Am J Physiol Cell Physiol Journal of Applied Physiology
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Am J Physiol Cell Physiol 272: C1295-C1298, 1997;
0363-6143/97 $5.00
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AJP - Cell Physiology, Vol 272, Issue 4 C1295-C1298, Copyright © 1997 by American Physiological Society

ARTICLES

Localization of amiloride-sensitive sodium channels in A6 cells by atomic force microscopy

P. R. Smith, A. L. Bradford, S. Schneider, D. J. Benos and J. P. Geibel
Department of Physiology, Medical College of Pennsylvania and Hahnemann University, Philadelphia 19129, USA.

Atomic force microscopy (AFM) was used for high-resolution imaging of the apical distribution of epithelial Na+ channels in A6 renal epithelial cells. A6 cells grown on coverslips were labeled with antibodies generated against an amiloride-sensitive epithelial Na+ channel complex purified from bovine renal medulla that had been conjugated to 8-nm colloidal gold particles before preparation for AFM. AFM revealed that there was a marked increase in the height of the microvilli in cells labeled with the anti-epithelial Na+ channel antibodies compared with unlabeled cells or cells labeled with rabbit nonimmune immunoglobulin G conjugated to colloidal gold particles. We interpret this apparent increase in microvillar height to be due to anti-epithelial Na+ channel antibody binding to the apical microvilli. These data demonstrate that epithelial Na+ channels are restricted to the apical microvilli in Na+-transporting renal epithelial cells. Furthermore, they demonstrate the applicability of using AFM for high-resolution imaging of the cell surface distribution of epithelial ion channels.


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