Mohri H, Mohri Toshiko, Mabuchi I, Yazaki Ikuko, Sakai H, Ogawa K
Department of Biology, University of Tokyo, Meguro-ku, Tokyo 153.
Department of Biology, Tokyo Metropolitan University, Setagaya-ku, Tokyo 158.
Dev Growth Differ. 1976;18(4):391-398. doi: 10.1111/j.1440-169X.1976.00391.x.
Detection and localization of dynein in cleaving sea urchin eggs were attempted using antidynein serum (prepared against a tryptic fragment of dynein, Fragment A, of sea urchin sperm flagella) and fluorescein conjugated goat antiserum to rabbit γ-globulin. In both unfertilized and newly fertilized eggs, fluorescence was distributed rather uniformly within the cells but was absent from the nuclei. At prophase, intense fluorescence was observed on both sides of nucleus, suggesting accumulation of dynein in developing asters. From metaphase to anaphase, the whole mitotic apparatus (MA) was stained with the exceptions of the chromosomes and pole areas. Fluorescence then again became dispersed within the eggs. Throughout the mitotic process and cytokinesis, the egg cortex including the cleavage furrow was stained intensely, presumably reflecting the presence of dynein in this region. Similar distributions of fluorescence were obtained with the isolated MAs. Neither non-immune serum nor the antiserum to which Fragment A was absorbed stained the eggs. Little staining was obtained with the antiserum against starfish egg myosin. The results, together with the finding that the chromosome motion in the isolated MAs was completely inhibited by anti-dynein serum, but not with the anti-myosin serum, suggest an active role played by a tubulin-dynein system in mitosis.
研究人员尝试利用抗动力蛋白血清(针对海胆精子鞭毛动力蛋白的胰蛋白酶片段A制备)和异硫氰酸荧光素标记的山羊抗兔γ球蛋白血清,对正在分裂的海胆卵中的动力蛋白进行检测和定位。在未受精卵和刚受精的卵中,荧光在细胞内分布较为均匀,但细胞核中没有荧光。在前期,在细胞核两侧观察到强烈的荧光,表明动力蛋白在发育中的星体中积累。从中期到后期,除了染色体和极区外,整个有丝分裂器(MA)都被染色。然后荧光再次在卵内分散。在整个有丝分裂过程和胞质分裂过程中,包括卵裂沟在内的卵皮质被强烈染色,推测这反映了该区域存在动力蛋白。用分离的有丝分裂器也获得了类似的荧光分布。非免疫血清和吸收了片段A的抗血清都没有对卵进行染色。用抗海星卵肌球蛋白的抗血清染色很少。这些结果,连同分离的有丝分裂器中的染色体运动被抗动力蛋白血清完全抑制而抗肌球蛋白血清则没有的发现,表明微管蛋白-动力蛋白系统在有丝分裂中发挥了积极作用。