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American Journal of Pathology, Vol 115, 31-35, Copyright © 1984 by American Society for Investigative Pathology

REGULAR ARTICLES

Multiparameter flow cytometric analysis of hepatic nuclear RNA and DNA of normal and hepatotoxin-treated mice

PJ Higgins, M Piwnicka, Z Darzynkiewicz and MR Melamed

Combined immunohistologic and flow cytometric methods were used to monitor the regenerative response of the mouse liver following intraperitoneal administration of the hepatonecrotic agent carbon tetrachloride (CCl4). A slight increase in the percentage of liver nuclei engaged in DNA synthesis (S-phase population) was evident 1 day after CCl4. The percentage of S-phase nuclei increased thereafter, lasting until day 6 after CCl4 with a peak at day 2 after CCl4. The method of sampling used (every 24 hours) places the period of maximal DNA synthetic activity between 48 hours and 72 hours after inoculation of the hepatonecrotic agent; at 48 hours the number of S-phase nuclei was approximately five times higher than the nonregenerating liver. Modified multiparametric flow cytofluorimetric analysis of hepatic nuclei indicated that early repopulation of the liver in response to CCl4-induced necrosis involved the selective proliferation of a specific subpopulation of liver cells which contained diploid nuclei of high RNA content. These nuclei were also observed in control animals, suggesting the existence of a subcompartment of 2C DNA content G1 hepatocytes potentially programmed for replacement proliferation.


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Copyright © 1984 by the American Society for Investigative Pathology.