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Age-related changes in fibre differentiation of rat lens epithelial cells in vitro
1Department of Histology and Embryology, University of Sydney, New South Wales, Australia.
Abstract:
Epithelial cells in explants from rat lenses differentiate into fibre cells when cultured with retina conditioned medium (RCM). This differentiation is characterised by cell elongation, the accumulation of alpha-crystallin and the onset of synthesis of the fibre specific beta- and gamma-crystallins. To determine the effect of donor age on the synthesis of beta- and gamma-crystallins in response to culture with RCM, we explanted lens epithelia from 3-, 10- and 21-day-old rats. Explants were trimmed to obtain a population of central epithelial cells and at the end of the culture period crystallin content was measured by ELISA methods and expressed per unit DNA. Explants from 3- and 10-day-old rats showed significant increases in the levels of beta-crystallin/DNA within four days after the addition of RCM. The synthesis of beta-crystallin in explants from 21-day-old rats was delayed until after day 4 of culture, but rose quickly to reach the same levels at 10 days as the explants from younger lenses. The ability of lens epithelia to synthesise gamma-crystallin in response to RCM showed a dramatic decline as the age of the donor rat increased. After 10 days of culture, explants from 10-day-old rats contained only 12% of the amount of gamma-crystallin accumulated by explants from 3-day-old rats. Explants from 21-day-old rats accumulated no significant gamma-crystallin until day 10 of culture, and at this time the amount was significantly less than that accumulated by explants from younger rats.(ABSTRACT TRUNCATED AT 250 WORDS)