Cameron I L, Cook K R, Edwards D, Fullerton G D, Schatten G, Schatten H, Zimmerman A M, Zimmerman S
Department of Structural Biology, University of Texas Health Science Center, San Antonio 78284.
J Cell Physiol. 1987 Oct;133(1):14-24. doi: 10.1002/jcp.1041330103.
This study concerned changes in the motional properties of cellular water during the first cell cycle of fertilized sea urchin eggs (Lytechinus variegatus). There was a significant decrease in proton NMR T1 relaxation time and in cytoplasmic ice crystal growth during mitosis and a significant increase in T1 time and cytoplasmic ice crystal size during cleavage. This was not caused by egg water content changes as reflected by egg volume measurements. Removal of both the fertilization membrane and the hyaline layer shortly after fertilization did not alter the pattern of T1 time changes at mitosis and cleavage as compared to whole eggs; thus, the pattern of T1 time changes was attributed to intracellular events. Treatment of fertilized eggs with cytochalasin B, an inhibitor of actin polymerization, did not block the fall in T1 time at mitosis, but did block cytokinesis and the increase in T1 time, which normally occurred at cleavage. A significant pattern of actin disassembly and reassembly at mitosis and cytokinesis was found by studies on the total amount of monomeric actin (G actin) using the DNase I assay. This led to the hypothesis that the observed changes in T1 time and ice crystal size during the first cell cycle were due to the depolymerization and polymerization of cytoplasmic actin. To test this, the effect of the in vitro polymerization of purified actin on the T1 time and on ice crystal growth was examined. It was concluded that changes in the T1 time and ice crystal growth upon polymerization of actin in vitro resembled the changes seen in vivo. These results suggest that changes in the motional properties of cytoplasmic water during the first cell cycle are due, at least in part, to the state of polymerization of cytoplasmic actin.
本研究关注受精海胆卵(多疣荔枝海胆)第一个细胞周期中细胞内水的运动特性变化。在有丝分裂期间,质子核磁共振T1弛豫时间和细胞质冰晶生长显著降低,而在卵裂期间,T1时间和细胞质冰晶大小显著增加。这并非由通过卵体积测量所反映的卵含水量变化引起。受精后不久去除受精膜和透明层,与完整卵相比,在有丝分裂和卵裂时T1时间变化模式并未改变;因此,T1时间变化模式归因于细胞内事件。用细胞松弛素B(一种肌动蛋白聚合抑制剂)处理受精卵,并未阻止有丝分裂时T1时间的下降,但确实阻止了胞质分裂以及通常在卵裂时发生的T1时间增加。通过使用DNase I测定法对单体肌动蛋白(G肌动蛋白)总量进行研究,发现在有丝分裂和胞质分裂时有明显的肌动蛋白解聚和重新组装模式。这导致了一个假设,即第一个细胞周期中观察到的T1时间和冰晶大小变化是由于细胞质肌动蛋白的解聚和聚合。为了验证这一点,研究了纯化肌动蛋白的体外聚合对T1时间和冰晶生长的影响。得出的结论是,肌动蛋白体外聚合时T1时间和冰晶生长的变化类似于体内观察到的变化。这些结果表明,第一个细胞周期中细胞质水运动特性的变化至少部分归因于细胞质肌动蛋白的聚合状态。