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向活的培养细胞中显微注射钒酸盐会抑制纤毛运动而非跳跃运动。

Ciliary but not saltatory movements are inhibited by vanadate microinjected into living cultured cells.

作者信息

Buckley I, Stewart M

出版信息

Cell Motil. 1983;3(2):167-84. doi: 10.1002/cm.970030206.

Abstract

To test the idea that saltatory organelle movements of nonmuscle cells might be driven by microtubule-dynein interactions, we microinjected vanadate into several different types of cultured cell. Solutions of sodium metavanadate made up in a simple buffered salt solution were pressure microinjected into fully spread cells in an open-topped culture chamber placed on the stage of an inverted microscope. The cells were observed by oil-immersion phase-contrast optics and results were recorded on movie film. Vanadate, at 10(-5)-10(-2) M, microinjected into cultured chick embryo fibroblasts, failed to inhibit organelle movements. To test the effectiveness of vanadate's inhibitory action under living cell conditions, ciliated epithelial cells were microinjected. In these cells even the smallest microinjection of 5 X 10(-5) M vanadate caused an immediate cessation of ciliary beating. Moreover, in cells that were well spread it was found that whereas vanadate, at 5 X 10(-5)-5 X 10(-3) M, inhibited ciliary motion, it failed to inhibit organelle saltations in the same cell. To determine whether vanadate would inhibit a living actin-myosin system, myocardial cells were also microinjected. Following microinjection of 5 X 10(-5) and 5 X 10(-4) M vanadate a temporary tonic contraction (which also occurred following microinjection of buffer alone) was followed by regular beating. Taken together these results demonstrate that in living cell systems microtubule-dynein interactions are as sensitive to vanadate inhibition as they are in demembranated model systems, and that a working actin-myosin system in a living muscle cell does not share this great sensitivity. In light of the pronounced differential inhibitory effects of vanadate on the movements of cilia and organelles, our results suggest that saltatory organelle movements in chick embryo fibroblasts and rabbit oviduct epithelial cells are unlikely to be brought about by microtubule-dynein interactions.

摘要

为了验证非肌肉细胞的跳跃式细胞器运动可能由微管-动力蛋白相互作用驱动这一想法,我们将钒酸盐显微注射到几种不同类型的培养细胞中。用简单的缓冲盐溶液配制的偏钒酸钠溶液通过压力显微注射到倒置显微镜载物台上敞口培养室中完全铺展的细胞内。通过油浸相差光学显微镜观察细胞,并将结果记录在电影胶片上。显微注射到培养的鸡胚成纤维细胞中的10⁻⁵ - 10⁻² M钒酸盐未能抑制细胞器运动。为了测试钒酸盐在活细胞条件下抑制作用的有效性,对纤毛上皮细胞进行了显微注射。在这些细胞中,即使是最小剂量5×10⁻⁵ M的钒酸盐显微注射也会立即导致纤毛跳动停止。此外,在铺展良好的细胞中发现,5×10⁻⁵ - 5×10⁻³ M的钒酸盐虽然抑制了纤毛运动,但未能抑制同一细胞中的细胞器跳跃。为了确定钒酸盐是否会抑制活的肌动蛋白-肌球蛋白系统,也对心肌细胞进行了显微注射。显微注射5×10⁻⁵和5×10⁻⁴ M钒酸盐后,先是出现短暂的强直性收缩(单独显微注射缓冲液时也会出现),随后是有规律的跳动。综合这些结果表明,在活细胞系统中,微管-动力蛋白相互作用对钒酸盐抑制的敏感性与在去膜模型系统中一样,而活肌肉细胞中的功能性肌动蛋白-肌球蛋白系统并不具有这种高度敏感性。鉴于钒酸盐对纤毛和细胞器运动具有明显的差异抑制作用,我们的结果表明,鸡胚成纤维细胞和兔输卵管上皮细胞中的跳跃式细胞器运动不太可能由微管-动力蛋白相互作用引起。

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