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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Calmodulin is uniformly distributed during cell division in living stamen hair cells of Tradescantia virginiana
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Calmodulin is uniformly distributed during cell division in living stamen hair cells of Tradescantia virginiana

机译：钙调蛋白在细胞分裂过程中均匀分布在Trade桐的活雄蕊毛细胞中

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Because the activity of calmodulin (CaM) may be dependent dent upon its structural distribution, we have examined its spatial localization in living cells. We have focused on cell division and cell plate formation, where conventional immunofluorescence studies report that CaM is specifically associated with microtubules (MTs) of the spindle and the phragmoplast. In dividing stamen hair cells of Tradescantia virginiana that were injected with fluorescently labeled CaM and examined by confocal laser scanning microscopy (CLSM), we found that the labeled protein is uniformly distributed throughout the cell and is not localized with the phragmoplast MTs or any other obvious structure. To explore why these images from live cells differ from those prepared by immunolabeling, we investigated the fate of CaM during fixation and compared it with the localization of fixable dextran and tubulin. The results show that fixation causes severe changes in cell morphology and in the distribution of CaM and dextran in three quarters of the cells. Conversely, injected rhodamine-tubulin did not show redistribution after fixation. We conclude that in the live cell, CaM is largely uniformly distributed throughout the cytoplasm, and secondly that conventional chemical fixation does not preserve CaM, and probably many other soluble proteins: in its in vivo distribution. The role postulated for CaM in mitosis, solely based on indirect immunofluorescence microscopy, has to be re-evaluated. [References: 56]
机译：由于钙调蛋白（CaM）的活性可能取决于其结构分布，因此我们检查了其在活细胞中的空间定位。我们专注于细胞分裂和细胞板形成，其中传统的免疫荧光研究报告说CaM与纺锤体和原生质膜的微管（MT）特异性相关。在注射荧光标记的CaM并通过共聚焦激光扫描显微镜（CLSM）检查的紫露草的雄蕊毛细胞中，我们发现标记的蛋白均匀地分布在整个细胞中，并且不局限在原生质膜MTs或任何其他明显的结构体。为了探讨为什么这些来自活细胞的图像与通过免疫标记制备的图像不同，我们研究了CaM在固定过程中的命运，并将其与可固定的葡聚糖和微管蛋白的定位进行了比较。结果表明固定会导致四分之三的细胞的细胞形态以及CaM和右旋糖酐的分布发生严重变化。相反，固定后注射的若丹明-微管蛋白未显示出重新分布。我们得出的结论是，在活细胞中，CaM在整个细胞质中大部分均匀地分布，其次，常规化学固定方法无法在体内分布中保留CaM以及许多其他可溶性蛋白质。仅基于间接免疫荧光显微镜，必须重新评估CaM在有丝分裂中的作用。 [参考：56]

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