Biosynthesis of Sulphated Macromolecules by Rabbit Lens Epithelium. II. Relationship to Basement Membrane Formation

• Published in: The Journal of cell biology 1984-09, Vol.99 (3), p.861-869
• Main Authors: Heathcote, J. Godfrey, Bruns, Romaine R., Orkin, Roslyn W.
• Format: Article
• Language: English
• Subjects: Animals; Ascorbic Acid - pharmacology; Basement membrane; Basement Membrane - metabolism; Basement Membrane - ultrastructure; Biological and medical sciences; Cell Fractionation; Cell lines; Cell metabolism, cell oxidation; Cell physiology; Cells, Cultured; Chondroitin Sulfate Proteoglycans - biosynthesis; Collagens; Cultured cells; Epidermal cells; Epithelial cells; Epithelium - drug effects; Epithelium - metabolism; Female; Fundamental and applied biological sciences. Psychology; Gels; Glycosaminoglycans - biosynthesis; Heparan Sulfate Proteoglycans; Heparitin Sulfate - biosynthesis; Hydrochlorides; Lens, Crystalline - metabolism; Macromolecules; Male; Molecular and cellular biology; Proline - metabolism; Proteoglycans; Proteoglycans - biosynthesis; Rabbits; Sulfates - metabolism; Sulfur Radioisotopes; Tritium
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.99.3.861

Rabbit lens epithelial cells display a similar "cobblestone" morphology and produce the same complement of sulphated macromolecules whether grown on plastic or glass, dried films of gelatin or type IV collagen with laminin, or on gels of type I collagen. There was no evidence of basement membrane formation by these cells when they were grown on plastic, glass, or dried films. In contrast, cultures that had been grown on gels deposited a discrete basement membrane that followed the contours of the basal surfaces of the cells and in addition, they secreted amorphous basement membrane-like material that diffused into the interstices of the gel and associated with the collagen fibrils of the gel. A significant proportion (∼70%) of the heparan sulphate proteoglycan fraction that was secreted into the culture medium (fraction MI) when the cells were grown on plastic became associated with the cell-gel layer in the gel cultures. Further, when basement membrane was isolated by detergent extraction, >90% of the35S-labeled material present was in this heparan sulphate proteoglycan.