Pypaert M, Nilsson T, Berger E G, Warren G
Cell Biology Laboratory, Imperial Cancer Research Fund, London, UK.
J Cell Sci. 1993 Mar;104 ( Pt 3):811-8. doi: 10.1242/jcs.104.3.811.
HeLa cells were incubated with 15 nm BSA-gold for 1 or 2 hours to mark the endocytic pathway and mitotic cells were then isolated by shake-off. Thin, frozen sections were labelled with antibodies against two resident Golgi markers, beta-(1,4)-galactosyltransferase and N-acetylglucosaminyltransferase I. Detection of the latter was aided by the use of a HeLa cell line stably expressing a myc-tagged version of the endogenous protein. The secondary antibodies were coupled to either 5 or 10 nm gold so that the distribution of each of the three markers could be followed. Qualitative and quantitative studies showed that there were two populations of clusters, those described by us earlier and termed Golgi clusters (Lucocq et al. (1987) J. Cell Biol. 104, 865-874), containing either or both Golgi markers, and clusters of tubular endosomes containing BSA-gold. There was very little overlap showing that Golgi clusters cannot be tubular endosomes as concluded by Tooze and Hollinshead (1992) Eur. J. Cell Biol. 58, 228-242.
将HeLa细胞与15纳米的牛血清白蛋白-金孵育1或2小时以标记内吞途径，然后通过振荡分离有丝分裂细胞。薄的冰冻切片用针对两种高尔基体内驻留标记物β-(1,4)-半乳糖基转移酶和N-乙酰葡糖胺基转移酶I的抗体进行标记。通过使用稳定表达内源性蛋白的myc标签版本的HeLa细胞系来辅助检测后者。二抗与5或10纳米的金偶联，以便追踪三种标记物中每一种的分布。定性和定量研究表明，存在两种簇群，即我们之前描述的那些并称为高尔基体簇（Lucocq等人，(1987)《细胞生物学杂志》104卷，865 - 874页），其包含一种或两种高尔基体标记物，以及含有牛血清白蛋白-金的管状内体簇。几乎没有重叠，这表明高尔基体簇不可能是如Tooze和Hollinshead（1992年，《欧洲细胞生物学杂志》58卷，228 - 242页）所推断的管状内体。
