Evidence for the involvement of calpain in cataractogenesis in Shumiya cataract rat (SCR).
The Shumiya cataract rat (SCR) is a hereditary cataract model in which lens opacity appears spontaneously in the nuclear and perinuclear portions at 11-12 weeks of age. It was found that the proteolysis of some crystallins and cytoskeletal proteins is significantly enhanced in cataractous SCR lenses. The calcium concentrations in cataractous lenses rise markedly with age as compared with control lenses and the autolytic product of calpain is also detected in cataractous lenses. In order to provide direct evidence for the involvement of calpain in the proteolytic modification of lens proteins, we developed antibodies exclusively specific to the proteolytic products of some lens proteins produced by the action of calpain and analyzed their degradation during cataractogenesis in SCR by Western blotting and immunohistochemical staining. The results demonstrate that calpain participates in the proteolytic modification of lens proteins, at least alpha-crystallin (A and B chain), betaB1-crystallin, and alpha-fodrin. The proteolytic products formed by the action of calpain on these proteins are detected in cataractous lenses of SCR as young as 8 weeks of age and accumulate with age. It was also found that betaB1-crystallin, originally a soluble protein, is converted to an insoluble form by limited calpain proteolysis. The chaperon-like activity of alpha-crystallin from control lens is markedly reduced by calpain proteolysis in vitro, and alpha-crystallin in opaque lens that has already undergone proteolysis by calpain shows significantly reduced chaperon-like activity. Immunohistochemical studies reveal that the area where the calpain-mediated alpha-crystallin proteolysis is in progress coincides well with the area developing and destined to develop the opacification. These results strongly suggest that calpain may contribute to lens opacification during cataract formation in SCR.
Inomata
M
M
Department of Enzyme Biochemistry, Tokyo Metropolitan Institute of Gerontology, Japan. minomata@center.tmig.or.jp
Nomura
K
K
Takehana
M
M
Saido
T C
TC
Kawashima
S
S
Shumiya
S
S
D000375:Aging; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000502:physiology*; D002352:Carrier Proteins / Q000378:metabolism; D002386:Cataract / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D003459:Crystallins / Q000378:metabolism; D007908:Lens, Crystalline / Q000378:metabolism*; D008840:Microfilament Proteins / Q000378:metabolism; D018832:Molecular Chaperones / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D013049:Spectrin
D000911:Antibodies, Monoclonal; D002118:Calcium; D002154:Calpain; D002352:Carrier Proteins; D003459:Crystallins; D008840:Microfilament Proteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D013049:Spectrin