Regulation of Na+/H+ and Cl-/HCO3- antiports in Vero cells.
The effect of serum, phorbol-12-myristate-13-acetate (TPA), and forskolin on the activity Na+/H+ antiport and the Na(+)-coupled and Na(+)-independent Cl-/HCO3- antiport was studied in Vero cells by measuring 22Na+ and 36Cl- fluxes and changes in cytosolic pH (pHi). The Na(+)-independent Cl-/HCO3- antiport, which acts as an acidifying mechanism, is strongly pH-sensitive. In serum-starved cells it is activated at alkaline cytosolic pH, with a half-maximal activity at pHi approximately 7.20. Incubation with serum increased the activity of the Na(+)-independent Cl-/HCO3- antiport at pHi values from 6.8 to 7.2. Thus serum appeared to alter the pHi sensitivity of this antiporter such that the threshold value for activation of the antiport was shifted to a more acidic value. Na+/H+ antiport was somewhat stimulated initially by addition of serum, but further incubation with serum (greater than 45 min) decreased its activity. The activity of the Na(+)-coupled Cl-/HCO3- antiport, which is the major alkalinizing antiport in Vero cells, was not altered by short-term incubation with serum (less than 10 min) but decreased after prolonged incubation (greater than 45 min). Our findings with TPA and forskolin indicate that the effect of serum is partly mediated by the protein kinase C pathway, whereas the cyclic adenosine monophosphate pathway does not appear to play an important role. The net effect of serum on the pHi-regulating antiports was a slight decrease in intracellular pH.
Tønnessen
T I
TI
Institute for Cancer Research, Norwegian Radium Hospital, Montebello, Oslo.
Aas
A T
AT
Ludt
J
J
Blomhoff
H K
HK
Olsnes
S
S
D000818:Animals; D001639:Bicarbonates / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D003470:Culture Media; D003600:Cytosol / Q000502:physiology; D006133:Growth Substances / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D017923:Sodium-Hydrogen Exchangers; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014709:Vero Cells / Q000378:metabolism*
D001639:Bicarbonates; D002352:Carrier Proteins; D002712:Chlorides; D003470:Culture Media; D006133:Growth Substances; D017923:Sodium-Hydrogen Exchangers; D005576:Colforsin; D013755:Tetradecanoylphorbol Acetate