The use of colloidal gold complexes in an ultrastructural study of lectin-binding sites and matrix deposition of normal human primary breast epithelium on collagen gels.
Colloidal gold probes were used to study the distribution of peanut agglutinin binding sites and the deposition of extracellular fibronectin and type IV collagen in cultured human breast cells grown on type I collagen gels. Qualitative analysis was performed at the ultrastructural level and appraised in relation to the possible role of peanut agglutinin, fibronectin and type IV collagen as functional markers for distinguishing cell types using this methodology. Peanut agglutinin bound to the surface of cuboidal epithelial cells but not on basal, putative myoepithelial cells in the cell islands, suggesting that it may be a useful functional marker. The binding on the epithelial cells was markedly increased by pre-treatment of the cells with neuraminidase. No correlation was seen between the amount of binding and either the surface topography or cellular ultrastructure. Fibronectin and type IV collagen were demonstrated on the fibrillar network left on the collagen gels after removal of the cell sheet. Any cells still adhering to the gel surface showed no evidence of gold probe binding on their upper surfaces. Examination of the under surfaces of the cell sheet showed gold probe binding equivalent to that found on the gels under the cells. However, it was not proven conclusively which cells produce the fibronectin and type IV collagen.
D001940:Breast / Q000648:ultrastructure*; D002478:Cells, Cultured; D003094:Collagen; D003102:Colloids; D004848:Epithelium / Q000648:ultrastructure; D005260:Female; D005353:Fibronectins / Q000032:analysis; D005782:Gels; D006046:Gold; D006651:Histocytochemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D011975:Receptors, Mitogen / Q000378:metabolism*
D003102:Colloids; D005353:Fibronectins; D005782:Gels; D011975:Receptors, Mitogen; D006046:Gold; D003094:Collagen