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Dye coupling in the corneal endothelium: effects of ouabain and extracellular calcium removal

Cell and Tissue Research volume 269, pages 57–63 (1992)Cite this article

Summary

The effects of ouabain and extracellular calcium removal on gap junctional coupling of isolated rabbit corneal endothelium was examined using a modified dye-spread technique. This technique is a modification of a microelectrode procedure that now utilizes patch electrodes connected to a current-clamp circuit for dye iontophoresis and a shuttering system in the excitation light path to reduce phototoxic effects in the monolayer. It was found that a significant degree of junctional uncoupling occurred after 45 min of exposure to ouabain, quantified as a reduction in the effective diffusion coefficient of injected Lucifer yellow CH: 1.74×10-7 cm2/s (control) versus 0.43×10-7 cm2/s (ouabain-treated). It was also determined that no gap junctional uncoupling occurs after extended exposure (up to 3.5 h) to a calcium-free extracellular environment.

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Affiliations

  1. Department of Physiology and Biophysics, Mayo Foundation, 55905, Rochester, MN, USA

    Mitchell A. Watsky & James L. Rae

  2. Department of Ophthalmology, Mayo Foundation, 55905, Rochester, MN, USA

    James L. Rae

Authors
  1. Mitchell A. Watsky
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  2. James L. Rae
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Watsky, M.A., Rae, J.L. Dye coupling in the corneal endothelium: effects of ouabain and extracellular calcium removal. Cell Tissue Res 269, 57–63 (1992). https://doi.org/10.1007/BF00384726

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  • DOI: https://doi.org/10.1007/BF00384726

Key words

  • Cornea
  • Endothelium
  • Gap junction
  • Ouabain
  • Calcium-free solution
  • Phototoxicity
  • Lucifer yellow
  • Rabbit