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The Journal of Membrane Biology

, Volume 245, Issue 7, pp 411–422 | Cite as

Cx43 Associates with Nav1.5 in the Cardiomyocyte Perinexus

  • J. Matthew Rhett
  • Emily L. Ongstad
  • Jane Jourdan
  • Robert G. Gourdie
Article

Abstract

Gap junctions (GJs) are aggregates of channels that provide for direct cytoplasmic connection between cells. Importantly, this connection is thought responsible for cell-to-cell transfer of the cardiac action potential. The GJ channels of ventricular myocytes are composed of connexin43 (Cx43). Interaction of Cx43 with zonula occludens-1 (ZO-1) is localized not only at the GJ plaque, but also to the region surrounding the GJ, the perinexus. Cx43 in the perinexus is not detectable by immunofluorescence, yet localization of Cx43/ZO-1 interaction to this region indicated the presence of Cx43. Therefore, we hypothesized that Cx43 occurs in the perinexus at a lower concentration per unit membrane than in the GJ itself, making it difficult to visualize. To overcome this, the Duolink protein–protein interaction assay was used to detect Cx43. Duolink labeling of cardiomyocytes localized Cx43 to the perinexus. Quantification demonstrated that signal in the perinexus was lower than in the GJ but significantly higher than in nonjunctional regions. Additionally, Duolink of Triton X-100-extracted cultures suggested that perinexal Cx43 is nonjunctional. Importantly, the voltage gated sodium channel Nav1.5, which is responsible for initiation of the action potential, was found to interact with perinexal Cx43 but not with ZO-1. This work provides a detailed characterization of the structure of the perinexus at the GJ edge and indicates that one of its potential functions in the heart may be in facilitating conduction of action potential.

Keywords

Connexin43 Duolink Gap junction Hemichannel Nav1.5 Perinexus Sodium channel 

Notes

Acknowledgments

This work was supported in part by grants from the National Institutes of Heath (RO1 HL56728-10A2 to RGG, RO11DE019355-1 RGG subcontract, F30 HL095320-01 RGG mentor, and 5P20RR016434-07 RGG mentor), and an AHA Grant-in-Aid (RGG).

Supplementary material

232_2012_9465_MOESM1_ESM.tif (188 kb)
Supplementary material 1 (TIFF 188 kb)

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • J. Matthew Rhett
    • 1
  • Emily L. Ongstad
    • 1
  • Jane Jourdan
    • 1
  • Robert G. Gourdie
    • 1
  1. 1.Department of Regenerative MedicineMedical University of South CarolinaCharlestonUSA

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