Survival of fiber cells and fiber-cell messenger RNA in lens of rats maintained on a 50% galactose diet for 45 days.

Previous work from this laboratory has shown that in cataractous lens from rats fed a 50% galactose diet up to 32 days, synthesis of crystallin mRNAs was reduced while synthesis of mRNAs for the non-crystallin proteins appeared to be maintained at equivalent or higher levels than found in the controls. In this study, we find that the population of mRNA from the 45-day cataractous lens included all of the crystallin mRNAs and substantial amounts of mRNAs for the non-crystallin proteins, in particular those proteins of molecular weights ranging from 45,000 to 92,000, and pIs from 5.5 to 7.0. At the 45-day cataractous stage, the observed mRNA products totaled between 100 and 120, while for the control they numbered between 40 and 50. In addition, hybrid-select measurements with a lambda gt11 MP26 clone established that MP26 mRNA persisted in the fiber cells of 11-, 20-, 32- and 45-day galactosemic lens. The data, therefore, suggest that continuous exposure of the lens to galactose apparently leads to significant fluctuations in mRNA synthesis and survival. Indirect immunofluorescence (with a monospecific polyclonal rabbit anti-MP26 antibody) and light-microscopy studies demonstrated that although the cortical fiber cells were swollen and structurally disoriented, they retained their nuclei, while the enucleated fiber cells were absent. The microscopy data also suggest that differentiation of the epithelial cells to fiber cells continued in the cataractous lens. Fiber-cell migration was backward as compared with the control, and the surviving cells were localized within the cortex and appeared to be surrounded by damaged cells or cell debris. It is suggested that persistent fiber-cell survival, and continued synthesis of various classes of mRNA by the nucleated cells in the cataractous lens could explain why reversal of cataracts results in recovery in both cell morphology and transparency in the non-nuclear portion of the lens.