胚胎心肌细胞原代培养过程中原肌球蛋白的变化

M.A. Muros, A.E. Aránega, C. Vélez, F.J. González, J.E. Fernández, L. Alvarez, A. Aránega
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引用次数: 2

摘要

本实验选用汉堡和汉密尔顿29期(HH 29)鸡心肌细胞,观察培养14 d后原肌球蛋白在鸡心肌细胞中的表达情况。在14天的细胞培养过程中,细胞形态的变化伴随着原肌球蛋白在不同细胞区室的重新分布。我们使用FACScan, SDS-PAGE和密度分析来定量细胞原肌球蛋白总量和该蛋白在不同细胞组分中的浓度。原肌球蛋白主要存在于细胞骨架部分,而不在细胞质中。当我们比较不同发育阶段细胞的SDS-PAGE密度值时,我们发现在HH 19中,原肌球蛋白在细胞质中比在细胞骨架部分中更丰富。到HH 29时,这两个部分已经倒置,在HH 39时,原肌球蛋白在细胞骨架中的含量明显高于细胞质部分。在IFI分析中,发现原肌球蛋白根据表达该蛋白的细胞区域以不同的模式标记应力纤维样结构(SFL)。
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Changes in tropomyosin during primary culture of embryonic myocardiocytes

We chose the Hamburger and Hamilton's stage 29 (HH 29) to investigate the expression of tropomyosin in chick myocardiocytes during 14 days on culture. Throughout 14 days of cell culture, changes in cell morphology were accompained by a redistribution of tropomyosin in different cell compartments. We used FACScan, SDS-PAGE and densitometric analysis to quantify total cell tropomyosin and concentrations of this protein in different cell fractions. Tropomyosin was found mostly in the cytoskeletal fraction than in the cytoplasmic.

When we compared the densitometric values from SDS-PAGE of cells in different stages of development we found that in HH 19, tropomyosin was more abundant in the cytoplasmic than in the cytoskeletal fraction. By HH 29, the two fractions had become inverted, and in HH 39, tropomyosin was clearly more abundant in the cytoskeletal than in the cytoplasmic fraction.

In the IFI analysis, tropomyosin was found to label the Stress fiberlike structures (SFL) in different patterns depending on the area of the cell which expressed this protein.

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