Subject(s)
Cathepsins/isolation & purification , Mesentery/enzymology , Neoplasms, Experimental/enzymology , Animals , Cathepsin B , Cytoplasmic Granules/enzymology , Fibroblasts/enzymology , Histiocytes/enzymology , Immunoenzyme Techniques , Microscopy, Phase-Contrast , Neoplasm Transplantation , RabbitsABSTRACT
Localization of cathepsin B was studied in the rabbit mesentery during invasion of V2 carcinoma cells. Cathepsin B was visualized immunohistochemically by using monospecific sheep antibodies and the avidin-biotin-peroxidase complex (ABC) method. Horizontal and vertical semithin Epon embedded sections of stained mesenteries showed that histiocytes always displayed the strongest staining reaction independently of the presence of V2 carcinoma cells. Fibroblasts, mesothelial cells and the invaded V2 cells were less stained. Strongly stained peritoneal monocytes were frequently found on the surface of the mesentery in association with tumor foxi. The role of these various cathepsin B containing cells with respect to extracellular matrix degradation during tumor invasion in the mesentery is not clear; some aspects of this problem are presented in the discussion.
Subject(s)
Cathepsins/analysis , Mesentery/enzymology , Peritoneal Neoplasms/enzymology , Animals , Breast Neoplasms/enzymology , Cathepsin B , Cysteine Endopeptidases , Endopeptidases/metabolism , Histocytochemistry , Humans , Immunoenzyme Techniques , Injections, Intraperitoneal , Neoplasm Invasiveness , Rabbits , Sheep , Staining and LabelingABSTRACT
Visualization of rabbit cathepsin B was achieved utilizing monospecific sheep antibodies and the avidin-biotin-peroxidase complex (ABC) method. This technique was applied to stain 1) paraffin sections of the liver, 2) fixed fibroblasts from tissue culture, and 3) fixed mesenteries. Cathepsin B was found to be localized within cells of the lining of the liver sinusoids (most probably Kupffer cells), in perinuclear granules of cultured fibroblasts, and within histiocytes of the mesentery. The results demonstrate that the method permits precise and highly sensitive localization of cathepsin B within cells and tissues. Compared to fluorescent staining of cathepsin B, the ABC method has the advantage that routine paraffin sections can be stained, and that all the orthodox histological staining procedures can still be carried out.
