Characterization of the Na+/HCO3¯ cotransport in human neutrophils

Background: Bicarbonate transport has crucial roles in regulating intracellular pH (pH<SUB>i</SUB>) in a variety of cells. The purpose of this study was to evaluate its participation in the regulation of pHi in resting and stimulated human neutrophils. Methods: Freshly isolated human neu...

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Autores principales: Giambelluca, Miriam Soledad, Ciancio, María Carolina, Orlowski, Alejandro, Gende, Oscar Alfredo, Pouliot, Marc, Aiello, Ernesto Alejandro
Formato: Articulo
Lenguaje:Inglés
Publicado: 2014
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Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/85267
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Sumario:Background: Bicarbonate transport has crucial roles in regulating intracellular pH (pH<SUB>i</SUB>) in a variety of cells. The purpose of this study was to evaluate its participation in the regulation of pHi in resting and stimulated human neutrophils. Methods: Freshly isolated human neutrophils acidified by an ammonium prepulse were used in this study. Results: We demonstrated that resting neutrophils have a bicarbonate transport mechanism that prevents acidification when the Na<SUP>+</SUP>/H<SUP>+</SUP> exchanger is blocked by EIPA. Neutrophils acidified by an ammonium prepulse showed an EIPA-resistant recovery of pHi that was inhibited by the blocker of the anionic transporters SITS or the Na<SUP>+</SUP>/HCO<SUB>3</SUB>¯ cotransporter (NBC) selective inhibitor S0859, and abolished when sodium was removed from the extracellular medium. In western blot and RT-PCR analysis the expression of NBCe2 but not NBCe1 or NBCn1 was detected in neutrophils Acidified neutrophils increased the EIPA-insensitive pHi recovery rate when its activity was stimulated with fMLF/cytochalasin B. This increase in the removal of acid equivalents was insensitive to the blockade of the NADPH oxidase with DPI. Conclusion: It is concluded that neutrophils have an NBC that regulates basal pH<SUB>i</SUB> and is modulated by chemotactic agents.