Keller G A, Glass C, Louvard D, Steinberg D, Singer S J
J Histochem Cytochem. 1986 Sep;34(9):1223-30. doi: 10.1177/34.9.3525668.
Synthesis and intracellular transport of two secretory proteins, serum albumin (SA) and apolipoprotein B (apo B) have been synchronized in primary cultures of normal rat hepatocytes to make possible immunocytochemical study of the transport pathway. Under appropriate conditions of cycloheximide treatment, synthesis of new protein was inhibited and, by double immunofluorescent labeling, the cells were found to be largely depleted of the SA and apo B previously synthesized. Re-initiation of protein synthesis led to sequential appearance of SA and apo B, first in the endoplasmic reticulum, then in the Golgi complex, and finally at the cell surface. These results indicate that it should be feasible to use this cell system for high-resolution investigation of the sequence of structures involved in intracellular transport of SA and apo B by corresponding immunolabeling experiments as observed by electron microscopy.
在正常大鼠肝细胞原代培养物中，已使两种分泌蛋白，即血清白蛋白（SA）和载脂蛋白B（apo B）的合成与细胞内运输同步，从而使对运输途径进行免疫细胞化学研究成为可能。在适当的环己酰亚胺处理条件下，新蛋白质的合成受到抑制，通过双重免疫荧光标记发现，细胞中先前合成的SA和apo B大量减少。蛋白质合成的重新启动导致SA和apo B依次出现，首先在内质网中，然后在高尔基体中，最后出现在细胞表面。这些结果表明，通过电子显微镜观察相应的免疫标记实验，利用该细胞系统对SA和apo B细胞内运输所涉及的结构序列进行高分辨率研究应该是可行的。
