Calcium-induced opacification and proteolysis in the intact rat lens.
When intact rat lenses were incubated in artificial aqueous humor in the presence of 1 mM calcium and a sulfhydryl reagent p-chloromercuriphenyl sulfonate (pCMPS) a visible annular opacity developed within 4 hours. Combined photographic and ion-sensitive microelectrode investigations of the lenses demonstrated that the subsequent linear increase in opacification was accompanied by an increase in internal free calcium. Opacities were not observed in lenses incubated in the absence of either pCMPS or calcium. Gel electrophoresis of the soluble and urea-soluble fractions from lenses exposed to 1 mM calcium for periods of up to 14 hours showed no evidence for crystallin degradation and only minor proteolysis of cytoskeletal proteins. When lenses were incubated under identical conditions, but with 5 mM calcium, the degree of opacification increased up to approximately 8 hours and then remained constant. A progressive loss in cytoskeletal proteins was observed which correlated with a further increase in free calcium such that by 14 hours of incubation, when the internal calcium approached 1 mM, most of the spectrin and vimentin present in the cortex of the lens had disappeared. An unidentified 110-kilodalton protein also disappeared from lenses incubated in 5 mM calcium. These results indicate that proteolysis by calcium-dependent enzymes such as calpain may play a significant role in cytoskeletal regulation and metabolism in the lens. A role for cytoskeleton/membrane/crystallin interaction in calcium-induced opacification is discussed.
Truscott
R J
RJ
Australian Cataract Research Foundation, University of Wollongong, New South Wales.
Marcantonio
J M
JM
Tomlinson
J
J
Duncan
G
G
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000502:physiology*; D002154:Calpain / Q000378:metabolism; D002386:Cataract / Q000209:etiology*; D003459:Crystallins / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D007908:Lens, Crystalline / Q000187:drug effects / Q000201:enzymology*; D008564:Membrane Potentials / Q000187:drug effects; D010447:Peptide Hydrolases / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D003459:Crystallins; D003598:Cytoskeletal Proteins; D002731:4-Chloromercuribenzenesulfonate; D010447:Peptide Hydrolases; D002154:Calpain; D002118:Calcium