Copper-metallothionein distribution in the liver of Long-Evans cinnamon rats: studies on immunohistochemical staining, metal determination, gel filtration and electron spin resonance spectroscopy.
Distribution of metallothionein (MT) and copper ion (Cu) in the liver of LEC (Long-Evans Cinnamon) rats was investigated to examine the relationship between Cu-MT induction and the development of hepatitis followed by hepatocellular carcinomas. Immunohistochemical studies on MT in the liver of LEC rats indicated that MT is accumulated in nuclei and cytosols. Both MT and Cu, estimated by radioimmunoassay and flameless atomic absorption spectrometry, respectively, in subcellular fractions of the liver were found to be concentrated highest in cytosols, followed by nuclei, mitochondria and microsomal fractions. Gel-filtration (Sephadex G-75) studies demonstrated that MT is induced as the Cu-MT form. Furthermore, the Cu-MT fragment purified by the gel-filtration contains the Cu(I)-MT form, as demonstrated by ESR (electron spin resonance) measurements at 77K. These results will be important for understanding the development of hepatitis in LEC rats.
Department of Analytical Chemistry, Kyoto Pharmaceutical University, Japan.
H
H
Sakurai
K
K
Nakajima
H
H
Kamada
H
H
Satoh
N
N
Otaki
M
M
Kimura
K
K
Kawano
T
T
Hagino
D000818:Animals; D002850:Chromatography, Gel; D003300:Copper / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism*; D008297:Male; D008345:Manganese / Q000378:metabolism; D008668:Metallothionein / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D015032:Zinc / Q000378:metabolism
C007121:copper thionein; D008345:Manganese; D003300:Copper; D008668:Metallothionein; D015032:Zinc