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Cellular distribution of lysosomal hydrolase activities in the regenerating rat liver

Cathepsins B and D, β‐galactosidase, and acid phosphatase activities were found to be decreased in the regenerating rat liver, the reduction being maximal around the peak of hepatocyte mitoses (30 h). To investigate whether these changes could be heterogeneously distributed among hepatic cells, total cell populations from control or two‐thirds hepatectomized rat livers were dissociated by the collagenase perfusion technique and analysed by different procedures. Isopycnic centrifugation in a Metrizamide gradient satisfactorily resolved hepatocytes and non‐parenchymal cells from control animals but was not adequate when applied to 30‐h regenerating liver cells. Colchicine treatment of the hepatectomized animals, resulted in substantial accumulation of phase M‐hepatocytes. Subpopulations considerably enriched in fast‐sedimenting phase M‐cells were obtained by sedimentation at 1 g of the total liver cell population, and subsequently analysed by isopycnic equilibration. Phase M‐hepatocytes were shown to have markedly reduced levels of β‐galactosidase, acid phosphatase, and cathepsin B activities in comparison, not only with control hepatocytes, but also with those parenchymal cells which were not metaphase‐arrested in the same regenerating livers. Therefore, in partially‐hepatectomized rats, hepatocytes progressing up to metaphase in the first mitotic cycle exhibited a selective depletion of lysosomal enzyme activities. The mechanism(s) underlying this change remain(s) presently unknown.