Receptor-mediated transcytosis of IgA in MDCK cells is via apical recycling endosomes.
Classically, the polymeric immunoglobulin receptor and its ligand, IgA, are thought to be sorted from basolateral early endosomes into transcytotic vesicles that directly fuse with the apical plasma membrane. In contrast, we have found that in MDCK cells IgA is delivered from basolateral endosomes to apical endosomes and only then to the apical cell surface. When internalized from the basolateral surface of MDCK cells IgA is found to accumulate under the apical plasma membrane in a compartment that is accessible to two apically added membrane markers: anti-secretory component Fab fragments, and avidin internalized from the biotinylated apical pole of the cell. This accumulation occurs in the presence of apical trypsin, which prevents internalization of the ligand from the apical cell surface. Using a modification of the diaminobenzidine density-shift assay, we estimate that approximately 80% of basolaterally internalized IgA resides in the apical endosomal compartment. In addition, approximately 50% of basolaterally internalized transferrin, a basolateral recycling protein, has access to this apical endosomal compartment and is efficiently recycled back to the basolateral surface. Microtubules are required for the organization of the apical endosomal compartment and it is dispersed in nocodazole-treated cells. Moreover, this compartment is largely inaccessible to fluid-phase markers added to either pole of the cell, and therefore seems analogous to the recycling endosome described in nonpolarized cells. We propose a model in which transcytosis is not a specialized pathway that uses unique transcytotic vesicles, but rather combines portions of pathways used by non-transcytosing molecules.
Apodaca
G
G
Department of Anatomy, University of California, San Francisco 94143.
Katz
L A
LA
Mostov
K E
KE
D000818:Animals; D001692:Biological Transport; D002451:Cell Compartmentation; D002460:Cell Line; D016764:Cell Polarity; D004285:Dogs; D004705:Endocytosis; D005089:Exocytosis; D005455:Fluorescent Antibody Technique; D007070:Immunoglobulin A / Q000378:metabolism*; D008870:Microtubules / Q000502:physiology; D011971:Receptors, Immunologic; D012635:Secretory Component / Q000378:metabolism*; D014168:Transferrin / Q000378:metabolism; D014404:Tubulin / Q000502:physiology
D007070:Immunoglobulin A; D011971:Receptors, Immunologic; D012635:Secretory Component; D014168:Transferrin; D014404:Tubulin