MICROTUBULE & CENTROSOME ORGANIZATION IN MAMMALIAN CELLS

几种哺乳动物细胞培养系（PtK 2、PC 12、HeLa）已经被研究。 用于研究中心体的结构-功能关系 在分裂间期和有丝分裂期间具有微管组织的物质 采用标准的化学固定以及高压冷冻 （BFF）。 为了保留细胞的超微结构和免疫学细节， 中心体、染色体、微管、膜和中间体 使用了几种抗体，并通过透射电镜进行了分析。 扫描电子和免疫荧光显微镜 利用针对中心体（SPJ）的人自身免疫抗体， 小鼠单锥。 微管蛋白（M）的抗体， 抗波形蛋白（Ah-6）的细丝抗体和用于染色DNA的DAR。 在对照细胞的分裂间期，中心体物质紧密结合， 通过纤维网络与核膜相连， 在有丝分裂过程中逐渐分离， 微管有丝分裂的微管组织中心 设备. 不像海胆卵和胚胎中的微管 是细胞周期特异性扩增和压缩 中心体（Schatten等人，1988年，细胞运动。 细胞骨架 11, 248-259），微管对于细胞周期特异性的 哺乳动物细胞中中心体的进展。 这些研究扩展了 基于先前的发现（Joswig和Petzelt，1990，Cell Motil. 细胞骨架 15，181-192），并支持这一概念，不同的机制， 中心体扩展和压缩行为被用于不同的 真核生物 本工程采用高压冻结法， 可能有助于我们理解折叠的机制 和展开的中心体材料，也将有助于我们的 理解中心体-细胞骨架的相互作用 受精、细胞分裂、细胞分化和胚胎 发展 SCXENTXFXC子项目批准编号：P41 RR 00570 -27

Several mammalian cell culture lines (PtK2, PC 12, HeLa) have been utilized to study the structure-function relationship of centrosomal material with microtubule organization during interphase and mitosis employing standard chemical fixation as well as high pressure freezing (BFF). To preserve ultrastructural and immunological details of centrosomes, chromosomes, microtubules, membranes and intermediate filaments several antibodies were used and analyzed with transmission and scanning electron as well as with immunofluorescence microscopy utilizing a human autoinunune antibody against centrosomes (SPJ), a mouse monoconal. antibody against tubulin (M), an intermediate filament antibody against vimentin (Ah-6), and DAR to stain DNA. During interphase in control cells, centrosomal material is closely associated with the nuclear envelope by a fibrous network and becomes gradually dissociated during mitosis where it functions as the microtubule organizing center for the microtubule-based mitotic apparatus. Unlike in sea urchin eggs and embryos where microtubules are needed for cell-cycle specific expansion and compaction of centrosomes (Schatten et al., 1988, Cell Motil. Cytoskel. 11, 248-259), microtubules are not required for cell-cycle specific progression of centrosomes in mammalian cells. These studies extend on previous findings (Joswig and Petzelt, 1990, Cell Motil. Cytoskel. 15, 181-192) and support the notion that different mechanisms for centrosome expansion and compaction behavior are used in different eukaryotic species. By using high pressure freezing, this project is likely to contribute to our understanding on the mechanisms of folding and unfolding of centrosomal material and will also contribute to our understanding on centrosome-cytoskeletal interactions during fertilization, cell division, cell differentiation, and embryo development. SCXENTXFXC SUBPROJECT GRANT NUMBER: P41RR00570-27