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Vol. 10, Issue 1, 47-61, January 1999
and§
Recent evidence suggests that apical and basolateral
endocytic pathways in epithelia converge in an apically located,
pericentriolar endosomal compartment termed the apical recycling
endosome. In this compartment, apically and basolaterally internalized
membrane constituents are thought to be sorted for recycling back to
their site of origin or for transcytosis to the opposite plasma
membrane domain. We report here that in the epithelial cell line
Madin-Darby Canine Kidney (MDCK), antibodies to Rab11a label an
apical pericentriolar endosomal compartment that is dependent on intact
microtubules for its integrity. Furthermore, this compartment is
accessible to a membrane-bound marker (dimeric immunoglobulin A
[IgA]) internalized from either the apical or basolateral pole,
functionally defining it as the apical recycling endosome. We have also
examined the role of a closely related epithelial-specific Rab, Rab25,
in the regulation of membrane recycling and transcytosis in MDCK cells. When cDNA encoding Rab25 was transfected into MDCK cells, the protein
colocalized with Rab11a in subapical vesicles. Rab25 transfection also
altered the distribution of Rab11a, causing the coalescence of
immunoreactivity into multiple denser vesicular structures not
associated with the centrosome. Nevertheless, nocodazole still dispersed these vesicles, and dimeric IgA internalized from either the
apical or basolateral membrane was detected in endosomes labeled with
antibodies to both Rab11a and Rab25. Overexpression of Rab25 decreased
the rate of IgA transcytosis and of apical, but not basolateral,
recycling of internalized ligand. Conversely, expression of the
dominant-negative Rab25T26N did not alter either apical recycling or
transcytosis. These results indicate that both Rab11a and Rab25
associate with the apical recycling system of epithelial cells and
suggest that Rab25 may selectively regulate the apical recycling and/or
transcytotic pathways.
Institute for Molecular Medicine and
Genetics, Departments of Medicine, Surgery, Cellular Biology and
Anatomy, Medical College of Georgia and the Augusta Veterans Affairs
Medical Center, Augusta, Georgia 30912;
*Pediatric Gastroenterology
Unit, Massachusetts General Hospital East, and Program in Biological
and Biomedical Sciences, Harvard University Medical School,
Charlestown, Massachusetts 02129; and
Department of
Anatomy, University of California, San Francisco, California 94143
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