|
|
||||||||
From the University of North Texas Health Science Center, Department of Anatomy and Cell Biology, Fort Worth, Texas.
Abstract
PURPOSE. To use immunofluorescence and immunogold techniques to identify the catalytic subunits of the Na,K-ATPases of the lens and to determine their location in the cells of the epithelium and cortex of bovine and human lenses.
METHODS. Frozen sections of capsulated and decapsulated bovine and human lenses were prepared, blocked, and treated with affinity-purified polyclonal rabbit antibodies to the Na,K-ATPase catalytic subunit isoforms with subsequent treatment with fluorescein isothiocyanatelabeled goat anti-rabbit IgG and visualization of the fluorescence by light microscopy. An immunogold-labeled goat anti-rabbit IgG was used to detect, by electron microscopy, the binding of the same affinity-purified polyclonal antibodies to thin sections of decapsulated lenses that had been fixed and embedded in Lowicryl K4M. The results were confirmed by staining of western blot analysis of sodium dodecyl sulfatepolyacrylamide gel separations of enriched membrane preparations from bovine and human lenses.
RESULTS. The three common catalytic subunits of the Na,K-ATPases are present in
the plasma membranes of lens epithelium, lens fibers, or both. The data
indicate a polarized distribution of the
1 and
3 catalytic
subunit isoforms in central epithelium. In the cortical fibers, the
2 isoform is present around the interdigitations. The
3 isoform
is found in the interdigitation-free regions of human cortical fibers.
CONCLUSIONS. This unique distribution of Na,K-ATPases precludes the popular pump-leak model for lens monovalent cation homeostasis. The functional significance of the distribution of Na,K-ATPases in the lens epithelium and superficial fibers is currently under investigation.
This article has been cited by other articles:
![]() |
S. Tamiya, W. L. Dean, C. A. Paterson, and N. A. Delamere Regional Distribution of Na,K-ATPase Activity in Porcine Lens Epithelium Invest. Ophthalmol. Vis. Sci., October 1, 2003; 44(10): 4395 - 4399. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. C. Okafor, P. Mukhopadhyay, and N. A. Delamere Studies on Endothelin Release and Na,K Transport in Porcine Lens Invest. Ophthalmol. Vis. Sci., March 1, 2002; 43(3): 790 - 796. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |