Depreter Marianne, Walker Tracy, De Smet Karen, Beken Sonja, Kerckaert Ingrid, Rogiers Vera, Roels Frank
Department of Human Anatomy, Embryology, Histology, and Medical Physics, Ghent University, Godshuizenlaan 4, B-9000 Gent, Belgium.
Histochem J. 2002 Mar-Apr;34(3-4):139-51. doi: 10.1023/a:1020990414190.
In search of factors that regulate the phenotype of the peroxisomal compartment in wild-type liver parenchymal cells, we compared hepatocyte polarity to peroxisome differentiation, using adult liver as the standard. Differentiation parameters were evaluated in a three-dimensional culture model (spheroid), in 'sandwich' and monolayer primary hepatocyte cultures, and in 15.5 and 18.5-day-old foetal rat liver. Peroxisomes, studied by immunohistochemistry, enzyme histochemistry, and catalase specific activity, were better differentiated depending on foetal age (day 18.5 > day 15.5) and culture type (spheroid > sandwich > monolayer). The hepatocyte polarity markers ATP-, ADP-, and AMP-hydrolysing activities were, in all models, mislocalized at the lateral plasma membrane, whereas in contrast the multidrug resistance-associated protein 2 (mrp2) antigen was always correctly immunolocalized at the apical membrane domain. In cultures, the correct secretion of fluorescein (mrp2-mediated) into bile canaliculi was observed. Bile canaliculi (branching, ultrastructure and immunolocalization of the tight-junction associated protein ZO-1), were better differentiated in 18.5 than in 15.5-day-old foetal liver and in spheroid > sandwich > monolayer cultures. Our results show a parallelism between changes of the peroxisomal compartment and bile canalicular structure together with mrp2-mediated secretory function. Distinct polarization characteristics do not necessarily change simultaneously, suggesting different regulatory mechanisms.
为了寻找调节野生型肝实质细胞中过氧化物酶体区室表型的因素,我们以成年肝脏为标准,比较了肝细胞极性与过氧化物酶体分化情况。在三维培养模型(球体)、“三明治”和单层原代肝细胞培养物以及15.5和18.5日龄胎鼠肝脏中评估分化参数。通过免疫组织化学、酶组织化学和过氧化氢酶比活性研究的过氧化物酶体,根据胎龄(18.5日龄>15.5日龄)和培养类型(球体>三明治>单层)分化得更好。在所有模型中,肝细胞极性标记物ATP、ADP和AMP水解活性均定位错误,位于外侧质膜,而多药耐药相关蛋白2(mrp2)抗原始终正确地免疫定位在顶端膜结构域。在培养物中,观察到荧光素(由mrp2介导)正确分泌到胆小管中。胆小管(紧密连接相关蛋白ZO-1的分支、超微结构和免疫定位)在18.5日龄胎肝中比在15.5日龄胎肝中以及在球体>三明治>单层培养物中分化得更好。我们的结果表明过氧化物酶体区室的变化与胆小管结构以及mrp2介导的分泌功能之间存在平行关系。不同的极化特征不一定同时改变,这表明存在不同的调节机制。