MICROTUBULE & CENTROSOME ORGANIZATION IN MAMMALIAN CELLS

几个哺乳动物细胞培养系(PtK2，PC 12，HeLa)已经被 用于研究中心体的结构与功能关系 间期和有丝分裂中具有微管组织的材料 采用标准的化学固定和高压冷冻 (最好的朋友)保存猪的超微结构和免疫学细节 中心体、染色体、微管、膜和中间体 使用了几种抗体并对其进行了传播分析 以及扫描电子和免疫荧光显微镜 利用抗中心体的人自身抗体(SPJ)， 小鼠单锥体。抗中间体微管蛋白(M)抗体 抗波形蛋白细丝抗体(Ah6)，DAR染色DNA。 在对照细胞的间期，中心体物质紧密 通过纤维网络与核膜相连，并成为 在有丝分裂期间逐渐解离，在那里它的功能是 基于微管的有丝分裂的微管组织中心 仪器。与海胆卵和胚胎不同，在海胆卵和胚胎中有微管 是细胞周期特定的扩增和压缩所必需的 中心体(Schatten等人，1988，Cell Motil.赛托斯克尔。11、 248-259)，微管不是细胞周期特异性所必需的 哺乳动物细胞中中心体的研究进展。这些研究扩展了 根据以前的发现(Joswig和Petzelt，1990，Cell Motil)。赛托斯克尔。 15,181-192)，并支持不同的机制 中心体膨胀和压缩行为用于不同的 真核物种。通过使用高压冷冻，这个项目是 可能有助于我们对折叠机制的理解 和中心体物质的展开，也将有助于我们的 对中心体-细胞骨架相互作用的认识 受精、细胞分裂、细胞分化和胚胎 发展。SCXENTXFXC子项目批准号：P41RR00570-27

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