Prudovskiĭ I A, Zelenin A V
Tsitologiia. 1978 Aug;20(8):952-6.
The viability of cytochalasin-enucleated L-cells (cytoplasts) was investigated using autoradiography in addition to fluorescence and electron microscopy. L-cells were enucleated according to a modified Prescott's method (thermostabilized centrifugation, low centrifuge acceleration). Relatively mild conditions of enucleation permitted us to prolong the life-span of cytoplasts up to 24 hours. Soon after enucleation, the disaggregation of polyribosomes in cytoplasts takes place, reflecting, apparently, m-RNA degradation. The cytoplasts are shown to be able to live for 1--2 days with a completely stopped protein synthesis. Acridin orange staining and ultrastructural investigation showed lysosomal hypertrophy in senescent cytoplasts. The death of cytoplasts is suggested to be due to disruption of lysosomes.
除了荧光显微镜和电子显微镜外,还使用放射自显影术研究了细胞松弛素去核L细胞(胞质体)的活力。按照改良的普雷斯科特方法(热稳定离心、低离心加速度)对L细胞进行去核。相对温和的去核条件使我们能够将胞质体的寿命延长至24小时。去核后不久,胞质体中的多核糖体发生解聚,这显然反映了mRNA的降解。结果表明,胞质体在蛋白质合成完全停止的情况下能够存活1至2天。吖啶橙染色和超微结构研究显示衰老胞质体中溶酶体肥大。胞质体的死亡被认为是由于溶酶体的破坏。