Fortier M, Celton-Morizur S, Desdouets C
Inserm, U1016, Paris, France; CNRS, UMR 8104, Paris, France; Paris Descartes University, Paris, France.
Methods Cell Biol. 2017;137:119-142. doi: 10.1016/bs.mcb.2016.04.006. Epub 2016 May 6.
Polyploidy, the state of having greater than a diploid DNA content (tetraploid, octoploid, etc.) is a characteristic feature of mammalian hepatocytes and accompanies late fetal development and postnatal maturation of the liver. During the weaning period, diploid hepatocytes can engage either into normal cell division cycle giving rise to two diploid hepatocytes or follow a scheduled division program characterized by incomplete cytokinesis. In that case, diploid hepatocytes undergo mitosis, but do not form a contractile ring. Indeed, cleavage-plane specification is never established, because of the deficiencies of actin cytoskeleton reorganization. Furthermore, microtubules fail both to contact the cortex and to deliver their molecular signal, preventing localization and activation of RhoA. Therefore, cytokinesis aborts and a binucleate tetraploid liver cell is generated, which subsequently plays a pivotal role in liver progressive polyploidization. In this chapter, we describe detailed protocols to monitor hepatocyte proliferation and cytokinesis process by in situ and dynamic ex vivo approaches.
多倍体是指具有高于二倍体DNA含量的状态(四倍体、八倍体等),是哺乳动物肝细胞的一个特征,伴随着肝脏胎儿后期发育和出生后成熟过程。在断奶期,二倍体肝细胞可以进入正常细胞分裂周期,产生两个二倍体肝细胞,或者遵循以不完全胞质分裂为特征的预定分裂程序。在这种情况下,二倍体肝细胞进行有丝分裂,但不形成收缩环。事实上,由于肌动蛋白细胞骨架重组的缺陷,分裂平面的确定从未建立。此外,微管既无法接触皮质,也无法传递其分子信号,从而阻止了RhoA的定位和激活。因此,胞质分裂中止,产生一个双核四倍体肝细胞,其随后在肝脏渐进性多倍体化中起关键作用。在本章中,我们描述了通过原位和动态离体方法监测肝细胞增殖和胞质分裂过程的详细方案。