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Three-dimensional analysis of post-Golgi carrier exocytosis in epithelial cells

Kreitzer, Geri, Schmoranzer, Jan, Low, Seng Hui, Li, Xin, Gan, Yunbo, Weimbs, Thomas, Simon, Sanford M, Rodriguez-Boulan, Enrique·2003·Nature Cell Biology
DOI 10.1038/ncb917
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Abstract

Targeted delivery of proteins to distinct plasma membrane domains is critical to the development and maintenance of polarity in epithelial cells. We used confocal and time-lapse total internal reflection fluorescence microscopy (TIR-FM) to study changes in localization and exocytic sites of post-Golgi transport intermediates (PGTIs) carrying GFP-tagged apical or basolateral membrane proteins during epithelial polarization. In non-polarized Madin Darby Canine Kidney (MDCK) cells, apical and basolateral PGTIs were present throughout the cytoplasm and were observed to fuse with the basal domain of the plasma membrane. During polarization, apical and basolateral PGTIs were restricted to different regions of the cytoplasm and their fusion with the basal membrane was completely abrogated. Quantitative analysis suggested that basolateral, but not apical, PGTIs fused with the lateral membrane in polarized cells, correlating with the restricted localization of Syntaxins 4 and 3 to lateral and apical membrane domains, respectively. Microtubule disruption induced Syntaxin 3 depolarization and fusion of apical PGTIs with the basal membrane, but affected neither the lateral localization of Syntaxin 4 or Sec6, nor promoted fusion of basolateral PGTIs with the basal membrane.

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  • A crucial mechanism underlying establishment and maintenance of cellular polarity is differential sorting of proteins to distinct plasma membrane domains along the secretory and endosomal pathways.

    “One crucial mechanism underlying establishment and maintenance of cellular polarity is differential sorting of proteins to distinct plasma membrane domains along the secretory and endosomal pathways (Keller et al., 2001;Kreitzer et al., 2003).”

    From SH3TC2, a protein mutant in Charcot–Marie–Tooth neuropathy, links peripheral nerve myelination to endosomal recycling (2010) · cites this paper as Kreitzer et al., 2003

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Metadata

Authors
Kreitzer, G., Schmoranzer, J., Low, S. H., Li, X., Gan, Y., Weimbs, T., Simon, S. M., & Rodriguez-Boulan, E.
Venue
Nature Cell Biology
Publisher
Springer Science and Business Media LLC
Published
2003-01-27
Volume
5
Issue
2
Pages
126-136
ISSN
  • 1465-7392
  • 1476-4679
DOI
10.1038/ncb917
Cited by
196
References
1
Language
en
Licence
www.springer.com/tdm (tdm)

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