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(Investigative Ophthalmology and Visual Science. 2003;44:672-679.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.02-0018

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Activation of Metallothioneins and {alpha}-Crystallin/sHSPs in Human Lens Epithelial Cells by Specific Metals and the Metal Content of Aging Clear Human Lenses

John R. Hawse,1 Jonathan R. Cumming,1 Brian Oppermann,1 Nancy L. Sheets,1 Venkat N. Reddy,2 and Marc Kantorow1

1From the Department of Biology, West Virginia University, Morgantown, West Virginia; and the 2Department of Ophthalmology and Visual Sciences, University of Michigan, Ann Arbor, Michigan.

PURPOSE. To identify those metallothionein and {alpha}-crystallin/small heat-shock genes induced by toxic metals in human lens cells and to evaluate the levels of these metals between young and aged human lenses.

METHODS. Human SRA01/04 and primary human lens epithelial cells were cultured and exposed to Cd2+, Cu2+, and Zn2+. The levels of lens metallothioneins (Ig, If, Ih, Ie, and IIa) and {alpha}-crystallin/small heat-shock ({alpha}A-crystallin, {alpha}B-crystallin, and HSP27) genes were analyzed by semiquantitative and quantitative competitive RT-PCR. The content of aluminum, cadmium, calcium, chromium, copper, iron, lead, magnesium, manganese, nickel, potassium, sodium, and zinc in young (mean, 32.8 years), middle-aged (mean, 52.3 years), and old (mean, 70.5 years) human lenses was analyzed by inductively coupled plasma-emission spectroscopy.

RESULTS. Lens metallothioneins (Ig, If, Ih, Ie, and IIa) and {alpha}-crystallin/small heat-shock genes ({alpha}A-crystallin, {alpha}B-crystallin, and HSP27) were differentially induced by specific metals in SRA01/04 human lens epithelial cells. Cd2+ and Zn2+, but not Cu2+, induced the metallothioneins, whereas Cd2+ and Cu2+, but not Zn2+, induced {alpha}B-crystallin and HSP27. {alpha}A-crystallin was induced by Cu2+ only. Similar responses of the metallothionein IIa gene were detected in identically treated primary human lens epithelial cells. Cd2+ and Zn2+ induced metallothionein IIa to five times higher levels than metallothionein Ig. Of 13 different metals, only iron was altered, exhibiting an 81% decrease in old versus young lenses.

CONCLUSIONS. Induction of metallothioneins and {alpha}-crystallin/small heat shock proteins by different metals indicates the presence of metal-specific lens regulatory pathways that are likely to be involved in protection against metal-associated stresses.





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