Lineage tracing of Ascl1-expressing cells in the maternal liver during pregnancy

Abstract

To cope with the high metabolic demands of the body during pregnancy, the maternal liver adapts by increasing its mass and size. This increase is proportional to the increase in total body weight during the course of gestation. The pregnancy-induced maternal liver growth is a result of both hepatocyte hypertrophy and hyperplasia. Microarray analysis of pregnant maternal livers shows markedly different gene expression profiles when compared to a non-pregnant state. Most interesting was the 2,500-fold up-regulation in the mRNA expression of Ascl1, a transcription factor responsible for the differentiation of neural progenitor cells into various neuronal types, during the second half of pregnancy. Our investigation aimed at (1) characterizing the identity of maternal hepatic Ascl1-expressing cells and (2) tracing the fate of Ascl1-expressing cells in the maternal liver during pregnancy. Timed pregnancies were generated and non-pregnant (NP) and pregnant maternal livers were harvested and analysed. To identify the maternal hepatic Ascl1-expressing cells we used the Ascl1GFP/+ reporter mouse line. NP and gestation day 15 (D15) maternal livers were immunostained for green fluorescent protein (GFP). The result shows that GFP-positive, Ascl1-expressing cells are hepatocyte-like cells, which are present in D15 maternal livers, but absent in NP livers. The Rosa26floxstopLacZ/ floxstopLacZ;Ascl1CreERT2/+ mouse line was used to trace the fate of Ascl1-expressing cells during pregnancy. LacZ staining of gestation day 13 (D13) and 18 (D18) maternal livers demonstrates that D13 hepatic Ascl1-expressing cells (labeled with LacZ) undergo hyperplasia to repopulate a large portion of D18 maternal livers. Furthermore, LacZ and HNF4α co-staining of D13 and D18 maternal livers shows the presence of two populations of LacZ-expressing cells: HNF4α+ population and HNF4α- population. HNF4α+ LacZ-expressing cells represent hepatocyte lineage cells that are derived from Ascl1-expressing cells. We observe that, towards the end of pregnancy, a considerable portion of the maternal liver is comprised of hepatocytes derived from Ascl1-expressing cells. Taken together, our preliminary study suggests that pregnancy induces maternal liver turnover via Ascl1-expressing cells.