Definition of distinct compartments in polarized madin-darby canine kidney (MDCK) cells for membrane-volume sorting, polarized sorting and apical recycling

Paul S. Brown, Exing Wang, Benjamin Aroeti, Steven J. Chapin, Keith E. Mostov, Kenneth Dunn

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

Previous studies of fibroblasts have demonstrated that recycling of endocytic receptors occurs through a default mechanism of membrane-volume sorting. Epithelial cells require an additional level of polar membrane sorting, but there are conflicting models of polar sorting, some suggesting that it occurs in early endosomes, others suggesting it occurs in a specialized apical recycling endosome (ARE). The relationship between endocytic sorting to the lysosomal, recycling and transcytotic pathways in polarized cells was addressed by characterizing the endocytic itineraries of LDL, transferrin (Tf) and IgA, respectively, in polarized Madin-Darby canine kidney (MDCK) cells. Quantitative analyses of 3-dimensional images of living and fixed polarized cells demonstrate that endocytic sorting occurs sequentially. Initially internalized into lateral sorting endosomes, Tf and IgA are jointly sorted from LDL into apical and medial recycling endosomes, in a manner consistent with default sorting of membrane from volume. While Tf is recycled to the basolateral membrane from recycling endosomes, IgA is sorted to the ARE prior to apical delivery. Quantifications of the efficiency of sorting of IgA from Tf between the recycling endosomes and the ARE match biochemical measurements of transepithelial protein transport, indicating that all polar sorting occurs in this step. Unlike fibroblasts, rab11 is not associated with Tf recycling compartments in either polarized or glass-grown MDCK cells, rather it is associated with the compartments to which IgA is directed after sorting from Tf. These results complicate a suggested homology between the ARE and the fibroblast perinuclear recycling compartment and provide a framework that justifies previous conflicting models of polarized sorting.

Original languageEnglish
Pages (from-to)124-140
Number of pages17
JournalTraffic
Volume1
Issue number2
StatePublished - 2000

Fingerprint

Madin Darby Canine Kidney Cells
Endosomes
Recycling
Cell Size
Sorting
Cell Membrane
Transferrin
Membranes
Immunoglobulin A
Fibroblasts
Transcytosis
Protein Transport
Glass
Epithelial Cells

Keywords

  • Endocytosis
  • Endosome
  • Epithelia
  • Low density lipoprotein
  • MDCK
  • Polarity
  • Polymeric lg receptor
  • Transcytosis
  • Transferrin

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Structural Biology

Cite this

Definition of distinct compartments in polarized madin-darby canine kidney (MDCK) cells for membrane-volume sorting, polarized sorting and apical recycling. / Brown, Paul S.; Wang, Exing; Aroeti, Benjamin; Chapin, Steven J.; Mostov, Keith E.; Dunn, Kenneth.

In: Traffic, Vol. 1, No. 2, 2000, p. 124-140.

Research output: Contribution to journalArticle

Brown, Paul S. ; Wang, Exing ; Aroeti, Benjamin ; Chapin, Steven J. ; Mostov, Keith E. ; Dunn, Kenneth. / Definition of distinct compartments in polarized madin-darby canine kidney (MDCK) cells for membrane-volume sorting, polarized sorting and apical recycling. In: Traffic. 2000 ; Vol. 1, No. 2. pp. 124-140.
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