Hypotonic activation of the myo-inositol transporter SLC5A3 in HEK293 cells probed by cell volumetry, confocal and super-resolution microscopy

PloS One
Joseph AndronicV L Sukhorukov

Abstract

Swelling-activated pathways for myo-inositol, one of the most abundant organic osmolytes in mammalian cells, have not yet been identified. The present study explores the SLC5A3 protein as a possible transporter of myo-inositol in hyponically swollen HEK293 cells. To address this issue, we examined the relationship between the hypotonicity-induced changes in plasma membrane permeability to myo-inositol P ino [m/s] and expression/localization of SLC5A3. P ino values were determined by cell volumetry over a wide tonicity range (100-275 mOsm) in myo-inositol-substituted solutions. While being negligible under mild hypotonicity (200-275 mOsm), P ino grew rapidly at osmolalities below 200 mOsm to reach a maximum of ∼ 3 nm/s at 100-125 mOsm, as indicated by fast cell swelling due to myo-inositol influx. The increase in P ino resulted most likely from the hypotonicity-mediated incorporation of cytosolic SLC5A3 into the plasma membrane, as revealed by confocal fluorescence microscopy of cells expressing EGFP-tagged SLC5A3 and super-resolution imaging of immunostained SLC5A3 by direct stochastic optical reconstruction microscopy (dSTORM). dSTORM in hypotonic cells revealed a surface density of membrane-associated SLC5A3 proteins of 200-2...Continue Reading

References

Sep 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·S UchidaJ S Handler
Apr 9, 1991·Biochimica Et Biophysica Acta·F LangE Tschernko
Dec 1, 1982·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·S GrinsteinA Rothstein
Dec 1, 1982·The Journal of General Physiology·S GrinsteinA Rothstein
Aug 1, 1995·Current Opinion in Cell Biology·H M Kwon, J S Handler
Nov 9, 1995·The New England Journal of Medicine·M L McManusK Strange
Jan 1, 1994·The American Journal of Physiology·H Pasantes-MoralesJ Morán
Jul 1, 1997·The Journal of Membrane Biology·K Kirk
Feb 11, 1998·Physiological Reviews·F LangD Häussinger
Oct 8, 1998·Contributions to Nephrology·R K Kinne
Mar 3, 1999·Proceedings of the National Academy of Sciences of the United States of America·H MiyakawaH M Kwon
Sep 1, 1999·Journal of Neuroscience Research·R E IsaacksM D Norenberg
Jun 3, 2000·American Journal of Physiology. Renal Physiology·S K WooH M Kwon
Dec 23, 2000·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·P R Junankar, K Kirk
Dec 23, 2000·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·R K KinneE Kinne-Saffran
Aug 21, 2001·Pflügers Archiv : European journal of physiology·R FrancoH Pasantes-Morales
Jul 23, 2002·The Journal of Biological Chemistry·Michael J CoadyJean-Yves Lapointe
Apr 11, 2003·Reviews of Physiology, Biochemistry and Pharmacology·F WehnerR K H Kinne
May 17, 2003·Pflügers Archiv : European journal of physiology·Marc Uldry, Bernard Thorens
Jul 4, 2003·American Journal of Physiology. Cell Physiology·Stephen A KempsonMarshall H Montrose
Mar 3, 2004·BioTechniques·Jonathan R Brody, Scott E Kern
Jun 3, 2004·Proceedings of the National Academy of Sciences of the United States of America·Carlos E IrarrazabalJoan D Ferraris
Sep 8, 2004·Analytical Biochemistry·Jonathan R Brody, Scott E Kern
Sep 29, 2004·Pflügers Archiv : European journal of physiology·Stephen A Kempson, Marshall H Montrose
Nov 3, 2004·The Journal of Membrane Biology·R ReussV L Sukhorukov
Jun 20, 2006·Biochemical and Biophysical Research Communications·Vladimir L SukhorukovHeiko Zimmermann
Jun 20, 2006·The Journal of Biological Chemistry·Edith H Y TongBen C B Ko
Jul 18, 2006·The American Journal of Medicine·Herminia Pasantes-MoralesKarina L Tuz
Oct 27, 2007·Journal of the American College of Nutrition·Florian Lang
Jun 27, 2008·The Journal of Biological Chemistry·Min Seong KwonH Moo Kwon
Jul 23, 2008·Angewandte Chemie·Mike HeilemannMarkus Sauer
Jan 8, 2009·Physiological Reviews·Else K HoffmannStine F Pedersen
Jan 9, 2010·Journal of Microscopy·S WolterM Sauer
Mar 9, 2010·Analytical Biochemistry·Isabel Romero-CalvoFermín Sánchez de Medina
May 18, 2011·Biochimica Et Biophysica Acta·Nicole BobakVladimir L Sukhorukov
Jul 2, 2011·Nature Protocols·Sebastian van de LindeMarkus Sauer

❮ Previous
Next ❯

Citations

Dec 10, 2015·Biomicrofluidics·Ryo ShirakashiVladimir L Sukhorukov
Jun 18, 2017·The Journal of Physiological Sciences : JPS·Fumitaka FujitaMakoto Tominaga
Aug 19, 2020·The FEBS Journal·Mattia D PizzagalliGiulio Superti-Furga
May 2, 2017·Disease Markers·Katherine E BeaneyUcleb Consortium
Jul 25, 2020·Journal of Neuroimaging : Official Journal of the American Society of Neuroimaging·Thao T TranKevin S King
Sep 8, 2016·Metallomics : Integrated Biometal Science·Svetlana Lutsenko
Jul 28, 2018·Chemical Reviews·Martin P StewartKlavs F Jensen

❮ Previous
Next ❯

Methods Mentioned

BETA
fluorescence microscopy
PCR
transfection

Software Mentioned

Origin
STORM
rapidSTORM
ImageJ

Related Concepts

Related Feeds

Anxiety Disorders

Discover the latest research on anxiety disorders including agoraphobia, panic disorder, obsessive-compulsive disorder, and post-traumatic stress disorder here.