Asada S, Yagura T
Department of Chemistry, Kwansei Gakuin University, Hyogo, Japan.
Cell Struct Funct. 1995 Dec;20(6):445-53. doi: 10.1247/csf.20.445.
A monoclonal antibody (mAbG3A5) recognizing p138 antigen was used to identify the Golgi cisternal membrane and determine behavior of Golgi fragments during mitosis in HeLa cells. At the start of mitosis, Golgi stacks identified with the mAbG3A5 antibody were fragmented into fine membrane vesicles which were distributed throughout the cytoplasm leaving only the region of the chromosome cluster unoccupied. On Western immunoblotting analysis, p138 was found associated with the membrane fraction prepared from mitotic HeLa cells having a buoyant density the same as that of interphase Golgi membranes. In addition to the fine membrane vesicles, clusters labeled with mAbG3A5 antibody were frequently observed in mitotic cells. They numbered 11 on average per mitotic cell and consisted of fine membrane vesicles of which membrane region was labeled with the mAbG3A5 antibody. This fact indicates that the membrane vesicles in mitotic Golgi clusters were also part of the fragments of Golgi cisternae. The number of mitotic Golgi clusters per mitotic cell was constant from prophase to anaphase, increasing twofold at telophase, although the average size of mitotic Golgi cluster remained unchanged throughout mitosis. The increase in number of mitotic Golgi clusters at telophase was accompanied by decrease in immunofluorescence of fine membrane vesicles. Treatment with nocodazole caused the disappearance of the mitotic Golgi clusters from prophase cells; however upon removal of it, they were reformed. These results suggest that during mitosis the Golgi apparatus were fragmented to fine membrane vesicles leaving only a part as mitotic Golgi clusters and were reassembled through tentative clustering of the fine membrane vesicles at the end of mitosis.
一种识别p138抗原的单克隆抗体（mAbG3A5）被用于鉴定高尔基体潴泡膜，并确定HeLa细胞有丝分裂期间高尔基体片段的行为。在有丝分裂开始时，用mAbG3A5抗体鉴定的高尔基体堆叠片段化为细小的膜泡，这些膜泡分布在整个细胞质中，仅染色体簇区域未被占据。在蛋白质免疫印迹分析中，发现p138与从有丝分裂HeLa细胞制备的膜组分相关，其浮力密度与间期高尔基体膜相同。除了细小的膜泡外，在有丝分裂细胞中还经常观察到用mAbG3A5抗体标记的簇。每个有丝分裂细胞平均有11个，由细小的膜泡组成，其膜区域用mAbG3A5抗体标记。这一事实表明，有丝分裂高尔基体簇中的膜泡也是高尔基体潴泡片段的一部分。从前期到后期，每个有丝分裂细胞中分裂期高尔基体簇的数量是恒定的，末期增加两倍，尽管有丝分裂高尔基体簇的平均大小在整个有丝分裂过程中保持不变。末期有丝分裂高尔基体簇数量的增加伴随着细小膜泡免疫荧光的降低。用诺考达唑处理导致前期细胞中分裂期高尔基体簇消失；然而，去除诺考达唑后，它们又重新形成。这些结果表明，在有丝分裂期间，高尔基体破碎成细小的膜泡，仅留下一部分作为有丝分裂高尔基体簇，并在有丝分裂末期通过细小膜泡的暂时聚集而重新组装。
