spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tousson, A.
Right arrow Articles by Benos, D. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tousson, A.
Right arrow Articles by Benos, D. J.

Journal of Cell Science, Vol 93, Issue 2 349-362, Copyright © 1989 by Company of Biologists

JOURNAL ARTICLES

Immunochemical localization of amiloride-sensitive sodium channels in sodium-transporting epithelia

A Tousson, CD Alley, EJ Sorscher, BR Brinkley and DJ Benos
Department of Cell Biology and Anatomy, University of Alabama, Birmingham 35294.

The localization of amiloride-sensitive Na+ channels in Na+-transporting epithelia was examined using antibodies made against amiloride-binding Na+ channel protein purified from bovine kidney. The distribution of the channel protein was determined in thick frozen sections at the light-microscopic level using indirect immunofluorescence, and at the electron-microscopic level using immunogold labelling. In the cells of both the intact bovine collecting tubule and A6 confluent monolayers, only the luminal or apical-facing surface membranes showed staining. Sodium channel protein was characteristically localized on microvillar domains of the apical plasma membrane. Little or no basolateral membrane staining was evident. Channel protein was also absent from subapical vesicles and tight junctions, and was not found in bovine renal proximal tubules, cultured human secretory sweat coils, non-epithelial Chinese hamster ovary (CHO) cells or human skin fibroblasts. Trypsinization of intact A6 monolayers prior to cell fixation abolished specific staining with antibody. Pretreatment with amiloride protected against this loss of staining. Thus, our probes are specific for amiloride-binding Na+ channel protein, and this channel protein is largely or completely confined to the apical membrane of Na+-transporting epithelia. The level and distribution of specific immunostaining in A6 cells was unchanged by aldosterone treatment, although channel activity, as measured by short-circuit current, increased threefold. This result demonstrates that Na+ channel protein is ever present at the cell surface and exists in both an active and an inactive form. We find no evidence that stimulation of Na+ uptake by aldosterone involves recruitment of new channels from a cytoplasmic pool.


This article has been cited by other articles:


Home page
 Biophys. JHome page
J. Gorelik, Y. Gu, H. A. Spohr, A. I. Shevchuk, M. J. Lab, S. E. Harding, C. R. W. Edwards, M. Whitaker, G. W. J. Moss, D. C. H. Benton, et al.
Ion Channels in Small Cells and Subcellular Structures Can Be Studied with a Smart Patch-Clamp System
Biophys. J., December 1, 2002; 83(6): 3296 - 3303.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
E. Cormet-Boyaka, A. Di, S. Y. Chang, A. P. Naren, A. Tousson, D. J. Nelson, and K. L. Kirk
CFTR chloride channels are regulated by a SNAP-23/syntaxin 1A complex
PNAS, September 17, 2002; 99(19): 12477 - 12482.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
H. Hager, T.-H. Kwon, A. K. Vinnikova, S. Masilamani, H. L. Brooks, J. Frokiaer, M. A. Knepper, and S. Nielsen
Immunocytochemical and immunoelectron microscopic localization of {alpha}-, {beta}-, and {gamma}-ENaC in rat kidney
Am J Physiol Renal Physiol, June 1, 2001; 280(6): F1093 - F1106.
[Abstract] [Full Text] [PDF]

Home page
 J. Histochem. Cytochem.Home page
O. Kretz, P. Barbry, R. Bock, and B. Lindemann
Differential Expression of RNA and Protein of the Three Pore-forming Subunits of the Amiloride-sensitive Epithelial Sodium Channel in Taste Buds of the Rat
J. Histochem. Cytochem., January 1, 1999; 47(1): 51 - 64.
[Abstract] [Full Text]

Home page
 Ann. N. Y. Acad. Sci.Home page
B. LINDEMANN, P. BARBRY, O. KRETZ, and R. BOCK
Occurrence of ENaC Subunit mRNA and Immunocytochemistry of the Channel Subunits in Taste Buds of the Rat Vallate Papilla
Ann. N.Y. Acad. Sci., November 30, 1998; 855(1): 116 - 127.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
R. G. Morris, A. Tousson, D. J. Benos, and J. A. Schafer
Microtubule disruption inhibits AVT-stimulated Cl- secretion but not Na+ reabsorption in A6 cells
Am J Physiol Renal Physiol, February 1, 1998; 274(2): F300 - F314.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
C Roudier-Pujol, A Rochat, B Escoubet, E Eugene, Y Barrandon, J. Bonvalet, and N Farman
Differential expression of epithelial sodium channel subunit mRNAs in rat skin
J. Cell Sci., January 2, 1996; 109(2): 379 - 385.
[Abstract] [PDF]

Home page
 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
A. Lazrak, U. Thome, C. Myles, J. Ware, L. Chen, C. J. Venglarik, and S. Matalon
Alveolar Epithelial Ion and Fluid Transport: cAMP regulation of Cl- and HCO3- secretion across rat fetal distal lung epithelial cells
Am J Physiol Lung Cell Mol Physiol, April 1, 2002; 282(4): L650 - L658.
[Abstract] [Full Text] [PDF]


© The Company of Biologists Ltd 1989