Heidemann S R, Kirschner M W
J Cell Biol. 1975 Oct;67(1):105-17. doi: 10.1083/jcb.67.1.105.
We have assayed various materials for their ability to induce aster formation by microinjection into unfertilized eggs of Xenopus laevis. We have found that purified basal bodies from Chlamydomonas reinhardtii and Tetrahymena pyriformis induce the formation of asters and irregular cleavage furrows within 1 h after injection. Other microtubule structures such as flagella, flagellar axonemes, cilia, and brain microtubules are completely ineffective at inducing asters or cleavage furrows in unfertilized eggs. When known amounts of sonicated Tetrahymena and Chlamydomonas preparations are injected into unfertilized eggs, 50% of the injected eggs show a furrowing response at approximately 3 cell equvalents for Chlamydomonas and 0.1 cell equivalent for Tetrahymena. These results are close to those expected if basal bodies were the effective astral-inducing agent in these cells. Other materials effective at inducing asters in unfertilized eggs, such as crude brain nuclei, sperm, and a particulate fraction from brain known to induce parthenogenesis in eggs of Rana pipiens, probably contain centrioles as the effective agent. Our experiments provide the first functional assay to indicate that centrioles play an active role in aster initiation. None of the injected materials effective in unfertilized eggs produced any observable response in fully grown oocytes. Oocytes and eggs were found to have equal tubulin pools as judged by colchicine-binding activity. Therefore, the inability of oocytes to form asters cannot be due to a lack of an organizing center or to a lack of tubulin. Experiments in which D2O was found to stimulate aster-like fibrous areas in eggs but not oocytes suggest that the inability of oocytes to form asters may be due to an inability of tubulin in oocytes to assemble.
我们通过将各种物质显微注射到非洲爪蟾未受精卵中,检测了它们诱导星体形成的能力。我们发现,从莱茵衣藻和梨形四膜虫中纯化得到的基体,在注射后1小时内可诱导星体和不规则分裂沟的形成。其他微管结构,如鞭毛、鞭毛轴丝、纤毛和脑微管,在未受精卵中完全无法诱导星体或分裂沟的形成。当将已知量的经超声处理的四膜虫和衣藻制剂注射到未受精卵中时,约50%的注射卵在衣藻约3个细胞当量、四膜虫0.1个细胞当量时出现沟裂反应。这些结果与如果基体是这些细胞中有效的星体诱导剂时预期的结果相近。其他在未受精卵中有效诱导星体形成的物质,如粗脑细胞核、精子以及已知能诱导牛蛙卵孤雌生殖的脑微粒部分,可能含有中心粒作为有效成分。我们的实验提供了首个功能检测,表明中心粒在星体起始过程中发挥积极作用。在完全成熟的卵母细胞中,没有一种注射到未受精卵中有效的物质产生任何可观察到的反应。通过秋水仙素结合活性判断,发现卵母细胞和卵具有相等的微管蛋白库。因此,卵母细胞无法形成星体不是由于缺乏组织中心或缺乏微管蛋白。重水被发现能刺激卵中类似星体的纤维区域但不能刺激卵母细胞的实验表明,卵母细胞无法形成星体可能是由于卵母细胞中的微管蛋白无法组装。