Transketolase gene expression in the cornea is influenced by environmental factors and developmentally controlled events

Abstract

Purpose: Transketolase (TKT) has been proposed to be a corneal crystallin, and its gene and protein are abundantly expressed in the corneal epithelium of several mammals. A marked up-regulation of TKT gene expression coincides with the time of eyelid opening in the mouse. Here, we examined whether exposure to incident light contributes to the up-regulation of TKT gene expression during cornea maturation.

Methods: Mice were raised in either standard light/dark cycling conditions or total darkness. In some cases, subcutaneous injections of epidermal growth factor (EGF) were given beginning on the day of birth to induce early eyelid opening. RNA was prepared from the corneas of mothers and pups and subjected to Northern blot analyses. In addition, the relative levels of TKT mRNA and/or enzyme activity were examined in the corneas of human, bovine, rat, chicken, and zebrafish.

Results: TKT mRNA levels were 2.1-fold higher in the corneas of 25-day-old mouse pups ( 12 days after eyelid opening) that had been born and raised in light/dark conditions compared to pups born and raised in total darkness. By contrast, the level of TKT mRNA in the mature corneas of adult mice maintained in the dark for 2-8 weeks did not vary greatly from those of mice maintained in light/dark conditions. Interestingly, TKT mRNA levels in the corneas of dark-raised mice, although reduced, did exhibit the increase characteristically observed before and after eyelid opening. In addition, TKT mRNA levels were elevated fivefold in the corneas of 28-day-old mice raised in darkness and injected with EGF compared to uninjected mice also deprived of light. The EGF-injected mice opened their eyes 3 days early, and their corneal epithelium did not grossly differ from that of control mice. TKT mRNA and/or enzyme activity was found to be much higher in the corneas than in other tissues of humans, bovines, and rats but was extremely low in the corneas of chicken and zebrafish.

Conclusion: Our studies suggest that both exposure to incident light and events surrounding the process of eyelid opening play a role in the up-regulation of TKT gene expression observed during corneal maturation in mice. Light appears to play a less important role in the mature cornea in maintaining high levels of TKT gene expression. The low levels of TKT in the cornea of chicken and zebrafish support the notion that TKT acts as a taxon-specific enzyme-crystallin in mammals. The involvement of environmental signals for this putative, mammalian cornea crystallin contrasts with the purely developmental signals involved in the up-regulation of the crystallin genes of the lens.