title: Internal alkalinization by reversal of anion exchange in human neutrophils: regulation of transport by pH.
abstract: When steady-state human neutrophils bathed in 148 mM Cl- are transferred to a Cl(-)-free medium containing 0.5 mM HCO3- and 148 mM glucuronate or aspartate as nominally inert replacement ions, there is a rapid efflux of 36Cl- from the cells. The accelerated loss of Cl- is accompanied by an intracellular alkalinization of 0.7-0.9 pH units. Both the Cl- efflux and intracellular pH (pHi) transient are dependent on extracellular HCO3- and are sensitive to inhibition by SITS and alpha-cyano-4-hydroxycinnamate, which block anion exchange, thereby indicating that these processes are due to the countertransport of internal Cl- for external HCO3-. Rate of anion exchange is strongly influenced by pH, which functions to regulate carrier activity; alkalinization stimulates the transport velocity, whereas acidification inhibits it. The relationship to pHi follows a Hill equation with pK' approximately 7.40 and Hill coefficient of 3.3, thereby suggesting that approximately 3 HCO3- may be required to bind to the modifier site. Neutrophils placed in glucuronate medium progressively shrink during the first 7.5 min of incubation due to the net loss of osmotically active particles through Cl(-)-HCO3- exchange. However, between 7.5 and 30 min, cells regain their normal cell size. This volume recovery phase correlates with the time course of 22Na+ and [14C]glucuronate influxes, whose kinetics can be dissociated from that of anion exchange. Uptake of glucuronate is largely Na+ dependent (whereas Cl(-)-HCO3- exchange is not), is resistant to amiloride, and can be blocked by furosemide, which suggests that glucuronate probably enters via a volume-activated pathway such as Na+ + glucuronate cotransport.
authors.affiliation: Department of Medicine, Veterans Administration Medical Center, St. Louis, Missouri.
authors.forename: L
authors.initials: L
authors.lastname: Simchowitz
authors.forename: A O
authors.initials: AO
authors.lastname: Davis
mesh_terms: D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000494:pharmacology; D000584:Amiloride / Q000494:pharmacology; D001224:Aspartic Acid / Q000097:blood; D001639:Bicarbonates / Q000097:blood; D001692:Biological Transport; D002712:Chlorides / Q000097:blood*; D005665:Furosemide / Q000494:pharmacology; D005965:Glucuronates / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000502:physiology*
chemical_list: D001639:Bicarbonates; D002712:Chlorides; D005965:Glucuronates; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D001224:Aspartic Acid; D000584:Amiloride; D005665:Furosemide