Brands R, Slot J W, Geuze H J
Eur J Cell Biol. 1983 Nov;32(1):99-107.
In this study, we re-investigated the localization of albumin in hepatocytes using immunocytochemistry (ICC) on ultrathin cryo-sections of rat livers, which were immersion or perfusion-fixed with paraformaldehyde or a mixture of paraformaldehyde and glutaraldehyde. The sections were indirectly labelled with rhodamine-conjugated IgG or with 8 nm protein A-colloidal gold particles. We demonstrate that perfusion-fixation is obligatory for a proper localization of albumin. After perfusion fixation, all hepatocytes of fed and fasted rats were uniformly labelled. At the subcellular level albumin was localized in hepatocytes in the membrane-bound compartments of the entire secretory pathway, i.e. the rough and smooth ER, peripheral Golgi vesicles, all Golgi cisternae and the secretory granules. Concentration of albumin appeared mainly to occur at the cis-Golgi side.
在本研究中,我们利用免疫细胞化学(ICC)技术,在大鼠肝脏超薄冷冻切片上重新研究了白蛋白在肝细胞中的定位。这些切片用多聚甲醛或多聚甲醛与戊二醛的混合物进行浸泡固定或灌注固定。切片用罗丹明偶联的IgG或8纳米蛋白A - 胶体金颗粒进行间接标记。我们证明,灌注固定对于白蛋白的正确定位是必不可少的。灌注固定后,喂食和禁食大鼠的所有肝细胞均被均匀标记。在亚细胞水平上,白蛋白定位于整个分泌途径的膜结合区室中的肝细胞内,即粗面和滑面内质网、周边高尔基体小泡、所有高尔基体潴泡和分泌颗粒。白蛋白的浓缩似乎主要发生在顺面高尔基体一侧。