Investigative Ophthalmology & Visual Science, Vol 29, 1713-1725, Copyright © 1988 by Association for Research in Vision and Ophthalmology

ARTICLES AND REPORTS

Membrane and communication properties of tissue cultured human lens epithelial cells

S Stewart, G Duncan, JM Marcantonio and AR Prescott
School of Biological Sciences, University of East Anglia, Norwich, Norfolk, England.

Explant cultures were established from capsule/epithelium preparations from both normal and cataractous lenses to investigate properties of human lens epithelial cells. The cultured cells were found to have similar membrane potentials to whole human lenses and isolated epithelia, and similar free ionic concentrations of potassium, sodium, and calcium (131 mM, 17 mM and 0.8 microM respectively) to whole human lenses. The free ionic concentrations were measured in both cases using neutral resin-filled electrodes. Cellular communication was investigated using electrical (two internal microelectrodes) and dye injection techniques. The electrical resistance of a confluent cell monolayer was approximately 4 M omega when the voltage measuring and current passing microelectrodes were in neighbouring cells or several cell diameters apart. Additionally, Lucifer Yellow dye injected into one cell spread rapidly over a wide area of cells. The cells thus appear to be extremely well coupled. Electrical communication could be disrupted by internal acidification (following exposure to 100% CO2), exposure to 1 mM octanol and by membrane depolarization. In the latter case the blockade was only partial. All uncoupling methods proved to be reversible. The diffusion of Lucifer Yellow dye was also inhibited by internal acidification and exposure to octanol. The sensitivity of both dye and electrical coupling to internal acidification and exposure to octanol is similar to that observed in hepatocytes and other tissues, whereas the effect on cell communication induced by changing the resting potential appears to occur only in a few cell types such as those of embryonic origin.

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