August 1970
Volume 9, Issue 8
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Articles  |   August 1970
The Effect of Diamide on Lens Glutathione and Lens Membrane Function
Author Affiliations
  • DAVID L. EPSTEIN
    Howe Laboratory of Ophthalmology, Harvard Medical School, and the Massachusetts Eye and Ear Infirmary, Boston, Mass.
  • JIN H. KINOSHITA
    Howe Laboratory of Ophthalmology, Harvard Medical School, and the Massachusetts Eye and Ear Infirmary, Boston, Mass.
Investigative Ophthalmology & Visual Science August 1970, Vol.9, 629-638. doi:
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      DAVID L. EPSTEIN, JIN H. KINOSHITA; The Effect of Diamide on Lens Glutathione and Lens Membrane Function. Invest. Ophthalmol. Vis. Sci. 1970;9(8):629-638.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

The physiological role of glutathione (GSH), present in exceptionally high concentration in the lens, has never been clearly delineated. Using 86Rb we have studied the cation transport and permeability of rat lenses incubated in vitro with diamide, a relatively specific intracellular GSH oxidant. Lenses incubated without glucose with 0.4 mM. diamide manifest a progressive decrease in GSH with time. In the presence of glucose this effect is minimized, presumably because the GSH level is maintained by the NADPH generated by the pentose shunt mechanism. When lenses are preincubated with diamide without glucose for 2 hours, a 55 to 57 per cent decrease in 86Rb uptake is observed. In addition, diamide also effects a 53 per cent increase in the 86Rb run-out of lenses incubated without glucose. In the presence of glucose both of these effects are minimized. Incubation with dithioihreitol leads to a small but definite increase in the 86Rb uptake of diamide-treated lenses. The observed membrane changes suggest the presence of certain sulfhydryl groups on the lens membrane which are unusally susceptible to oxidation when the GSH level is precipitously reduced. These membrane --SH groups are involved in cation pumping (86Rb uptake) as ivell as in determining the degree of membrane permeability (86Rb run-out). GSH would be necessary to maintain these --SH groups in the reduced form for proper membrane function.

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