The fine structure of bovine nasal cartilage. Extraction as a technique to study proteoglycans and collagen in cartilage matrix.
Bovine nasal cartilage was studied by electron microscopy before and after extraction with 4 M guanidinium chloride or 1.9 M CaCl(2). These solvents removed matrix granules, basophilia, and 85% of the proteoglycan complex, measured as hexuronate. Simultaneously, many collagen fibrils were disaggregated into component microfibrils (approximately 40 A thick). In contrast to the above solvents, exhaustive extraction with 0.5 M guanidinium chloride removed 20% of the proteoglycan complex, and matrix granules were reduced in size but not in number. Extraction with 4 M CaCl(2) removed only 10% of the proteoglycan complex, did not remove matrix granules, and caused the normal banding pattern of collagen to disappear. The banding was restored by further treatment with trypsin. Trypsin, before or after 4 M CaCl(2), removed matrix granules and 90% of the proteoglycan complex. We conclude that matrix granules are an electron microscopic representation of the proteoglycan complex, and consist of more than one proteoglycan macromolecule. It would appear that 4 M guanidinium chloride and 1.9 M CaCl(2), in addition to removing most of the proteoglycan complex, also disaggregate some of the collagen fibrils into their component microfibrils.
D000818:Animals; D001491:Basophils; D002122:Calcium Chloride; D002356:Cartilage / Q000166:cytology*; D002417:Cattle; D002452:Cell Count; D002467:Cell Nucleus; D002712:Chlorides; D003094:Collagen / Q000032:analysis*; D003593:Cytoplasm; D004721:Endoplasmic Reticulum; D006003:Glycogen / Q000032:analysis; D006056:Golgi Apparatus; D006651:Histocytochemistry; D006652:Histological Techniques; D046911:Macromolecular Substances; D008722:Methods; D008854:Microscopy, Electron; D008928:Mitochondria; D009300:Nasal Septum / Q000166:cytology*; D011134:Polysaccharides / Q000032:analysis*; D012997:Solvents; D013194:Staining and Labeling; D014357:Trypsin; D014574:Uronic Acids / Q000032:analysis
D002712:Chlorides; D046911:Macromolecular Substances; D011134:Polysaccharides; D012997:Solvents; D014574:Uronic Acids; D006003:Glycogen; D003094:Collagen; D014357:Trypsin; D002122:Calcium Chloride