Flow cytometric kinetic assay of the activity of Na+/H+ antiporter in mammalian cells

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
María DolzJuan L Lequerica

Abstract

The Na(+)/H(+) exchanger (NHE) of mammalian cells is an integral membrane protein that extrudes H(+) ion in exchange for extracellular Na(+) and plays a crucial role in the regulation of intracellular pH (pHi). Thus, when pHi is lowered, NHE extrudes protons at a rate depending of pHi that can be expressed as pH units/s. To abolish the activity of other cellular pH-restoring systems, cells were incubated in bicarbonate-free Dulbecco's modified Eagle's medium buffered with HEPES. Flow cytometry was used to determine pHi with 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester or 5-(and-6)-carboxy SNARF-1 acetoxymethyl ester acetate, and the appropriate fluorescence ratios were measured. The calibration of fluorescence ratios versus pHi was established by using ionophore nigericin. The activity of NHE was calculated by a kinetic flow cytometric assay as the slope at time 0 of the best-fit curve of pHi recovery versus time after intracellular acidification with a pulse of exogenous sodium propionate. The kinetic method allowed determination of the pHi-dependent activity of NHE in cell lines and primary cell cultures. NHE activity values were demonstrated to be up to 0.016 pH units/s within the pHi range of ...Continue Reading

References

Jun 11, 1992·American Journal of Respiratory Cell and Molecular Biology·M W RolfeR M Strieter
Jun 1, 1990·The American Journal of Physiology·T I TønnessenS Olsnes
Jan 18, 1989·Biochimica Et Biophysica Acta·S GrinsteinM J Mason
Jan 1, 1986·Annual Review of Physiology·W F Boron
Dec 1, 1993·The Biochemical Journal·L Fliegel, O Fröhlich
Jun 1, 1993·British Journal of Pharmacology·W ScholzB A Schölkens
May 1, 1993·Circulation Research·J I VandenbergA A Grace
Sep 5, 1997·The Journal of Biological Chemistry·J Orlowski, S Grinstein
Apr 18, 1998·Proceedings of the National Academy of Sciences of the United States of America·W C ClaycombN J Izzo
Jan 5, 2000·The Journal of Biological Chemistry·L Counillon, J Pouysségur
Apr 25, 2000·American Journal of Physiology. Heart and Circulatory Physiology·C SchäferH M Piper
Jan 16, 2002·The Journal of Physiology·S K InglisR E Olver
Jan 25, 2002·Annual Review of Pharmacology and Toxicology·L K PutneyD L Barber
Jun 11, 2002·American Journal of Physiology. Cell Physiology·Jingsong ChuMarshall H Montrose
Nov 27, 2002·The Journal of General Physiology·Nicolas Touret, Sergio Grinstein
Feb 22, 2003·Cardiovascular Drug Reviews·W Ross TraceyAngel Guzman-Perez
May 13, 2003·Gut·S TanakaJ D Kaunitz

❮ Previous
Next ❯

Citations

Jul 10, 2007·Journal of Physiology and Biochemistry·J L LequericaB Soria
Aug 29, 2006·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Guadalupe HerreraJosé-Enrique O'Connor
Aug 23, 2016·Nature Communications·Ugur EskiocakSean J Morrison
Jan 4, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Mei K KooShazib Pervaiz
Apr 28, 2020·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Alessandro AliottaLorenzo Alberio

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Stem Cells
Grant A Challen, Melissa H Little
Toxicology in Vitro : an International Journal Published in Association with BIBRA
A Alvarez-BarrientosP Prieto
Cytometry. Part a : the Journal of the International Society for Analytical Cytology
José-Enrique O'ConnorMaría-José Gómez-Lechón
© 2021 Meta ULC. All rights reserved