Dimitriadis V K
Department of Genetics, Development and Molecular Biology, School of Biology, Aristotle University of Thessaloniki, Greece.
Eur J Histochem. 1993;37(1):65-73.
Immunogold staining of Lowicryl embedded liver sections was used to examine the specific binding of antisera against two pulmonary isozymes (2 and 5) of constitutive form cytochrome P-450, as well as against NADPH-cytochrome P-450 reductase (R) on both untreated and phenobarbital-treated rabbits. In the cytoplasm of liver parenchymal cells, the antisera bound specifically to regions rich in agranular endoplasmic reticulum. In addition, the antibodies bound to plasma membranes in the endothelium of sinusoids and capillaries, as well as to plasma membranes and ground cytoplasm of the cells in bile ducts and interlobular veins. In the liver parenchymal cells, the labelling over the agranular endoplasmic reticulum was uniform throughout the lobule. On the contrary, the labelling over nucleus, mitochondria, granular endoplasmic reticulum and Golgi apparatus was comparable to that observed in control sections. Induction with phenobarbital apparently increased the amount of the agranular endoplasmic reticulum and consequently the total number of gold particles bound in each parenchymal cell compared to controls. In addition, phenobarbital increased the labelling density of isozyme 2 in parenchymal cells, and that of isozymes 2 and 5 in endothelial cells of sinusoids and capillaries.
利用对用Lowicryl包埋的肝脏切片进行免疫金染色,来检测针对组成型细胞色素P - 450的两种肺同工酶(2和5)以及针对NADPH - 细胞色素P - 450还原酶(R)的抗血清,在未处理和苯巴比妥处理的兔子身上的特异性结合情况。在肝实质细胞的细胞质中,抗血清特异性结合富含无颗粒内质网的区域。此外,抗体还结合在肝血窦和毛细血管内皮的质膜上,以及胆管和小叶间静脉细胞的质膜和周围细胞质上。在肝实质细胞中,无颗粒内质网上的标记在整个肝小叶中是均匀的。相反,细胞核、线粒体、颗粒内质网和高尔基体上的标记与对照切片中观察到的情况相当。与对照组相比,苯巴比妥诱导明显增加了无颗粒内质网的量,从而增加了每个实质细胞中结合的金颗粒总数。此外,苯巴比妥增加了实质细胞中同工酶2的标记密度,以及肝血窦和毛细血管内皮细胞中同工酶2和5的标记密度。