Effects of exogenous ganglioside GM1 on different stages of cell spreading studied by directly quantifying spreading rate

Effects of exogenous ganglioside GM1 on different stages of cell spreading studied by directly quantifying spreading rate
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通过直接量化扩散速率研究外源神经节苷脂 GM1 对细胞扩散不同阶段的影响

DOI:
10.3109/15419061.2012.749245
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发表时间:
2012-10
影响因子:
--
通讯作者:
Yong Chen
Yong Chen
中科院分区:
生物4区
文献类型:
--
作者:
Jie Huang;Wenxiang Shao;Li Wu;Wen Yang;Yong Chen

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摘要我们开发了新的方法来直接量化细胞扩散率。通过与传统方法的比较,我们发现胎牛血清(FCS)的促进作用或外源性神经节苷脂GM 1的抑制作用发生在细胞铺展的不同阶段。GM 1主要影响HUVECs细胞伸展的早期和晚期。在0.5%FCS的存在下,GM 1显著损害了基于区域的铺展率(127.4 ± 35.7 μm2/h和22.2 ± 3.8 μm2/h)与对照组相比,细胞铺展的0-0.5 h和晚期(12-24 h)阶段(分别为238.1 ± 11.7 μm2/h和35.4 ± 19.5 μm2/h),这通过在细胞铺展期间GM 1诱导的肌动蛋白丝平均长度的变化的数据得到证实。对无GM 1或GM 1处理的HUVECs冷诱导的去铺展的实时观察和定量进一步证实了GM 1可以影响细胞去铺展过程,在不同阶段具有抑制(0-10 min)或增强(10-20 min或40-50 min)作用。该方法可用于研究其他试剂对细胞铺展不同阶段的影响。
Abstract We developed novel methods to directly quantify cell spreading rate. By comparing our methods with traditional methods, we found that the enhancement effects of fetal calf serum (FCS) or the inhibitory effects of exogenous ganglioside GM1 occurred at different stages of cell spreading. GM1 mainly influenced the early and late stages of cell spreading of HUVECs. In the presence of 0.5% FCS, GM1 significantly impaired the area-based spreading rates (127.4 ± 35.7 μm2/h and 22.2 ± 3.8 μm2/h, respectively) on the early (0–0.5 h) and late (12–24 h) stages of cell spreading compared with the controls (238.1 ± 11.7 μm2/h and 35.4 ± 19.5 μm2/h, respectively), which was confirmed by the data on the GM1-induced changes in average length of actin filaments during cell spreading. The real-time observation and quantification of cold-induced de-spreading of GM1-free or GM1-treated HUVECs further confirmed that GM1 can influence cell de-spreading process having inhibitory (0–10 min) or enhancement (10–20 min or 40–50 min) effects on different stages. The methods can be recruited for investigating effects of other reagents on different stages of cell spreading.
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