Gap junction turnover is achieved by the internalization of small endocytic double-membrane vesicles

Mol Biol Cell. 2009 Jul;20(14):3342-52. doi: 10.1091/mbc.e09-04-0288. Epub 2009 May 20.


Double-membrane-spanning gap junction (GJ) channels cluster into two-dimensional arrays, termed plaques, to provide direct cell-to-cell communication. GJ plaques often contain circular, channel-free domains ( approximately 0.05-0.5 mum in diameter) identified >30 y ago and termed nonjunctional membrane (NM) domains. We show, by expressing the GJ protein connexin43 (Cx43) tagged with green fluorescent protein, or the novel photoconvertible fluorescent protein Dendra2, that NM domains appear to be remnants generated by the internalization of small GJ channel clusters that bud over time from central plaque areas. Channel clusters internalized within seconds forming endocytic double-membrane GJ vesicles ( approximately 0.18-0.27 mum in diameter) that were degraded by lysosomal pathways. Surprisingly, NM domains were not repopulated by surrounding channels and instead remained mobile, fused with each other, and were expelled at plaque edges. Quantification of internalized, photoconverted Cx43-Dendra2 vesicles indicated a GJ half-life of 2.6 h that falls within the estimated half-life of 1-5 h reported for GJs. Together with previous publications that revealed continuous accrual of newly synthesized channels along plaque edges and simultaneous removal of channels from plaque centers, our data suggest how the known dynamic channel replenishment of functional GJ plaques can be achieved. Our observations may have implications for the process of endocytic vesicle budding in general.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Endocytosis*
  • Fluorescence
  • Gap Junctions / metabolism*
  • Gap Junctions / ultrastructure
  • HeLa Cells
  • Humans
  • Intracellular Membranes / metabolism*
  • Intracellular Membranes / ultrastructure
  • Membrane Fusion
  • Membrane Lipids / metabolism
  • Membrane Microdomains / metabolism
  • Membrane Microdomains / ultrastructure
  • Models, Biological
  • Rats
  • Transport Vesicles / metabolism*
  • Transport Vesicles / ultrastructure


  • Membrane Lipids