Expression of potassium channels in epithelial cells depends on calcium-activated cell-cell contacts.
Harvesting MDCK cells with trypsin-EDTA reduces potassium currents (IK) to a mere 10%, presumably by hydrolysis of K+ channels, but replating at confluence restores them in 12-18 hr, through a process that requires transcription, translation and exocytic fusion of intracellular membrane vesicles to the plasma membrane (Ponce & Cereijido, 1991; Ponce et al., 1991a). In the present work we find that this restoration of IK also requires cell-cell contacts and the presence of 1.8 mM Ca2+. The role of extracellular Ca2+ may be substituted by 2.0 microM TRH, 10 nM PMA or 200 micrograms/ml DiC8. drugs that stimulate the system of phospholipase C (PLC) and protein kinase C (PKC). Conversely, the recovery of IK triggered by Ca-dependent contacts can be blocked by 110 microM neomycin, 2.0 microM H7, and 250 nM staurosporine, inhibitors of PLC and PKC. These results suggest that the expression of new K+ channels depends on Ca(2+)-activated contacts with neighboring cells and that the information is conveyed through PLC and PKC, a process in keeping with changes in its enzymatic activity and cellular distribution of PKC. Plasma membrane is also reduced and restored upon harvesting and replating, and depends on Ca(2+)-activated contracts. However, the effects of the chemicals tested on IK differ from the ones they elicit on the recovery of plasma membrane, suggesting that cells can independently regulate their population of K+ channels and the surface of their membrane.
Talavera
D
D
Center for Research and Advanced Studies, México, D.F. México.
Ponce
A
A
Fiorentino
R
R
González-Mariscal
L
L
Contreras
R G
RG
Sánchez
S H
SH
García-Villegas
M R
MR
Valdés
J
J
Cereijido
M
M
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002450:Cell Communication; D002460:Cell Line; D002478:Cells, Cultured; D007365:Intercellular Junctions; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D008564:Membrane Potentials; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism
D015221:Potassium Channels; D011493:Protein Kinase C; D010738:Type C Phospholipases; D011188:Potassium; D002118:Calcium