Electron microscopic demonstration of proteoglycans in guinea pig epiphyseal cartilage

Guinea pig epiphyseal cartilage was studied ultrastructurally after staining with ruthenium red or Alcian Blue. Extracellularly, the matrix granules were positively stained with both dyes, whereas no apparent staining of the collagen fibrils occurred. Intracellularly, a positive ruthenium red staining of the Golgi vacuole granules was observed. No Alcian Blue staining of the content of the vacuoles was detected. The fresh, untreated cartilage contained about 1.3% hexosamine on a dry weight basis. About three-fourths thereof represented chondroitin sulfate, the remaining part keratan sulfate and glycoproteins. After digestion of thin cartilage slices with hyaluronidase or chondroitinase ABC the chondroitin sulfate content was less than 50% of that in the buffer-incubated controls. Concomitantly, the matrix granules were reduced in size and number or completely absent. Papain digestion removed more than 90% of the chondroitin sulfate and all matrix granules. From these findings it was concluded that the matrix granules contain proteoglycans. Moreover, it seemed reasonable to assume that the Golgi vacuole granules, at least in part, represent intracellular proteoglycans. Signs of secretion of these granules by exocytosis were occasionally observed.