Foucault G, Raymond M N, Pudles J
Laboratoire de Structure et Réactivité des Protéines C.N.R.S., U.A. 1131, Université Paris XI, Orsay, France.
Biol Cell. 1987;60(1):63-9. doi: 10.1111/j.1768-322x.1987.tb00547.x.
Unfertilized Paracentrotus lividus egg cytoskeleton is prepared by mild, nonionic detergent extraction at 4 degrees C in buffer systems containing either 2-methyl-2,4-pentanediol (hexylene glycol) or glycerol. These extractions allow the isolation of cytomatrices that maintain the egg form and are 70-80 micron in diameter. DNase inhibition assays show that actin is in polymerized form in these cytomatrices. Ultrastructural observations reveal that the cytoskeletons are made up essentially of 2 categories of filaments, 7-8-nm and 2-4-nm in diameter, respectively. After heavy meromyosin labelling, short, radiating actin filaments are seen in the cortical region, while longer actin filaments are found in the internal region of these cytomatrices. The 2-4-nm filaments of still unknown biochemical nature are organized in a meshwork. In contrast to results found with fertilized eggs, bundles of actin filaments and microtubules are absent; 8-13-nm filaments are not detected.
未受精的紫球海胆卵的细胞骨架是通过在4℃下,在含有2-甲基-2,4-戊二醇(己二醇)或甘油的缓冲系统中,用温和的非离子去污剂提取制备的。这些提取方法能够分离出保持卵形态、直径为70-80微米的细胞基质。脱氧核糖核酸酶抑制试验表明,肌动蛋白在这些细胞基质中呈聚合形式。超微结构观察显示,细胞骨架主要由两类细丝组成,直径分别为7-8纳米和2-4纳米。用重酶解肌球蛋白标记后,在皮质区域可见短的、呈放射状的肌动蛋白细丝,而在这些细胞基质的内部区域发现较长的肌动蛋白细丝。生化性质仍未知的2-4纳米细丝排列成网状结构。与受精卵的结果相反,不存在肌动蛋白细丝束和微管;未检测到8-13纳米的细丝。