title: Endocytosis of desmosomal plaques depends on intact actin filaments and leads to a nondegradative compartment.
abstract: Epithelial cells in situ can internalize their desmosomes. This can be induced in cell cultures after removal of calcium ions from the cell medium. To study this endocytic process, a nontumorigenic human breast epithelial cell line, HMT-3522, was used. HMT-3522 cells were grown in serum-free, chemically defined medium, containing epidermal growth factor (EGF). Removal of EGF from the medium led to growth arrest and a kind of epithelial differentiation process in which adjacent cells interdigitated and formed more desmosomes than in the proliferating state. Growth-inhibited HMT-3522 cells dissociated following EGTA treatment, the desmosomes divided in a symmetrical fashion, and the desmosomal plaques (half-desmosomes) on the cell surface became internalized. The internalization was independent of clathrin, since immunogold labeling of ultracryosections never showed clathrin on desmosomal plaque-associated membrane domains. Moreover, cytosol acidification, which selectively inhibits endocytosis from clathrin-coated pits, practically blocked the uptake of transferrin, whereas internalization of desmosomal plaques continued. In contrast, actin filaments appeared to be involved in the desmosomal internalization. Thus, depolymerization of actin filaments by cytochalasin D significantly reduced endocytosis of half-desmosomes. Immunogold labeling showed that the vesicles with desmosomal plaques were not enriched in MPR (cation-independent mannose-6-phosphate receptor), cathepsin D or the lysosome-associated membrane protein lamp-1. In addition, the morphology was different. Thus, the endocytic vesicles with desmosomal plaques represent a special compartment, distinct from typical endosomes and lysosomes.
authors.affiliation: Department of Medical Anatomy, Panum Institute, University of Copenhagen, Denmark.
authors.forename: P K
authors.initials: PK
authors.lastname: Holm
authors.forename: S H
authors.initials: SH
authors.lastname: Hansen
authors.forename: K
authors.initials: K
authors.lastname: Sandvig
authors.forename: B
authors.initials: B
authors.lastname: van Deurs
mesh_terms: D000199:Actins / Q000032:analysis / Q000502:physiology*; D001940:Breast / Q000166:cytology / Q000502:physiology / Q000648:ultrastructure; D002460:Cell Line; D002966:Clathrin / Q000502:physiology; D017077:Culture Media, Conditioned / Q000032:analysis / Q000494:pharmacology; D003896:Desmosomes / Q000502:physiology* / Q000648:ultrastructure; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000502:physiology / Q000648:ultrastructure; D004815:Epidermal Growth Factor / Q000032:analysis / Q000494:pharmacology / Q000502:physiology; D004847:Epithelial Cells; D004848:Epithelium / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D007382:Intermediate Filaments / Q000737:chemistry* / Q000502:physiology*; D008247:Lysosomes / Q000502:physiology / Q000648:ultrastructure; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence
chemical_list: D000199:Actins; D002966:Clathrin; D017077:Culture Media, Conditioned; D004815:Epidermal Growth Factor