Abstract
To investigate metabolic differences between the central and peripheral cornea the latter including the limbal area, corneas were dissected and examined using phosphorus-31 (31P) nuclear magnetic resonance spectroscopy. Since most31P signals originate from the epithelium,31P spectra of the cornea primarily represent the metabolic state of the epithelium. The spectra of the peripheral cornea showed all phosphorus resonances detected in the whole cornea; in contrast, the central cornea showed no phosphocreatine and glycerophosrylethanolamine, and only low levels of ATP. These results indicate that there is a higher metabolic activity in the peripheral epithelium, especially in the limbal area, than in the central epithelium. To evaluate the metabolic state of corneal epithelium during regeneration, we also examined corneas reepithelializing after 7 mm of central epithelial tissue had been removed by mechanical scraping. Rabbits were killed 24 and 48 h after scraping. The reepithelializing corneas clearly showed an increase in ATP, phosphocreatine, and sugar phosphates with time, although phosphorylcholine remained depressed. These findings suggest that the reepithelializing cornea has an elevated level of energy production and that it may have reached a higher steady state, thereby indicating accelerated metabolism of the epithelium during regeneration.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Brooks WM, Willis RJ (1983)31P nuclear magnetic resonance study of the recovery characteristics of high energy phosphate compounds and intracellular pH after global ischemia in the perfused guinea-pig heart. J Mol Cell Cardiol 15:495–502
Burns RP, Roberts H (1969) Effect of wounding on the corneal epithelial glycogen and related enzyme. Invest Ophthalmol 6:541–547
Ebato B, Friend J, Thoft RA (1987) Comparison of central and peripheral corneal epithelium in culture. Invest Ophthalmol Vis Sci 28:1450–1463
Flaherty JT, Weisfeldt ML, Bulkley BH, Gardner TJ, Gott VL, Jacobs WE (1982) Mechanisms of ischemic myocardial cell damage assessed by phosphorus-31 nuclear magnetic resonance. Circulation 65:561–570
Greiner JV, Kopp SJ, Gillette TE, Glonek T (1983) Phosphatic metabolites of the intact cornea by phosphorus-31 nuclear resonance. Invest Ophthalmol Vis Sci 24:535–542
Greiner JV, Kopp SJ, Glonek T (1984) Nondestructive metabolic analysis of cornea using phosphorus nuclear magnetic resonance. Arch Ophthalmol 102:770–771
Greiner JV, Lass JH, Glonek T (1984) Ex vivo metabolic analysis of eye bank corneas using phosphorus nuclear magnetic resonance. Arch Ophthalmol 102:1171–1173
Greiner JV, Braude LS, Glonek T (1985) Distribution of phosphatic metabolites in the porcine cornea using phosphorus-31 nuclear magnetic resonance. Exp Eye Res 40:335–342
Hayashi K, Kenyon KR (1988) Increased cytochrome oxidase activity in alkali-burned corneas. Curr Eye Res 7:131–138
Henninghausen U, Schmidt-Martens FW, Reim M (1972) Metabolitspiegel und Enzymaktivitaten des Energie liefernden Stoffwechsels im regenerierenden Corneaepithel. Ber Dtsch Ophthalmol Ges 71:95–99
Herrmann H, Hickman FH (1948) Exploratory studies on corneal metabolism. Bull Johns Hopkins Hosp 82:225–250
Kinoshita JH (1962) Some aspects of the carbohydrate metabolism of the cornea. Invest Ophthalmol 1:178–186
Kuwabara T, Perkins DG, Cogan DG (1976) Sliding of the epithelium in experimental corneal wounds. Invest Ophthalmol 15:4–14
Kwok LS (1986) Kinetics of epithelial wound healing in the rabbit cornea. J Infer Deduc Biol 1:1–15
Langham ME (1954) Glycolysis in the cornea of the rabbit. J Physiol 126:396–403
Lemp MA (1976) Cornea and sclera. Arch Ophthalmol 94:473–490
Matsuda H, Smelser GK (1973) Electron microscopy of corneal wound healing. Exp Eye Res 16:427–442
Reim M, Schmidt-Martens FW (1967) Biochemische Veränderungen bei der Vereisung der Hornhaut in vivo. Klin Monatsbl Augenheilkd 150:96–103
Reim M, Conze A, Kaszuba HJ (1982) Adenosine phosphate and glutathione levels in the regenerated corneal epithelium after abrasion and mild alkali burns. Graefe’s Arch Clin Exp Ophthalmol 218:42–45
Schermer A, Galvin S, Sun T-T (1986) Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggest limbal location of corneal stem cells. J Cell Biol 103:49–62
Steuhl K-P, Thiel TJ (1987) Histochemical and morphological study of the regenerating corneal epithelium after limbus-to-limbus denudation. Graefe’s Arch Clin Exp Ophthalmol 225:53–58
Thoft RA, Friend J (1972) Corneal epithelial glucose utilization. Arch Ophthalmol 88:58–62
Thoft RA, Friend J (1975) Biochemical aspects of contact lens wear. Am J Ophthalmol 80:139–145
Thoft RA, Friend J (1976) Corneal epithelial changes during midterm storage. Invest Ophthalmol 15:82–88
Thoft RA, Friend J (1983) The X, Y, Z hypothesis of corneal epithelial maintenance. Invest Ophthalmol Vis Sci 24:1442–1443
Author information
Authors and Affiliations
Additional information
Supported in part by NIH grants EY07620 (HMC) and EY05799 (KRK), and a Grant-in-Aid from Bausch and Lomb (KH)
Rights and permissions
About this article
Cite this article
Hayashi, K., Cheng, HM., Iwasaki, M. et al. Metabolic analysis of reepithelializing rabbit cornea using phosphorus-31 nuclear magnetic resonance spectroscopy. Graefe’s Arch Clin Exp Ophthalmol 228, 73–77 (1990). https://doi.org/10.1007/BF02764295
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02764295