Immunocytochemical localization of angiotensinogen and cathepsins B, H, and L in rat hepatocytes, with special reference to degradation of angiotensinogen in lysosomes after colchicine.
We examined the effects of bilateral nephrectomy and colchicine treatment on localization and content of angiotensinogen and cathepsins B, H, and L in rat liver using immunohistochemistry, radioimmunoassay, and enzyme assay. Angiotensinogen content increased in the liver of colchicine-treated rats, whereas a clear-cut increase was not detected in the liver of nephrectomized rats. This tendency was consistent with the immunocytochemical results; only perivenous hepatocytes in control and nephrectomized rats were diffusely immunostained by anti-angiotensinogen, whereas perivenous and periportal hepatocytes of colchicine-treated rats were strongly immunostained. Enzyme assay revealed no significant change in activities of cathepsins B, H, and L in liver extracts under these experimental conditions. Immunocytochemical localization of these cysteine proteinases in hepatocytes after colchicine treatment was more widespread in the cytoplasm than that in the control hepatocytes. By electron microscopy, angiotensinogen was localized in smaller vesicles and some larger vesicles (lysosomes) of hepatocytes after colchicine treatment. Double immunostaining demonstrated co-localization of cathepsins B, H, and L with angiotensinogen in lysosomes. These results suggest that cathepsins B, H, and L play a role in the degradation of excess angiotensinogen in hepatocytes of rats after colchicine treatment.
Watanabe
T
T
Department of Anatomy, University of Tsukuba, Japan.
Waguri
S
S
Watanabe
M
M
Ishii
Y
Y
Kominami
E
E
Uchiyama
Y
Y
D000808:Angiotensinogen / Q000378:metabolism*; D000818:Animals; D000918:Antibody Specificity; D002401:Cathepsin B / Q000378:metabolism*; D056655:Cathepsin H; D056668:Cathepsin L; D002403:Cathepsins / Q000378:metabolism*; D003078:Colchicine / Q000378:metabolism*; D003546:Cysteine Endopeptidases; D010450:Endopeptidases; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*; D009392:Nephrectomy; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains
D000808:Angiotensinogen; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D056668:Cathepsin L; C534280:Ctsl protein, rat; D056655:Cathepsin H; C534128:Ctsh protein, rat; D003078:Colchicine