphological and conductance changes in cultured human bronchial epithelial cells.
Danahay
Henry
H
Novartis Institutes for Biomedical Research, Horsham, West Sussex, United Kingdom.
Atherton
Hazel C
HC
Jackson
Alan D
AD
Kreindler
James L
JL
Poll
Christopher T
CT
Bridges
Robert J
RJ
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D001980:Bronchi / Q000187:drug effects / Q000378:metabolism*; D002462:Cell Membrane; D002478:Cells, Cultured; D025802:Electric Capacitance; D004553:Electric Conductivity; D005089:Exocytosis; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D009077:Mucins / Q000378:metabolism*; D020545:Respiratory Mucosa / Q000187:drug effects / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014544:Uridine Triphosphate / Q000494:pharmacology
D009077:Mucins; C022571:adenosine 5'-O-(3-thiotriphosphate); D015759:Ionomycin; D000255:Adenosine Triphosphate; D013755:Tetradecanoylphorbol Acetate; D014544:Uridine Triphosphate