Generation of stable lipid raft microdomains in the enterocyte brush border by selective endocytic removal of non-raft membrane

PLoS One. 2013 Oct 4;8(10):e76661. doi: 10.1371/journal.pone.0076661. eCollection 2013.

Abstract

The small intestinal brush border has an unusually high proportion of glycolipids which promote the formation of lipid raft microdomains, stabilized by various cross-linking lectins. This unique membrane organization acts to provide physical and chemical stability to the membrane that faces multiple deleterious agents present in the gut lumen, such as bile salts, digestive enzymes of the pancreas, and a plethora of pathogens. In the present work, we studied the constitutive endocytosis from the brush border of cultured jejunal explants of the pig, and the results indicate that this process functions to enrich the contents of lipid raft components in the brush border. The lipophilic fluorescent marker FM, taken up into early endosomes in the terminal web region (TWEEs), was absent from detergent resistant membranes (DRMs), implying an association with non-raft membrane. Furthermore, neither major lipid raft-associated brush border enzymes nor glycolipids were detected by immunofluorescence microscopy in subapical punctae resembling TWEEs. Finally, two model raft lipids, BODIPY-lactosylceramide and BODIPY-GM1, were not endocytosed except when cholera toxin subunit B (CTB) was present. In conclusion, we propose that constitutive, selective endocytic removal of non-raft membrane acts as a sorting mechanism to enrich the brush border contents of lipid raft components, such as glycolipids and the major digestive enzymes. This sorting may be energetically driven by changes in membrane curvature when molecules move from a microvillar surface to an endocytic invagination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Detergents / pharmacology
  • Endocytosis*
  • Enterocytes / metabolism*
  • Enterocytes / ultrastructure
  • Exocytosis
  • Glycolipids / metabolism
  • Intestinal Mucosa / metabolism
  • Jejunum / metabolism
  • Membrane Microdomains / drug effects
  • Membrane Microdomains / metabolism*
  • Microvilli / enzymology
  • Microvilli / metabolism*
  • Microvilli / ultrastructure
  • Organelles / ultrastructure
  • Protein Transport
  • Swine
  • Transcytosis

Substances

  • Detergents
  • Glycolipids

Grants and funding

The study was supported by grants from Augustinus Fonden, Aase og Ejnar Danielsens Fond, Brødrene Hartmanns Fond, Fonden til Lægevidenskabens Fremme, and Hørslev Fonden. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.