Immunohistochemical localization of microcystin-LR in the liver of mice: a study on the pathogenesis of microcystin-LR-induced hepatotoxicity.
The relationship between the intralobular sites of hepatotoxic injury and the distribution of microcystin-LR (MCLR), an inhibitor of protein phosphatases 1 (PP1) and 2A (PP2A), was examined using an immunohistochemical method with a monoclonal antibody specific to MCLR on the livers of mice receiving a single i.p. injection of the MCLR. Immunoblotting and high-performance liquid chromatography analyses of liver extracts were also performed to determine the binding form of MCLR to PP1 and PP2A (MCLR-PP1/PP2A adducts) and free MCLR. Immunohistochemistry revealed a discernible intensity of staining in the centrilobular regions where hemorrhage and apoptosis occurred. In these regions, immunopositivity was evident in the cytoplasm and nuclei of the hepatocytes; some apoptotic cells were also immunopositive. In contrast, coagulative necrosis, which was mainly evident in the midlobular regions, was completely negative. Analysis of liver extracts demonstrated MCLR-PP1/PP2A adducts, but free MCLR was below detection limit. These results suggest that the immunohistochemical localization of MCLR in centrilobular hepatocytes is closely associated with the onset of hemorrhage and apoptosis and is related to adduct formation. The occurrence of coagulative necrosis however might also be related to other factors such as ischemia/hypoxia.
Yoshida
T
T
Research Institute for Biosciences, Faculty of Pharmaceutical Sciences, University of Tokyo, Japan. zanken@iinet.or.jp
Makita
Y
Y
Tsutsumi
T
T
Nagata
S
S
Tashiro
F
F
Yoshida
F
F
Sekijima
M
M
Tamura
S
S
Harada
T
T
Maita
K
K
Ueno
Y
Y
D000818:Animals; D001427:Bacterial Toxins / Q000008:administration & dosage / Q000493:pharmacokinetics; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000493:pharmacokinetics*; D005260:Female; D015151:Immunoblotting; D007150:Immunohistochemistry; D007274:Injections, Intraperitoneal; D008099:Liver / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D008168:Lung / Q000473:pathology; D008387:Marine Toxins; D051379:Mice; D008807:Mice, Inbred BALB C; D052998:Microcystins; D010456:Peptides, Cyclic / Q000008:administration & dosage / Q000493:pharmacokinetics*; D010749:Phosphoprotein Phosphatases / Q000032:analysis; D013997:Time Factors
D001427:Bacterial Toxins; D004791:Enzyme Inhibitors; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D010749:Phosphoprotein Phosphatases; C057862:cyanoginosin LR