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

Mitchell A. Watsky; James L. Rae

doi:10.1007/BF00384726

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

Mitchell A. Watsky, James L. Rae

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	57-63
Number of pages	7
Journal	Cell and Tissue Research
Volume	269
Issue number	1
DOIs	https://doi.org/10.1007/BF00384726
State	Published - Jul 1992
Externally published	Yes

Keywords

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

ASJC Scopus subject areas

Pathology and Forensic Medicine
Histology
Cell Biology

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10.1007/BF00384726

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title = "Dye coupling in the corneal endothelium: effects of ouabain and extracellular calcium removal",

abstract = "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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AU - Rae, James L.

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

Abstract

Keywords

ASJC Scopus subject areas

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