Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10769971 | Biochemical and Biophysical Research Communications | 2005 | 9 Pages |
Abstract
Using the whole-cell patch-clamp technique, we identified an amiloride (AMI)-sensitive Na+ current in cystic fibrosis cells, JME/CF15, growing in standard medium. The reversal potential of this current depended on Na+ concentrations and the cation selectivity was much higher for Na+ than for K+, indicating that the current is through ENaC channels. In contrast, cells from EGF-containing medium lacked AMI-sensitive Na+ currents. In permeabilized cells growing in EGF-containing medium, αENaC was mainly detected in a perinuclear region, while in cells from standard medium it was distributed over the cell body. Western-blot analysis showed that in standard medium cells expressed fast-migrating EndoH-insensitive and slow-migrating EndoH-sensitive αENaC fractions, while in cells growing in the presence of EGF, αENaC was only detected as the fast-migrating EndoH-insensitive fraction. Long-term incubation of cells with EGF resulted in an increased basal Ca2+ level, [Ca2+]i. A similar increase of [Ca2+]i was also observed in the presence of 2 μM thapsigargin, resulting in inhibition of ENaC function. Thus, in JME/CF15 cells inhibition of the ENaC function by chronic incubation with EGF is a Ca2+-mediated process that affects trafficking and surface expression of ENaC channels.
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Authors
Lishuang Cao, Grzegorz Owsianik, Frédéric Becq, Bernd Nilius,