Onset of fibre differentiation in cultured rat lens epithelium under the influence of neural retina-conditioned medium

Abstract

In the intact rat lens, epithelial cells, which are cuboidal and contain alpha-crystallin, give rise to fibre cells, which are elongated and contain alpha-, beta- and gamma-crystallins. Epithelial explants cultured in medium conditioned by bovine neural retinas (BRCM) showed changes analogous to fibre cell differentiation in vivo. The cells became enlarged and elongated and accumulated beta- and gamma-crystallin as well as alpha-crystallin. Labelling studies with [35S]-methionine showed that sequential changes in the synthesis of all three classes of crystallin occurred during culture in BRCM. After two days, synthesis of alpha-crystallin subunits, particularly alpha A and alpha AINS, increased relative to overall protein synthesis. After four days in culture, synthesis of beta-crystallin subunits, identified as beta B1a, beta B4, beta B5 and possibly beta A2, was detected for the first time and between four and eight days gamma-crystallin synthesis became detectable. The time of onset of gamma-crystallin synthesis seemed to show greater experimental variability than did onset of beta-crystallin synthesis. In explants cultured for 10 days in BRCM approximately 25% of new protein synthesis could be attributed to alpha-, beta- and gamma-crystallins. These events were completely dependent on BRCM, suggesting that neural retina secretes a factor(s) which initiates fibre cell differentiation. This culture system appears to be a suitable one for investigating the control of fibre differentiation in vitro.