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An Autoradiographic Demonstration of the Effect of the Corneal Epithelium on Amino Acid Incorporation into the Insoluble Constituents of the Corneal Stroma

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The Rockefeller University Press
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Keywords
  • Brief Notes
Disciplines
  • Biology

Abstract

0135.tif BRIEF NOTES 135 An Autoradiographic Demonstration of the Effect of the Corneal Epithelium on Amino Acid Incorporation into the Insoluble Constituents of the Corneal Stroma.* BY HEINZ I I~RR~AN~++ AND DAVID S. LOVE.+ + (From the Laboratory of Chemical Embryology, Department of Pediatrics, University of Colorado Medical School, Denver.) § In previous experiments, (1, 2) it was observed that the incorporation of glycine-l-C 14 into protein fractions of the corneal stroma of the chick eye was greatly reduced in the absence of the corneal epithelium. Partial removal of the epithelium from the surface of the stroma led to a proportional re- duction in the incorporation of the label into the stromal proteins. This finding suggested that the effect of the epithelium was restricted to a limited, closely adjacent area. I t seemed desirable, there- fore, to ascertain autoradiographically the extent of the epithelial influence on the uptake of label into the stroma. Such autoradiographs actually do show that the area in the stroma with high label uptake coIcesponds very closely to the area of the epithelial cover. Methods The dissection and the conditions for the uptake of glycine-l-C 14 were described previously (1, 2). The chick corneas were obtained from 8-day hatched chicks of the High Line strain. Each cornea was suspended in a 10 ml. beaker in 2 ml. of a synthetic tissue culture nutrient solution containing glucose, amino acids, vitamins, and electrolytes. Twelve #g. in a solution of 0.01 ml. of glycine-l-C t4 (Nuclear, Chicago) correspond- ing to an activity of 0.6 #C. were added to the samples at the same time, and incubation at 37°C. in a Dubnoff shaker was started immediately after addition of the label. In the present series of experiments, one-half of the epithelium was removed with a sharp scalpel before incubation with the label. After an incubation period of 1 hour, the corneas were briefly washed with Ringer's solution. Unbound

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