Immunocytochemical localization of lysosomal cysteine and aspartic proteinases, and ubiquitin in rat epidermis.
To analyze the degradation system in epidermal cells during their generation, differentiation, and cell death, immunocytochemical localization of lysosomal cysteine and aspartic proteinases, an endogenous cysteine proteinase inhibitor, cystatin beta, and ubiquitin were examined using rat sole skin. By confocal laser microscopy, granular immunodeposits for lysosomal proteinases were well demonstrated in epidermal cells; immunoreactivity for cathepsins B and C was prominent in the lower spinous and basal layers, while that for cathepsins L and D was intense in the upper spinous and granular layers, although immunoreactivity for cathepsin D was also detected in the lower epidermal layers. Immunoreactivity for cathepsin H was weakly detected only in the spinous layer, where there were some intensely immunopositive cells with processes which were also immunopositive for S-100 alpha, indicating that these cells were Langerhans cells. Diffuse immunoreactivity for cystatin beta was intense in the spinous and granular layers and weak in the basal layer. In addition, we also examined the localization of ubiquitin, which is a signal peptide for cytosolic proteolysis; clear-cut granular immunodeposits for ubiquitin were detected in spinous and granular cells, and some were co-localized with cathepsin B immunoreactivity. In the basal layer, mitotic cells were strongly immunopositive for ubiquitin. These results suggest that cysteine and aspartic proteinases are involved in the lysosomal system of the epidermis, showing different distributions in the epidermal layers depending on the enzymes examined. Moreover, ubiquitin may be associated with the cell cycle-dependent degradation in basal cells while it also participates in the non-lysosomal proteolysis and probably, lysosomal proteolysis in the spinous and granular cells.
Sato
K
K
Department of Dermatology, Iwate Medical University, School of Medicine, Morioka, Japan.
Waguri
S
S
Ohsawa
Y
Y
Nitatori
T
T
Kon
S
S
Kominami
E
E
Watanabe
T
T
Gotow
T
T
Uchiyama
Y
Y
D000818:Animals; D000918:Antibody Specificity; D016282:Aspartic Acid Endopeptidases / Q000032:analysis*; D015153:Blotting, Western; D015891:Cystatins / Q000032:analysis; D003546:Cysteine Endopeptidases / Q000032:analysis*; D004817:Epidermis / Q000737:chemistry / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D008247:Lysosomes / Q000737:chemistry*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D014452:Ubiquitins / Q000032:analysis*
D015891:Cystatins; D014452:Ubiquitins; D003546:Cysteine Endopeptidases; D016282:Aspartic Acid Endopeptidases