A fluid-phase endocytotic capacity and intracellular degradation of a foreign protein (horseradish peroxidase) by lysosomal cysteine proteinases in the rat junctional epithelium.
We investigated the co-localization of lysosomal cathepsins B, H and L, and horseradish peroxidase (HRP) in junctional epithelial (JE) cells both as a fluid-phase endocytotic marker to demonstrate the fluid-phase endocytotic capacity of JE cells, and to understand the morphological relationships of the endocytosed foreign substances to lysosomal cathepsins in these cells. The diaminobenzidine (DAB) histochemical and cytochemical methods and immunohistochemical avidin-biotin-peroxidase complex and immunocytochemical post-embedding colloidal gold methods were used. Under light microscopy, DAB reaction products based on HRP were found in JE but were rare or absent in the oral sulcular epithelium and oral epithelium. Immunolabeling for cathepsins B and H was found in the granular structures of the cells, but no cathepsin L was identified. With electron microscopy, DAB reaction products, which indicated both HRP and the azurophil granules of neutrophils, were endocytosed into JE cells. Using a post-embedding technique, gold particles indicating HRP were present on the plasma membrane of JE cells, at the periphery of electronlucent vacuoles, and in the electrondense granules. Gold particles indicating cathepsin B or H were found in the electrondense granules. With different sizes of colloidal golds, the co-localization of cathepsin B or H with HRP was indicated only in the electrondense portion of the larger vacuoles consisting of electronlucent and -dense parts. This study provided the first morphological data which indicate that JE has a fluid phase endocytotic capacity, and which suggest that the lysosomal cathepsins B and H are involved in the intracellular degradation of foreign substances invading through the gingival sulcus in JE cells.
Yamaza
T
T
Department of Oral Anatomy I, Faculty of Dentistry, Kyushu University, Japan.
Kido
M A
MA
Kiyoshima
T
T
Nishimura
Y
Y
Himeno
M
M
Tanaka
T
T
D015100:3,3'-Diaminobenzidine; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002401:Cathepsin B / Q000378:metabolism; D056655:Cathepsin H; D056668:Cathepsin L; D002403:Cathepsins / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology / Q000648:ultrastructure; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000201:enzymology / Q000648:ultrastructure; D004705:Endocytosis / Q000502:physiology*; D010450:Endopeptidases; D004792:Enzyme Precursors / Q000378:metabolism; D004846:Epithelial Attachment / Q000201:enzymology* / Q000648:ultrastructure; D004848:Epithelium / Q000201:enzymology / Q000648:ultrastructure; D005260:Female; D005881:Gingiva / Q000166:cytology / Q000201:enzymology; D006651:Histocytochemistry; D006735:Horseradish Peroxidase / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D008247:Lysosomes / Q000201:enzymology*; D008297:Male; D008854:Microscopy, Electron; D051381:Rats; D017208:Rats, Wistar; D014617:Vacuoles / Q000201:enzymology / Q000648:ultrastructure
D015415:Biomarkers; D004792:Enzyme Precursors; D015100:3,3'-Diaminobenzidine; D006735:Horseradish Peroxidase; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D056668:Cathepsin L; C534280:Ctsl protein, rat; D056655:Cathepsin H; C534128:Ctsh protein, rat