Villanueva Marco A, Schindler Melvin, Wang John L
Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, U.N.A.M., Apartado Postal 510-3, Cuernavaca, Morelos 62250, Mexico.
Cell Biol Int. 2005 Nov;29(11):936-42. doi: 10.1016/j.cellbi.2005.08.005. Epub 2005 Nov 2.
The microfilament network of cultured Glycine max cells (SB-1 line), and protoplasts was visualized with rhodamine-phalloidin under conditions that lysed the protoplast and changed the cell shape. The whole cell had the typical microfilament distribution of a "cage" around the nucleus, from which the large subcortical cables and transvacuolar strands radiated towards the cortex until it reached the cortical microfilament network. Upon cell wall removal, the network conserved its compartmentalization. Thus, the redistribution of the shape where the vacuole becomes a central entity, made the cytoplasm displace peripherally, but the network distribution was conserved. When protoplasts were lysed in a low osmotic medium, the vacuoles were gradually released intact. Under these conditions, the F-actin staining remained within the ghost of the cell, but none was detected in either emerging or almost completely released vacuoles. Most of the released F-actin was found in debris from the cell lysate in the form of microfilaments. When the ghosts were constrained in a coverslip with an air bubble, the shape of the ghost changed accordingly, but the microfilament network distribution remained constant. These results provide further evidence that the vacuole of plant cells does not have detectable associated F-actin. In addition, we demonstrate that the actin microfilament network is a moldable entity that can change its shape but keeps its distribution under constant conditions, in these cultured cells.
在使原生质体裂解并改变细胞形状的条件下,用罗丹明 - 鬼笔环肽对培养的大豆细胞(SB - 1系)和原生质体的微丝网络进行可视化观察。整个细胞具有围绕细胞核的典型“笼状”微丝分布,从该分布中,大的皮层下索和跨液泡丝向皮层辐射,直至到达皮层微丝网络。去除细胞壁后,该网络保持其分区状态。因此,液泡成为中心实体的形状重新分布,使细胞质向周边位移,但网络分布得以保留。当原生质体在低渗介质中裂解时,液泡逐渐完整地释放出来。在这些条件下,F - 肌动蛋白染色保留在细胞的“幽灵”内,但在刚出现或几乎完全释放的液泡中均未检测到。大部分释放的F - 肌动蛋白以微丝的形式存在于细胞裂解物的碎片中。当“幽灵”被气泡限制在盖玻片上时,“幽灵”的形状相应改变,但微丝网络分布保持不变。这些结果进一步证明植物细胞的液泡没有可检测到的相关F - 肌动蛋白。此外,我们证明在这些培养细胞中,肌动蛋白微丝网络是一个可塑造的实体,它可以改变形状,但在恒定条件下保持其分布。