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Microtubule Disruption with BAPTA and Dimethyl BAPTA by a Calcium Chelation-lndependent Mechanism in 3T3-L1 Adipocytes

机译:BAPTA和二甲基BAPTA通过钙螯合独立机制在3T3-L1脂肪细胞中的微管破坏。

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摘要

While the physiological role for calcium in the insulin action on glucose transport has been disputed, it was reassessed in a recent study by using a calcum chelator, l,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, tetra(acetoxymethyl) ester (BAPTA-AM). Although BAPTA has been widely used to study the role for calcium in a variety of cell functions, it has also been suggested to have properties unrelated to the calcium chelating activity. Here, we investigated the effects of BAPTA and dimethyl BAPTA on the cytoskeletons in 3T3-L1 adipocytes. Both calcium chelators were successfully loaded in 3T3-L1 adipocytes and inhibited endothelin-1-induced cytosolic calcium elevation. Confocal fluorescence microscopy revealed that BAPTA and dimethyl BAPTA caused profound depolymerization of the microtubules without affecting the cortical actin filaments in 3T3-L1 adipocytes. Biochemical quantification also showed that BAPTA and dimethyl BAPTA significantly decreased the amount of polymerized tubulin but had little effect on filamentous actin. Consistent with these results, GLUT4-positive perinuclear compartments were dispersed throughout the cytoplasm in BAPTA- or dimethyl BAPTA-loaded adipocytes. Intriguingly, these calcium chelators did not disrupt the microtubules in undifferentiated preadipocytes. The microtubule-depolymerizing property of BAPTA and dimethyl BAPTA is unrelated to calcium chelation, since the microtubules were resistant to depletion of cytosolic calcium by using a calcium ionophore A23187. Insulin-stimulated glucose transport was not affected by cytosolic calcium depletion with A23187, but significantly inhibited with BAPTA and dimethyl BAPTA to the extent similar to that with nocodazole. BAPTA and its derivatives should be used with caution in studies of cytoskeleton-related cell functions.
机译:尽管人们对钙在胰岛素对葡萄糖转运的作用中的生理作用存在争议,但在最近的一项研究中,它通过使用钙钙螯合剂l,2-双(邻氨基苯氧基)乙烷-N,N,N',N进行了重新评估。 -四乙酸,四(乙酰氧基甲基)酯(BAPTA-AM)。尽管BAPTA已被广泛用于研究钙在多种细胞功能中的作用,但也有人建议其具有与钙螯合活性无关的性质。在这里,我们研究了BAPTA和二甲基BAPTA对3T3-L1脂肪细胞中细胞骨架的影响。两种钙螯合剂均成功加载到3T3-L1脂肪细胞中,并抑制了内皮素1诱导的胞质钙升高。共聚焦荧光显微镜显示,BAPTA和二甲基BAPTA引起微管的深度解聚,而不影响3T3-L1脂肪细胞中的皮质肌动蛋白丝。生化定量分析还表明,BAPTA和二甲基BAPTA显着降低了聚合微管蛋白的量,但对丝状肌动蛋白的影响很小。与这些结果一致,GLUT4阳性核周区室分散在BAPTA或二甲基BAPTA加载的脂肪细胞中的整个细胞质中。有趣的是,这些钙螯合剂并未破坏未分化的前脂肪细胞中的微管。 BAPTA和二甲基BAPTA的微管解聚特性与钙螯合无关,因为微管通过使用钙离子载体A23187可以抵抗细胞质钙的消耗。胰岛素刺激的葡萄糖转运不受A23187的胞质钙耗竭的影响,但被BAPTA和二甲基BAPTA显着抑制,其程度与诺考达唑相似。在研究细胞骨架相关的细胞功能时,应谨慎使用BAPTA及其衍生物。

著录项

  • 来源
    《Endocrine journal》 |2009年第2期|235-243|共9页
  • 作者单位

    Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan;

    Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan;

    Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan;

    Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan;

    Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan;

    Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-machi, Maebashi 371-8512, Japan;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    insulin; calcium; glucose transporter; BAPTA; microtubule;

    机译:胰岛素;钙;葡萄糖转运蛋白BAPTA;微管;
  • 入库时间 2022-08-18 01:33:43

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