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Department of Cell Biology, Lerner Research Institute, and Urological Institute, The Cleveland Clinic, Cleveland, Ohio 44195
The apical- and basolateral-specific
distribution of target soluble N-ethylmaleimide-sensitive
factor attachment protein receptors (t-SNAREs) of the syntaxin family
appear to be critical for polarity in epithelial cells. To test whether
differential SNARE expression and/or subcellular localization may
contribute to the known diversity of trafficking phenotypes of
epithelial cell types in vivo, we have investigated the distribution of
syntaxins 2, 3, and 4 in epithelial cells along the renal tubule.
Syntaxins 3 and 4 are restricted to the apical and basolateral domains,
respectively, in all cell types, indicating that their mutually
exclusive localizations are important for cell polarity. The expression
level of syntaxin 3 is highly variable, depending on the cell type,
suggesting that it is regulated in concert with the cellular
requirement for apical exocytic pathways. While syntaxin 4 localizes
all along the basal and lateral plasma membrane domains in vivo, it is
restricted to the lateral membrane in Madin-Darby canine kidney (MDCK)
cells in two-dimensional monolayer culture. When cultured as cysts in collagen, however, MDCK cells target syntaxin 4 correctly to the basal
and lateral membranes. Unexpectedly, the polarity of syntaxin 2 is
inverted between different tubule cell types, suggesting a role in
establishing plasticity of targeting. The vesicle-associated (v)-SNARE
endobrevin is highly expressed in intercalated cells and colocalizes
with the H+-ATPase in
- but not
-intercalated cells,
suggesting its involvement in H+-ATPase trafficking in the
former cell type. These results suggest that epithelial membrane
trafficking phenotypes in vivo are highly variable and that different
cell types express or localize SNARE proteins differentially as a
mechanism to achieve this variability.
syntaxin; endobrevin; membrane traffic; cell polarity; membrane fusion
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