NDLI: Regulation of intracellular pH in Calu-3 human airway cells

Content Provider	PubMed Central
Author	Inglis, S. K. Finlay, L. Ramminger, S. J. Richard, K. Ward, M. R. Wilson, S. M. Olver, R. E.
Copyright Year	2002
Abstract	The Calu-3 human cell line exhibits features of submucosal gland serous cells and secretes HCO3 −. The aim of this study was to identify the HCO3 − transporters present in these cells by studying their role in the regulation of intracellular pH (pHi). Calu-3 cells were grown on coverslips, loaded with the pH-sensitive fluorescent dye BCECF, and their fluorescence intensity monitored as an indication of pHi. Cells were acidified with NH4Cl (25 mm, 1 min) and pHi recovery recorded. In the absence of HCO3 −, initial recovery was 0.208 ± 0.016 pH units min−1 (n = 37). This was almost abolished by removal of extracellular Na+ and by amiloride (1 mm), consistent with the activity of a Na+-H+ exchanger (NHE). In the presence of HCO3 − and CO2, recovery (0.156 ± 0.018 pH units min−1) was abolished (reduced by 91.8 ± 6.7 %, n = 7) by removal of Na+ but only attenuated (by 63.3 ± 5.8 %, n = 9) by amiloride. 4,4-Dinitrostilbene-2,2-disulfonic acid (DNDS) inhibited recovery by 45.8 ± 5.0 % (n = 7). The amiloride-insensitive recovery was insensitive to changes in membrane potential, as confirmed by direct microelectrode measurements, brought about by changing extracellular [K+] in the presence of either valinomycin or the K+ channel opener 1-EBIO. In addition, forskolin (10 μm), which activates the cystic fibrosis transmembrane conductance regulator Cl− conductance in these cells and depolarises the cell membrane, had no effect on recovery. Removal of extracellular Cl− trebled pHi recovery rates, suggesting that an electroneutral, DNDS-sensitive, Cl−-HCO3 − exchanger together with a NHE may be involved in pHi regulation and HCO3 − secretion in these cells. RT-PCR detected the expression of the electrogenic Na+-HCO3 − cotransporter NBC1 and the Cl−-HCO3 − exchanger (AE2) but not the electroneutral Na+-HCO3 − cotransporter NBCn1.
Related Links	http://dx.doi.org/10.1113/jphysiol.2001.012806
Starting Page	527
File Format	PDF
ISSN	14697793
e-ISSN	14697793
Journal	The Journal of Physiology
Issue Number	Pt 2
Volume Number	538
Language	English
Publisher	Blackwell Science Inc
Publisher Date	2002-01-15
Access Restriction	Open
Rights Holder	Blackwell Science Inc
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Physiology Sports Science

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