Intracellular pH changes during neutrophil activation: Na+/H+ antiport.
Activation of the Na+/H+ antiport mechanism was studied in human neutrophils by monitoring intracellular pH with a carboxyfluorescein derivative. N-formyl-methionyl-leucyl-phenylalanine (FMLP) and phospholipase C (PLC) induced biphasic pH changes. Amiloride, which inhibits the antiport, completely blocked alkalinization but enhanced acidification. Polymyxin B, which inhibits protein kinase C, only blocked alkalinization. Activation with phorbol 12-myristate 13-acetate (PMA) led to alkalinization only; this was inhibited by amiloride or polymyxin B. Thus, during polymorphonuclear leukocyte (PMN) activation, intracellular alkalinization appears to be mediated by an amiloride-sensitive Na+/H+ antiport. Antiport activity can also be blocked indirectly by inhibition of protein kinase C activity. Early intracellular acidification does not appear to require kinase activity but is observed when phospholipids are remodeled with PLC. The antiport was also activatable by hypertonic buffered media. This response did not appear to be mediated by protein kinase C because it was unaffected by polymyxin B. Finally, superoxide generation was investigated. It is affected by, but not soley controlled by, either antiport or protein kinase C activity.
D000584:Amiloride / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006982:Hypertonic Solutions; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism*; D011112:Polymyxin B / Q000494:pharmacology; D011493:Protein Kinase C / Q000032:analysis; D011522:Protons; D012964:Sodium / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010738:Type C Phospholipases / Q000494:pharmacology
D006982:Hypertonic Solutions; D011522:Protons; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D000584:Amiloride; D012964:Sodium; D011493:Protein Kinase C; D010738:Type C Phospholipases; D011112:Polymyxin B; D013755:Tetradecanoylphorbol Acetate