Abstract
A new, sensitive micromethod for the determination of carbonic anhydrase has been applied to the rabbit cornea. In this technique the concentration of carbonic anhydrase is related to the rate of exchange of 18O between labeled bicarbonate and water during the catalyzed dehydration of bicarbonate. Excised rabbit corneas and extracts prepared from the separated layers of rabbit corneas were placed in 18O-enriched carbonate-bicarbonate buffer solution at pH 9·4 and 25°C. The 18O content of the bicarbonate at periodic intervals was determined by mass spectrometry. Carbonic anhydrase was found to be present in the rabbit corneal endothelium at an estimated cellular concentration of 10 −6 m, the same order of magnitude as in most secretory tissues. No carbonic anhydrase was detected in the stroma or epithelium.
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