In vivo cell lineage analysis in cyproterone acetate-treated rat liver using genetic labeling of hepatocytes

Abstract

Genetic labeling using recombinant retroviruses is a powerful strategy for the study of cell lineage in the liver. However, this type of vector is only able to infect dividing cells. The synthetic steroid cyproterone acetate (CPA) is mitogenic and carcinogenic in the adult rat liver. In this study, we used retroviral vectors carrying the nuclear targeted ␤-galactosidase gene to selectively label and follow the fate of hepatocytes dividing on administration of CPA. Labeled cells as well as those in mitosis were preferentially located around the portal tract, whereas apoptotic bodies were predominant in the pericentral area.

Long term phenobarbital administration does not promote the multiplication of hepatocytes replicating after single cyproterone acetate administration

“…1). This figure is in keeping with our previous study of gene marking after CPA administration (Auvigne et al, 2002). Also, as previously observed, the positive hepatocytes in the day 7 biopsy were either isolated or gathered in small clusters of 2 to 3 cells, preferentially located in periportal and mediolobular areas.…”

“…The h-galactosidase retroviral vector (10 8 infectious viral particles/ ml) was injected twice at 24 and 30 hours after the second administration of CPA since we previously documented that this delay corresponded to the peak of mitoses in the liver after acute CPA administration (Auvigne et al, 2002). Because retroviral vectors are only able to infect, and hence to Fig.…”

“…Recently, we used genetic labeling of hepatocytes to decipher the relationship between the immediate mitogenic and tumor initiating activities of CPA administered acutely. We demonstrated that the initiating and mitogenic activities of CPA were directed towards different hepatocyte populations (Auvigne et al, 2002). In that study, we used onco-retroviral vectors carrying the h-galactosidase marker gene to selectively label the hepatocytes dividing upon the effect of acute CPA administration.…”

Characterization of Multinuclear Hepatocytes Induced in Rats by Mitemcinal (GM-611), an Erythromycin Derivative

“…Several studies indicate that the type of shift is dependent on the cause (Seglen 1997). Although the ploidy was not considered in the present study, the sharp shift toward mononuclear cells was thought to be similar to shifts seen in regenerating rat liver after partial hepatectomy (Auvigne et al 2002;Frederiks et al 1990;Gerlyng et al 1993;Melchiorri et al 1993;Seglen 1997;Wheatley 1972), which is similar to shifts with certain drugs (Gerlyng et al 1994;Seglen 1997).…”

Mature hepatocytes are the source of small hepatocyte-like progenitor cells in the retrorsine model of liver injury

“…Retroviral vectors allow a stable integration and expression of the marker gene and have already been successfully used to study cell lineage in various organs [20,21]. Using this approach we previously studied cell lineage in the liver in physiological and pathological conditions [22][23][24][25]. The use of the b-galactosidase gene from E. coli coupled to a nuclear localisation signal allows a rapid and sensitive histochemical detection of the marker that is concentrated around the nuclear envelope.…”