Sluder G, Miller F J, Rieder C L
Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545.
Cell Motil Cytoskeleton. 1989;13(4):264-73. doi: 10.1002/cm.970130405.
For animal cells, the relative roles of the centrioles and the pericentriolar material (the centrosomal microtubule organizing center) in controlling the precise doubling of the centrosome before mitosis have not been well defined. To this end we devised an experimental system that allowed us to characterize the capacity of the centrosomal microtubule organizing center to double regularly in the absence of centrioles. Sea urchin eggs were fertilized, stripped of their fertilization envelopes, and fragmented before syngamy. Those activated egg fragments containing just the female pronucleus assembled a monaster at first mitosis. A serial section ultrastructural analysis of such monasters revealed that the radially arrayed microtubules were organized by a hollow fenestrated sphere of electron-dense material, of the same appearance as pericentriolar material, that was devoid of centrioles. We followed individual fragments with only a female pronucleus through at least three cell cycles and found that the monasters did not double between mitoses. The observation that fragments with only a male pronucleus repeatedly divided in a normal fashion indicates that the assembly and behavior of monasters were not artifacts of egg fragmentation. Our results demonstrate that the activity that controls the precise doubling of the centrosome before mitosis is distinct and experimentally separable from the centrosomal microtubule organizing center. Our observations also extend the correlation between the reproductive capacity of a centrosome and the number of centrioles it contains (G Sluder and CL Rieder, 1985a: J. Cell Biol. 100:887-896). For a cell that normally has centrioles, we show that a centrosome without centrioles does not reproduce between mitoses.
对于动物细胞而言,在有丝分裂前,中心粒和中心粒周围物质(即中心体微管组织中心)在控制中心体精确加倍过程中所起的相对作用尚未明确界定。为此,我们设计了一个实验系统,使我们能够在没有中心粒的情况下,对中心体微管组织中心定期加倍的能力进行表征。海胆卵受精后,去除其受精膜,并在配子融合前进行分割。那些仅含有雌原核的活化卵片段在第一次有丝分裂时组装成单星体。对这些单星体进行连续切片超微结构分析发现,呈放射状排列的微管由一个电子致密物质构成的中空有孔球体组织而成,其外观与中心粒周围物质相同,但没有中心粒。我们追踪了仅含有雌原核的单个片段至少三个细胞周期,发现单星体在有丝分裂之间没有加倍。仅含有雄原核的片段能正常反复分裂,这一观察结果表明,单星体的组装和行为并非卵分割的人为产物。我们的结果表明,在有丝分裂前控制中心体精确加倍的活性与中心体微管组织中心是不同的,并且在实验上是可分离的。我们的观察结果还扩展了中心体的繁殖能力与其所含中心粒数量之间的相关性(G·斯卢德和CL·里德,1985a:《细胞生物学杂志》100:887 - 896)。对于正常具有中心粒的细胞,我们表明没有中心粒的中心体在有丝分裂之间不会繁殖。