MATURATION-DEPENDENT CHANGES IN THE REGULATION OF LIVER-SPECIFIC GENE-EXPRESSION IN EMBRYONAL VERSUS ADULT PRIMARY LIVER CULTURES

During rat liver development, which starts on day 10 of embryogenesis (E10), and until E15, all parenchymal cells are thought to be a homogeneous population of bipotential progenitors, able to give rise to both hepatocytes and bile duct epithelial cells. We established primary liver cultures from embryonic livers at various developmental stages, from E14 to neonates, as well as adult rats. Gene expression and regulation by three known differentiating agents, heparin, dimethylsulfoxide (DMSO), and sodium butyrate, were examined in these primary cultures. Alpha-fetoprotein (alpha-FP), albumin, gamma-glutamyltranspeptidase (GGT), and glutathione-S-transferase-P (Yp) were expressed by cultured liver cells through fetal development, whereas insulin-like growth factor-II (IGF II) receptor, expressed in fetal parenchymal cells, was not present in cultured neonatal cells. Heparin increased alpha-FP levels in fetal liver cells, but not in cells obtained after birth. The expression of GGT and Yp was coordinately regulated. The two genes were up-regulated by sodium butyrate and down-regulated by DMSO in cultured liver cells from all embryonal ages tested. However, the regulation of these two genes by sodium butyrate and DMSO was not apparent in neonatal and adult liver cultures. Sodium butyrate increased alpha-FP and albumin mRNA expression in E14 and E15 cells, but not in E16, neonatal or adult cultures, and its addition caused heterogenous expression of albumin. We conclude that the regulation of gene expression in primary liver cultures by the three agents tested is altered after birth. In fetal cells, sodium butyrate serves as a strong general differentiation agent, increasing expression of sets of genes characteristic for both hepatocytes and bile duct cells. By contrast, DMSO suppresses sets of genes expressed in bile-duct cells, e.g. GGT, Yp, and cytokeratin 19 (CK19).