Fate tracing of mature hepatocytes in mouse liver homeostasis and regeneration
Yann Malato, … , Dirk Grimm, Holger Willenbring
Yann Malato, … , Dirk Grimm, Holger Willenbring
Published November 21, 2011
Citation Information: J Clin Invest. 2011;121(12):4850-4860. https://doi.org/10.1172/JCI59261.
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Fate tracing of mature hepatocytes in mouse liver homeostasis and regeneration

Abstract

Recent evidence has contradicted the prevailing view that homeostasis and regeneration of the adult liver are mediated by self duplication of lineage-restricted hepatocytes and biliary epithelial cells. These new data suggest that liver progenitor cells do not function solely as a backup system in chronic liver injury; rather, they also produce hepatocytes after acute injury and are in fact the main source of new hepatocytes during normal hepatocyte turnover. In addition, other evidence suggests that hepatocytes are capable of lineage conversion, acting as precursors of biliary epithelial cells during biliary injury. To test these concepts, we generated a hepatocyte fate-tracing model based on timed and specific Cre recombinase expression and marker gene activation in all hepatocytes of adult Rosa26 reporter mice with an adenoassociated viral vector. We found that newly formed hepatocytes derived from preexisting hepatocytes in the normal liver and that liver progenitor cells contributed minimally to acute hepatocyte regeneration. Further, we found no evidence that biliary injury induced conversion of hepatocytes into biliary epithelial cells. These results therefore restore the previously prevailing paradigms of liver homeostasis and regeneration. In addition, our new vector system will be a valuable tool for timed, efficient, and specific loop out of floxed sequences in hepatocytes.

Authors

Yann Malato, Syed Naqvi, Nina Schürmann, Raymond Ng, Bruce Wang, Joan Zape, Mark A. Kay, Dirk Grimm, Holger Willenbring

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Figure 1

Rapid marker gene activation in all hepatocytes of adult R26R-EYFP and R26R mice injected with AAV8-Ttr-Cre.

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Rapid marker gene activation in all hepatocytes of adult R26R-EYFP and R...
(A) Results of AAV8-Ttr-Cre vector titration by dot blot and qPCR. The titer was approximately 4 × 1012 viral genomes/ml in both dot blot and qPCR analysis (Supplemental Figure 1, A and B). Data represent mean ± SEM. (B) Dose-finding strategy. Livers were analyzed by immunostaining for EYFP 5 days after injection of different AAV8-Ttr-Cre doses into the tail veins of adult R26R-EYFP mice. (C) 1.5 × 1011 viral genomes were not sufficient to activate EYFP expression (red) in all hepatocytes. 4 × 1011 viral genomes activated EYFP expression in all hepatocytes. Nuclei were stained with DAPI (blue). (D) X-gal staining of livers of male and female R26R mice 48 hours after injection of 4 × 1011 viral genomes shows β-gal expression (blue) in all hepatocytes. 15 liver sections from 3 mice were analyzed for each experiment. Original magnification, ×200.