Mizuno M, Singer S J
Department of Biology, University of California at San Diego, La Jolla 92037.
J Cell Sci. 1994 May;107 ( Pt 5):1321-31. doi: 10.1242/jcs.107.5.1321.
The intracellular transport of secretory proteins involves at an early stage the formation of vesicles from transitional elements of the endoplasmic reticulum (ER) containing these proteins and the transfer of these vesicles to the cis-face of the Golgi apparatus. We propose that the latter transfer process does not occur by random diffusion, but is instead mediated by tracking along stable microtubules. To test this proposal, we have carried out double immunoelectron microscopic labeling experiments on frozen sections of HepG2 hepatoma cells secreting the protein human serum albumin (HSA). By a cycloheximide treatment protocol, the stage during which the transfer of newly synthesized HSA from the ER to the Golgi apparatus occurs in vivo was determined. Sections of the cells were then double immunolabeled using primary antibodies to HSA and to glu-tubulin, the latter specifically detecting stable microtubules. We observed a significantly high frequency of HSA-containing structures between the ER and the Golgi apparatus with which stable microtubules were closely associated. These results support the proposal that stable microtubules may play a critical role in directing the transfer process from the ER to the Golgi apparatus.
分泌蛋白的细胞内运输在早期涉及从含有这些蛋白质的内质网(ER)的过渡元件形成囊泡,并将这些囊泡转移到高尔基体的顺面。我们提出,后一种转移过程不是通过随机扩散发生的,而是通过沿着稳定的微管追踪来介导的。为了验证这一观点,我们对分泌人血清白蛋白(HSA)的HepG2肝癌细胞的冷冻切片进行了双重免疫电子显微镜标记实验。通过环己酰亚胺处理方案,确定了体内新合成的HSA从内质网转移到高尔基体的阶段。然后使用针对HSA和γ-微管蛋白的一抗对细胞切片进行双重免疫标记,后者特异性检测稳定的微管。我们观察到在内质网和高尔基体之间,含有HSA的结构与稳定微管紧密相关的频率显著较高。这些结果支持了稳定微管可能在指导从内质网到高尔基体的转移过程中起关键作用的观点。