Xiao L, Storrie B
Department of Biochemistry and Nutrition, Virginia Polytechnic Institute and State University, Blacksburg 24061.
Exp Cell Res. 1991 Mar;193(1):213-8. doi: 10.1016/0014-4827(91)90558-c.
We have investigated the behavior in in vivo cell fusion experiments of a transitional compartment lying between the endoplasmic reticulum and Golgi apparatus to determine if the compartment, as recognized by the antibody G1/93, might congregate in a similar manner to Golgi apparatus [W. C. Ho et al. (1990) Eur. J. Cell Biol. 52, 315-327]. The distributions of the transitional tubulovesicular compartment, endoplasmic reticulum, and Golgi apparatus in HeLa cells were assessed by immunofluorescent staining using mouse monoclonal antibody G1/93, mouse monoclonal antibody HP 24, and rabbit anti-galactosyltransferase, respectively. In agreement with previous results [W. C. Ho et al. (1990) Eur. J. Cell Biol. 52, 315-327], the Golgi apparatus was observed to congregate gradually over a 3- to 6-h period, forming a large, extended, central Golgi complex in uv-inactivated Sindbis virus-fused HeLa cells. Concomitant with this was a marked congregation of the transitional tubulovesicular compartment. Congregation of the tubulovesicular compartment was not affected by cycloheximide. The endoplasmic reticulum retained its web-like distribution throughout the syncytoplasm and rimmed the nuclear periphery. Treatment of HeLa cells with nocodazole prior to fusion followed by incubation of the syncytia in drug-containing media blocked congregation of the G1/93-positive compartment. With this long-term nocodazole treatment, Golgi apparatus was dispersed into scattered Golgi elements and the G1/93 distribution was endoplasmic reticulum-like. These results suggest that the transitional tubulovesicular compartment recognized by G1/93 is normally structured on microtubules and microtubule organizing centers and may be considered to be a subcompartment of a greater, perinuclear, Golgi complex.
我们研究了位于内质网和高尔基体之间的一个过渡区室在体内细胞融合实验中的行为，以确定由抗体G1/93识别的该区室是否可能以与高尔基体类似的方式聚集[W. C. 何等人（1990年），《欧洲细胞生物学杂志》52卷，315 - 327页]。分别使用小鼠单克隆抗体G1/93、小鼠单克隆抗体HP 24和兔抗半乳糖基转移酶，通过免疫荧光染色评估HeLa细胞中过渡性小管泡区室、内质网和高尔基体的分布。与先前的结果一致[W. C. 何等人（1990年），《欧洲细胞生物学杂志》52卷，315 - 327页]，在紫外线灭活的辛德毕斯病毒融合的HeLa细胞中，观察到高尔基体在3至6小时内逐渐聚集，形成一个大的、延伸的、位于中央的高尔基体复合体。与此同时，过渡性小管泡区室也有明显的聚集。小管泡区室的聚集不受环己酰亚胺的影响。内质网在整个多核细胞质中保持其网状分布，并环绕核周。在融合前用诺考达唑处理HeLa细胞，然后将多核体在含药培养基中孵育，可阻止G1/93阳性区室的聚集。经过这种长期的诺考达唑处理，高尔基体分散成散在的高尔基体元件，G1/93的分布类似内质网。这些结果表明，由G1/93识别的过渡性小管泡区室通常在微管和微管组织中心上构建，并且可以被认为是更大的、核周的高尔基体复合体的一个亚区室。
