Dahan S, Ahluwalia J P, Wong L, Posner B I, Bergeron J J
Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada.
J Cell Biol. 1994 Dec;127(6 Pt 2):1859-69. doi: 10.1083/jcb.127.6.1859.
The intrahepatic distribution of apolipoprotein E has been assessed by immunogold labeling of cryosections as well as by Western blotting of organelles isolated from liver homogenates. Both techniques supported the prior analytical fractionation studies of Wong (1989) who concluded that intrahepatic apoE was largely endosomal. All endosomal components decorated by gold particles indicative of apoE antigenicity in cryosections appeared filled with lipoprotein-like particles thereby accounting for this prominent morphological feature of isolated liver endosomes. The distribution of gold particles about the hepatic Golgi apparatus revealed a high content of apoE in closely apposed endosomes, ca. 400 nm in diameter, double labeled for apoE and internalized HRP. Remarkably, apoE (but not internalized HRP) was also observed within saccular distensions of all saccules of stacked Golgi cisternae but absent from the flattened saccular components as was also observed for apoB. This contrasted with albumin, the major secretory protein, which was uniformly distributed throughout the hepatic Golgi apparatus. These observations support a growing body of evidence for intra-Golgi sorting of secretory material in hepatic Golgi apparatus. The lack of any immunoreactive apoE or albumin in small 70-90 nm vesicles about the Golgi cisternae suggests limits to current models of vesicle-mediated intra-Golgi transport.
载脂蛋白E在肝内的分布已通过冷冻切片的免疫金标记以及从肝脏匀浆中分离的细胞器的蛋白质免疫印迹法进行了评估。这两种技术都支持了Wong(1989年)先前的分析分级研究,他得出结论,肝内载脂蛋白E主要存在于内体中。冷冻切片中所有被金颗粒标记(表明具有载脂蛋白E抗原性)的内体成分似乎都充满了脂蛋白样颗粒,这就解释了分离出的肝脏内体这一显著的形态学特征。围绕肝高尔基体的金颗粒分布显示,在紧密相邻的内体中载脂蛋白E含量很高,这些内体直径约400 nm,同时被载脂蛋白E和内化的辣根过氧化物酶双标记。值得注意的是,在堆叠的高尔基池所有囊泡的囊状扩张内也观察到了载脂蛋白E(但没有内化的辣根过氧化物酶),而在扁平的囊状成分中则没有,载脂蛋白B也有同样的情况。这与主要分泌蛋白白蛋白形成对比,白蛋白在整个肝高尔基体中均匀分布。这些观察结果支持了越来越多的证据,表明肝高尔基体中存在分泌物质的高尔基体内分选。在高尔基池周围70 - 90 nm的小囊泡中缺乏任何免疫反应性的载脂蛋白E或白蛋白,这表明目前囊泡介导的高尔基体内运输模型存在局限性。