Comparison of F-actin fluorescent labeling methods in pollen tubes of Lilium davidii

Comparison of F-actin fluorescent labeling methods in pollen tubes of Lilium davidii
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百合花粉管F-actin荧光标记方法比较

DOI:
10.1007/s00299-005-0935-y
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发表时间:
2005-03
期刊:
影响因子:
6.2
通讯作者:
Y. Li
Y. Li
中科院分区:
生物学2区
文献类型:
--
作者:
L. Wang;Y-M. Liu;Y. Li

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参考文献

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用不同的方法对花粉管中的F-肌动蛋白进行荧光标记,在文献中产生了不一致的结果。在此,我们报道了过去一直用于固定缓冲液中的EGTA,它被认为有助于细胞骨架的稳定,它可以显著影响F-肌动蛋白的分布,并导致在花粉管顶端形成厚厚的F-肌动蛋白束。我们还发现,在花粉管固定过程中,前5min的真空渗透可以更好地保存正常的细胞质结构和F-肌动蛋白的分布。相反,间马来酰亚胺苯甲酸酯(MBS)在化学固定前处理导致花粉管顶端粗大的F-肌动蛋白束游离区缩短。综上所述,我们的结果表明,从固定缓冲液中排除EGTA和MBS,结合固定前5min的真空渗透,可以提高百合花粉管中F-肌动蛋白的荧光标记。
Fluorescence labeling of F-actin in pollen tubes by various methods has produced inconsistent results in the literature. Here, we report that EGTA, which was always used in fixative buffers in the past and thought to help cytoskeleton stabilization, can significantly affect F-actin distribution and lead to the formation of thick F-actin bundles at the tip of the pollen tube. We also found that vacuum-infiltration for the first 5 min during pollen tube fixation can better preserve normal cytoplasm structure and F-actin distribution. In contrast, m-maleimidobenzoic acid N-hydroxysuccinimide ester (MBS) treatment before chemical fixation resulted in a shortening of the free zone of thick F-actin bundles in the pollen tube tip. Taken together, our results suggest that exclusion of EGTA and MBS from the fixative buffer, in combination with vacuum-infiltration in the first 5 min of fixation, can improve F-actin fluorescence labeling in pollen tubes of Lilium davidii.
DOI: --
发表时间: 1998-10
期刊: Acta Botanica Sinica
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