Electron microscopic study of distribution of proteoglycans in bovine cornea and sclera

To study differences in the distribution of proteoglycans and their relationships to collagen fibrils in the cornea and sclera, bovine cornea and sclera were examined histochemically using the ruthenium red (RR) staining method. RR-positive granules (30 nm in diameter) were present in association with fine filamentous materials (3 nm in diameter and 30-100 nm in length) in the interfascicular spaces of collagen bundles in both the corneal stroma and sclera. The amount of these materials was smaller in the sclera than in the cornea. The characteristic band-like arrangement of RR-positive granules connected by filamentous materials at intervals of 80-100 nm was found only in the cornea. In enzyme digestion experiments, tissue sections were treated by chondroitinase ABC, AC, and keratanase before RR staining. The RR-positive granules and their associated filamentous materials were darkly stained after chondroitinase AC or keratanase digestion, but displayed markedly lighter staining after chondroitinase ABC digestion. These results indicate that RR-positive granules and filamentous materials contain dermatan sulfate proteoglycan.