Journal of Cell Science, Vol 106, Issue 4 1115-1130, Copyright © 1993 by Company of Biologists

JOURNAL ARTICLES

Internalisation of desmosomes and their entry into the endocytic pathway via late endosomes in MDCK cells. Possible mechanisms for the modulation of cell adhesion by desmosomes during development

ID Burdett
Laboratory of Eukaryotic Molecular Genetics, National Institute for Medical Research, Mill Hill, London, UK.

MDCK cells grown in media with normal levels of Ca2+ (approximately 2 mM) contain internalised desmosomes, referred to as desmosome-associated vacuoles (DAVs). The DAVs consist of one to three plaques retained in the plane of a surrounding vacuolar membrane, and their entry into the endocytic pathway has been investigated using HRP, cationized ferritin and BSA/gold in combination with electron microscopy and immunogold labelling of frozen sections. Endocytic tracers supplied from the apical and basolateral surfaces to filter-grown MDCK cells met in a common perinuclear compartment but DAVs were not labelled during short (5-30 minutes) pulses of marker, whether applied apically or basolaterally. Only when the tracers were taken up from the basolateral surface and then chased for periods of 2-18 hours, were DAVs labelled. It is proposed that entry of an endocytic tracer to DAVs occurs by the association of the desmosomal vacuole with late endosomes. Immunolabelling studies with antibodies to desmosomal components (to Dsg, DPI/II), to HRP and to the cation-independent mannose 6-phosphate receptor (MPR), confirmed that Dsg and DPI/II are located within DAVs and late endosomes, but not in early endosomes. Passage of Dsg, but to a lesser extent DPI/II, was detected in MPR- structures (lysosomes). DAV-like structures have also been observed in developing tissues such as mouse kidney. Such engulfment may provide a general mechanism for handling insoluble junctional proteins, particularly where rapid morphogenetic changes are occurring in the pattern of cell-cell adhesion.
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