Krupczak-Hollis Katherine, Wang Xinhe, Kalinichenko Vladimir V, Gusarova Galina A, Wang I-Ching, Dennewitz Margaret B, Yoder Helena M, Kiyokawa Hiroaki, Kaestner Klaus H, Costa Robert H
Department of Biochemistry and Molecular Genetics, College of Medicine, University of Illinois at Chicago, Chicago, IL 60607-7170, USA.
Dev Biol. 2004 Dec 1;276(1):74-88. doi: 10.1016/j.ydbio.2004.08.022.
Conditional deletion of the mouse Forkhead Box (Fox) m1b targeted allele in adult hepatocytes (Foxm1, previously called HFH-11B, Trident, Win, or MPP2) demonstrated that the Foxm1b transcription factor is essential for hepatocyte mitosis during liver regeneration. To determine the role of Foxm1b in liver development, we have generated Foxm1b -/- mice that deleted the Foxm1b exons encoding the winged helix DNA binding and transcriptional activation domains. Here, we show that all of the Foxm1b -/- embryos died in utero by 18.5 days postcoitum (dpc). Embryonic Foxm1b -/- livers displayed a 75% reduction in the number of hepatoblasts, resulting from diminished DNA replication and a failure to enter mitosis causing a polyploid phenotype. Reduced hepatoblast mitosis was associated with decreased protein levels of the Polo-like kinase 1 and Aurora B kinase, which phosphorylate regulatory proteins essential for orchestrating mitosis and cytokinesis. Diminished proliferation of Foxm1b -/- hepatoblasts contributed to abnormal liver development with significant reduction in the number of large hepatic veins compared to embryonic wild-type (WT) liver. Furthermore, embryonic Foxm1b -/- livers did not develop intrahepatic bile ducts, and these presumptive biliary hepatoblasts failed to express either biliary cytokeratins or nuclear levels of hepatocyte nuclear factor 1beta. These results suggest that Foxm1b is critical for hepatoblast precursor cells to differentiate toward biliary epithelial cell lineage. Finally, we used a hepatoblast-specific Cre recombinase transgene to mediate deletion of the Foxm1b fl/fl allele in the developing liver, and these embryos died in utero and exhibited diminished hepatoblast proliferation with similar abnormalities in liver morphogenesis, suggesting that the defect in liver development contributed to embryonic lethality.
在成年肝细胞中条件性删除小鼠叉头框(Fox)m1b靶向等位基因(Foxm1,以前称为HFH-11B、Trident、Win或MPP2)表明,Foxm1b转录因子对肝脏再生过程中的肝细胞有丝分裂至关重要。为了确定Foxm1b在肝脏发育中的作用,我们构建了Foxm1b-/-小鼠,其缺失了编码翼状螺旋DNA结合和转录激活结构域的Foxm1b外显子。在此,我们表明所有Foxm1b-/-胚胎在子宫内于妊娠18.5天(dpc)死亡。胚胎期Foxm1b-/-肝脏中肝母细胞数量减少了75%,这是由于DNA复制减少以及无法进入有丝分裂导致多倍体表型所致。肝母细胞有丝分裂减少与Polo样激酶1和Aurora B激酶的蛋白水平降低有关,这两种激酶可磷酸化协调有丝分裂和胞质分裂所必需的调节蛋白。与胚胎野生型(WT)肝脏相比,Foxm1b-/-肝母细胞增殖减少导致肝脏发育异常,肝大静脉数量显著减少。此外,胚胎期Foxm1b-/-肝脏未发育出肝内胆管,这些假定的胆管肝母细胞未能表达胆管细胞角蛋白或肝细胞核因子1β的核水平。这些结果表明,Foxm1b对于肝母细胞前体细胞向胆管上皮细胞谱系分化至关重要。最后,我们使用肝母细胞特异性Cre重组酶转基因介导发育中的肝脏中Foxm1b fl/fl等位基因的删除,这些胚胎在子宫内死亡,并表现出肝母细胞增殖减少以及肝脏形态发生类似异常,表明肝脏发育缺陷导致胚胎致死。