Schatten H, Schatten G, Mazia D, Balczon R, Simerly C
Proc Natl Acad Sci U S A. 1986 Jan;83(1):105-9. doi: 10.1073/pnas.83.1.105.
The forms and locations of centrosomes in mouse oocytes and in sea urchin eggs were followed through the whole course of fertilization and first cleavage by immunofluorescence microscopy. Centrosomes were identified with an autoimmune antiserum to centrosomal material. Staining of the same preparations with tubulin antibody and with the DNA dye Hoechst 33258 allowed the correlation of the forms of the centrosomes with the microtubule structures that they generate and with the stages of meiosis, syngamy, and mitosis. The results with sea urchin eggs conform to Boveri's view on the paternal origin of the functional centrosomes. Centrosomes are seen in spermatozoa and enter the egg at fertilization. Initially, the centrosomes are compact, but as the eggs enter the mitotic cycle the forms of the centrosomes go through a cycle in which they spread during interphase, apparently divide, and condense into two compact poles by metaphase. In anaphase, they spread to form flat poles. In telophase and during reconstitution of the daughter nuclei, the centrosomal material is disposed as hemispherical caps around the poleward surfaces of the nuclei. Mouse sperm lack centrosomal antigen. In the unfertilized mouse oocyte, the meiotic spindle poles are displayed as broad-beaded centrosomes. In addition, centrosomal material is detected in the cytoplasm as particles, about 16 in number, which are foci of small aster-like arrays of microtubules. The length and number of astral microtubules correlate with the size of the centrosomal foci. After sperm incorporation, as the pronuclei develop and more cytoplasmic microtubules assemble, a few of the foci associate with the peripheries of the nuclei. The number of foci multiplies during the first cell cycle. At the end of interphase, all of the centrosomal foci have concentrated on the nuclear peripheries and the cytoplasmic microtubules have disappeared. At prophase, the centrosomes are seen as two irregular clusters, marking the poles which, at metaphase and anaphase, appear as rough bands with foci, and the spindle is typically barrel-shaped. At telophase, the centrosomes are seen as arcs that lie on the nuclear peripheries after cleavage. The ordering of microtubules in all the stages reflects the shapes of the centrosomes. The findings on the sea urchin confirm the classical theory of the paternal origin of centrosomes and contrast with observations tracing the mitotic poles of the mouse egg to maternal centrosomal material. This evidence strengthens the conclusion that mouse centrosomes derive from the oocyte.
通过免疫荧光显微镜观察，追踪了小鼠卵母细胞和海胆卵中中心体在受精和第一次卵裂全过程中的形态和位置。用针对中心体物质的自身免疫抗血清鉴定中心体。用微管蛋白抗体和DNA染料Hoechst 33258对相同的标本进行染色，从而能够将中心体的形态与它们所产生的微管结构以及减数分裂、受精和有丝分裂的阶段联系起来进行研究。海胆卵的实验结果符合博韦里关于功能性中心体父系起源的观点。在精子中可以看到中心体，受精时进入卵子。最初，中心体是紧密的，但随着卵子进入有丝分裂周期，中心体的形态经历一个循环，在间期它们扩散，显然会分裂，到中期浓缩成两个紧密的极。在后期，它们扩散形成扁平的极。在末期以及子核重建过程中，中心体物质呈半球形帽状分布在核向极表面周围。小鼠精子缺乏中心体抗原。在未受精的小鼠卵母细胞中，减数分裂纺锤体极表现为宽珠状中心体。此外，在细胞质中检测到中心体物质呈颗粒状，数量约为16个，它们是小星状微管阵列的焦点。星状微管的长度和数量与中心体焦点的大小相关。精子入卵后，随着原核发育以及更多细胞质微管组装，一些焦点与核周边相连。在第一个细胞周期中焦点数量增加。在间期结束时，所有中心体焦点都集中在核周边，细胞质微管消失。在前期，中心体表现为两个不规则的簇，标志着两极，在中期和后期，两极表现为有焦点的粗带，纺锤体通常呈桶状。在末期，中心体表现为弧形，在卵裂后位于核周边。所有阶段微管的排列都反映了中心体的形状。海胆的研究结果证实了中心体父系起源的经典理论，与将小鼠卵有丝分裂极追溯到母系中心体物质的观察结果形成对比。这一证据强化了小鼠中心体源自卵母细胞的结论。
