Functional assessment of sodium chloride cotransporter NCC mutants in polarized mammalian epithelial cells

American Journal of Physiology. Renal Physiology
L L RosenbaekRobert A Fenton

Abstract

The thiazide-sensitive sodium chloride cotransporter NCC is important for maintaining serum sodium (Na(+)) and, indirectly, serum potassium (K(+)) levels. Functional studies on NCC have used cell lines with native NCC expression, transiently transfected nonpolarized cell lines, or Xenopus laevis oocytes. Here, we developed the use of polarized Madin-Darby canine kidney type I (MDCKI) mammalian epithelial cell lines with tetracycline-inducible human NCC expression to study NCC activity and membrane abundance in the same system. In radiotracer assays, induced cells grown on filters had robust thiazide-sensitive and chloride dependent sodium-22 ((22)Na) uptake from the apical side. To minimize cost and maximize throughput, assays were modified to use cells grown on plastic. On plastic, cells had similar thiazide-sensitive (22)Na uptakes that increased following preincubation of cells in chloride-free solutions. NCC was detected in the plasma membrane, and both membrane abundance and phosphorylation of NCC were increased by incubation in chloride-free solutions. Furthermore, in cells exposed for 15 min to low or high extracellular K(+), the levels of phosphorylated NCC increased and decreased, respectively. To demonstrate that the ...Continue Reading

References

Jun 1, 1984·Pflügers Archiv : European journal of physiology·J M ElaloufC de Rouffignac
Jan 1, 1995·The American Journal of Physiology·J S Adorante
Jul 1, 1996·Kidney International·M D PlotkinS C Hebert
Nov 25, 1998·Proceedings of the National Academy of Sciences of the United States of America·G H KimM A Knepper
Jun 1, 2002·Journal of the American Society of Nephrology : JASN·Joke C De JongRené J M Bindels
Jan 8, 2003·Proceedings of the National Academy of Sciences of the United States of America·Frederick H WilsonRichard P Lifton
Jan 23, 2003·Journal of the American Society of Nephrology : JASN·Robert S HooverSteven C Hebert
Apr 22, 2003·The Journal of Biological Chemistry·Joke C de JongRené J M Bindels
Sep 30, 2003·Journal of the American Society of Nephrology : JASN·Joke C de JongRené J M Bindels
Jan 30, 2004·American Journal of Physiology. Cell Physiology·Otto FröhlichRobert B Gunn
Apr 23, 2004·Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association·Joke C de JongRené J M Bindels
Aug 5, 2006·The Journal of Biological Chemistry·Diana Pacheco-AlvarezGerardo Gamba
May 18, 2007·American Journal of Physiology. Renal Physiology·Monica B SandbergArvid B Maunsbach
Dec 14, 2007·Proceedings of the National Academy of Sciences of the United States of America·Benjamin KoRobert S Hoover
Feb 14, 2008·Journal of Cell Science·Ciaran RichardsonDario R Alessi
Jul 9, 2008·The Journal of Biological Chemistry·Jason D HoffertMark A Knepper
Feb 26, 2009·Proceedings of the National Academy of Sciences of the United States of America·Pedro San-CristobalGerardo Gamba
Apr 30, 2009·The Journal of Biological Chemistry·Arohan R SubramanyaPaul A Welling
Jan 1, 2000·Herzschrittmachertherapie & Elektrophysiologie·G BaumannC Grau
Jul 3, 2009·American Journal of Physiology. Renal Physiology·Volker VallonShinichi Uchida
Apr 16, 2010·American Journal of Physiology. Renal Physiology·Benjamin KoRobert S Hoover
Jul 3, 2010·The Journal of Physiology·Hanne B Moeller, Robert A Fenton
Oct 22, 2010·Canadian Journal of Physiology and Pharmacology·Mark GloverKevin M O'Shaughnessy
Jun 22, 2012·American Journal of Physiology. Renal Physiology·Benjamin KoRobert S Hoover
Oct 5, 2012·American Journal of Physiology. Renal Physiology·Gerardo Gamba
Dec 7, 2013·Pflügers Archiv : European journal of physiology·Arthur D MoesEwout J Hoorn
Mar 20, 2015·American Journal of Physiology. Cell Physiology·Silvana Bazúa-Valenti, Gerardo Gamba
Feb 11, 2016·Annual Review of Physiology·Juliette HadchouelGerardo Gamba
May 7, 2016·Journal of the American Society of Nephrology : JASN·Anne SinningRégine Chambrey
Sep 2, 2016·American Journal of Physiology. Renal Physiology·Marco A Valdez-FloresSabina Jeleń
Jan 6, 2017·Journal of the American Society of Nephrology : JASN·Catherina A CuevasWen-Hui Wang

❮ Previous
Next ❯

Citations

Dec 6, 2018·American Journal of Physiology. Renal Physiology·Anita T Layton
Dec 4, 2019·Physiological Reviews·Ewout J HoornRobert A Fenton
Jun 30, 2019·Journal of the American Society of Nephrology : JASN·Lei ChengRobert A Fenton
Jun 27, 2021·The Journal of Biological Chemistry·Marleen L A KortenoevenRobert A Fenton

❮ Previous
Next ❯

Methods Mentioned

BETA
glycosylation
PCR
Protein Assay
immunoprecipitation
to

Software Mentioned

Graphpad Prism
Image Studio Lite

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

American Journal of Physiology. Renal Physiology
Marco Valdez-FloresSabina Jeleń
Proceedings of the National Academy of Sciences of the United States of America
Kozue YamauchiShinichi Uchida
American Journal of Physiology. Renal Physiology
Bruce WyseDavid H Ellison
© 2022 Meta ULC. All rights reserved