Immunohistochemical detection of proliferating lipocytes in regenerating rat liver.
For the detection of proliferating lipocytes in regenerating liver, partially hepatectomized rats were injected intraperitoneally with 5-bromo-2'-deoxyuridine (BrdUrd, 50 mg/kg body weight) and killed 1 h later. Acetone-fixed frozen liver sections were used for the simultaneous detection of cytoplasmic desmin and BrdUrd-labelled nuclei of lipocytes using double immunohistochemical procedures. The best results were obtained with the sequences: rabbit anti-desmin----biotinylated anti-rabbit IgG----avidin-biotin-peroxidase complex----3,3'-diaminobenzidine tetrahydrochloride, followed by DNA denaturation----mouse anti-BrdUrd----anti-mouse IgG----peroxidase-anti-peroxidase complex----4-chloro-1-naphthol. With this method, cytoplasmic desmin was stained a brown colour, which sharply contrasted with the blue-stained BrdUrd-labelled nuclei. Unlabelled nuclei appeared green after counterstaining with methyl green. No cross-reaction between immunoreagents of desmin and BrdUrd stainings was observed. The labelling index of lipocytes peaked (25.7 per cent) 48 h after partial hepatectomy, whereas it was 3.7 per cent in normal rat liver.
Tanaka
Y
Y
Alcohol Research Center and Liver Disease and Nutrition Section, Bronx Veterans Administration Medical Center, New York 10468.
Mak
K M
KM
Lieber
C S
CS
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002455:Cell Division / Q000502:physiology; D003593:Cytoplasm / Q000032:analysis; D003893:Desmin / Q000032:analysis; D006498:Hepatectomy; D007150:Immunohistochemistry / Q000379:methods*; D008099:Liver / Q000032:analysis / Q000166:cytology*; D008115:Liver Regeneration / Q000276:immunology; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D003893:Desmin; D001973:Bromodeoxyuridine