An actin network is present in the cytoplasm throughout the cell cycle of carrot cells and associates with the dividing nucleus.
An actin network is present in the cytoplasm throughout the cell cycle of carrot cells and associates with the dividing nucleus.
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DOI：
10.1083/jcb.105.1.387
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发表时间：
1987-07
期刊：
影响因子：
--
通讯作者：
Lloyd CW
中科院分区：
文献类型：
--
作者：
Traas JA;Doonan JH;Rawlins DJ;Shaw PJ;Watts J;Lloyd CW
采用罗丹明赖氨酸phallotoxin （RLP）对胡萝卜循环悬浮培养细胞中的f -肌动蛋白网络进行了研究。除了常规的甲醛固定外，在加入RLP之前，我们还使用了两种不同的不固定方法：在稳定缓冲液中提取细胞；用直流电脉冲（电穿孔）在质膜上诱导瞬时孔隙。这些引入RLP的替代方法揭示了在醛固定细胞中未见的肌动蛋白网络的其他特征。该网络在非扁平细胞中的三维组织通过投影立体对来证明，这些立体对来自于计算机增强图像的穿过焦点系列。f -肌动蛋白以四种相互连接的结构存在于间期细胞中：围绕细胞核的网状结构；穿过液泡链的粗索和深入细胞质；皮层细胞质内较细的束状网络；甚至更细的细丝在细胞周围有序地横向排列。肌动蛋白网络在分裂过程中组织方式不同，但它不像皮层微管那样消失。当染色质开始凝聚时，RLP染色中央丝状皮质带（前期）；它会染色有丝分裂纺锤体（最近由Seagull等人发现）[Seagull， R. W.， M. Falconer, and C. A. Weerdenburg, 1987， J.细胞生物学]。（如Clayton， L., and c.w. Lloyd, 1985, Exp. Cell Res., 156:231-238）。然而，现在的研究表明，在整个分裂过程中，细胞质中持续存在一个额外的f -肌动蛋白网络，依次与前期带、有丝分裂纺锤体和细胞动力学的膜质体相关。
We have studied the F-actin network in cycling suspension culture cells of carrot (Daucus carota L.) using rhodaminyl lysine phallotoxin (RLP). In addition to conventional fixation with formaldehyde, we have used two different nonfixation methods before adding RLP: extracting cells in a stabilizing buffer; inducing transient pores in the plasma membrane with pulses of direct current (electroporation). These alternative methods for introducing RLP revealed additional features of the actin network not seen in aldehyde-fixed cells. The three- dimensional organization of this network in nonflattened cells was demonstrated by projecting stereopairs derived from through-focal series of computer-enhanced images. F-actin is present in interphase cells in four interconnected configurations: a meshwork surrounding the nucleus; thick cables in transvacuolar strands and deep in the cytoplasm; a finer network of bundles within the cortical cytoplasm; even finer filaments that run in ordered transverse array around the cell periphery. The actin network is organized differently during division but it does not disappear as do the cortical microtubules. RLP stains a central filamentous cortical band as the chromatin begins to condense (preprophase); it stains the mitotic spindle (as recently shown by Seagull et al. [Seagull, R. W., M. Falconer, and C. A. Weerdenburg, 1987, J. Cell Biol., 104:995-1004] for aldehyde fixed suspension cells) and the cytokinetic apparatus (as shown by Clayton, L., and C. W. Lloyd, 1985, Exp. Cell Res., 156:231-238). However, it is now shown that an additional network of F-actin persists in the cytoplasm throughout division associating in turn with the preprophase band, the mitotic spindle, and the cytokinetic phragmoplast.