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血清饥饿和血清刺激的瑞士3T3小鼠成纤维细胞中的微管动力学:对血清诱导的收缩性与微管之间关系的启示

Microtubule dynamics in serum-starved and serum-stimulated Swiss 3T3 mouse fibroblasts: implications for the relationship between serum-induced contractility and microtubules.

作者信息

Danowski B A

机构信息

Department of Biology, Union College, Schenectady, New York 12308, USA.

出版信息

Cell Motil Cytoskeleton. 1998;40(1):1-12. doi: 10.1002/(SICI)1097-0169(1998)40:1<1::AID-CM1>3.0.CO;2-K.

Abstract

It has been established that cell contractility can be stimulated with low or depolymerizing doses of microtubule (MT) poisons. In addition, low doses of nocodazole and vinblastine have recently been shown to decrease MT dynamics in vivo. In this study, investigated whether there is a direct, or reciprocal feedback-type relationship between contractility and microtubule dynamics, by examining MT dynamic behavior in live cells under conditions where contractility is known to be altered. Quiescent, serum-starved Swiss 3T3 mouse fibroblasts have been shown to be weakened in their contractility; serum stimulation increases cell contractility and causes the formation of stress fibers and adhesion plaques. Growing (control), quiescent (Go), and serum-stimulated cells were injected with rhodamine-tubulin, and MT dynamics were determined by analysis of MT length changes obtained from digitized images of the extreme periphery of the cells, where the MT ends were readily apparent. The MTs in quiescent cells were less dynamic than those in control cells: the growth and shortening rates were reduced by 30% and 45%, respectively. Dynamicity decreased by 47%, and the MTs spent more time in pause. After serum stimulation, MT growth rate, dynamicity, and time spent in pause returned to control cell levels. Although the shortening rate increased by 28%, it remained significantly lower than in control cells. In this system, the serum-induced increase in contractility was accompanied by an increase in MT dynamics. However, increased contractility stimulated with low doses of MT poisons is known to be accompanied by a decrease in MT dynamics. These results suggest that the relationship between MT dynamics and contractility is an indirect one.

摘要

已经证实,低剂量或解聚剂量的微管(MT)毒物可刺激细胞收缩性。此外,最近已表明低剂量的诺考达唑和长春花碱可降低体内微管动力学。在本研究中,通过在已知收缩性改变的条件下检查活细胞中的微管动态行为,研究收缩性与微管动力学之间是否存在直接或相互反馈型关系。静止、血清饥饿的瑞士3T3小鼠成纤维细胞已被证明其收缩性减弱;血清刺激可增加细胞收缩性并导致应力纤维和黏附斑的形成。将生长(对照)、静止(G0)和血清刺激的细胞注射罗丹明微管蛋白,并通过分析从细胞最外周的数字化图像获得的微管长度变化来确定微管动力学,在该部位微管末端很容易观察到。静止细胞中的微管动态性低于对照细胞:生长和缩短速率分别降低了30%和45%。动态性降低了47%,微管处于暂停状态的时间更长。血清刺激后,微管生长速率、动态性和暂停时间恢复到对照细胞水平。虽然缩短速率增加了28%,但仍显著低于对照细胞。在该系统中,血清诱导的收缩性增加伴随着微管动力学的增加。然而,已知低剂量微管毒物刺激导致的收缩性增加伴随着微管动力学的降低。这些结果表明,微管动力学与收缩性之间的关系是间接的。

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