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Updated: May 1, 2026

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator
Published on: April 5, 2017
Characterization of the Na/HCO3(-) cotransport in human neutrophils
Miriam S Giambelluca1, María C Ciancio, Alejandro Orlowski
1Centro de Investigaciones Cardiovasculares, CCT La Plata, CONICET. Facultad de Ciencias Médicas, Universidad Nacional de La Plata, La Plata, Argentina.
Background:
Bicarbonate transport has crucial roles in regulating intracellular pH (pHi) 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(+)/H(+) 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(+)/HCO3(-) 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 pHi and is modulated by chemotactic agents.
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