High Cell Density Upregulates Calcium Oscillation by Increasing Calcium Store Content via Basal Mitogen-Activated Protein Kinase Activity.
High Cell Density Upregulates Calcium Oscillation by Increasing Calcium Store Content via Basal Mitogen-Activated Protein Kinase Activity.
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DOI：
10.1371/journal.pone.0137610
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kudo Y
中科院分区：
文献类型：
--
作者：
Morita M;Nakane A;Fujii Y;Maekawa S;Kudo Y
非兴奋性细胞的钙释放通常是振荡和非振荡模式的组合，影响钙动力学的因素仍有待确定。在这里，我们报告的影响，细胞密度的钙增加模式的克隆细胞系。大多数接种在1.5 × 104/cm 2的HeLa细胞显示出响应组胺和ATP的钙振荡，而接种在0.5 × 104/cm 2的细胞主要显示出短暂和持续的钙增加。细胞密度也以类似的方式影响HEK 293细胞对ATP的反应。高细胞密度增加了促分裂原活化蛋白（MAP）激酶的基础活性和钙库含量，MAP激酶抑制剂U 0126下调了钙振荡和钙库含量。因此，MAP激酶介导的钙储存调节可能是细胞密度对钙振荡影响的基础。HeLa细胞的钙增加模式是保守的，在任何组胺浓度测试，而组胺H1受体的过度表达，这有力地增加组胺诱导的肌醇磷脂水解，转换钙振荡持续钙增加只有在高组胺浓度。因此，调节肌醇磷脂代谢的结果与改变细胞密度的结果不同，表明细胞密度的影响不归因于肌醇磷脂代谢。总的来说，我们的研究结果表明，钙增加模式的非兴奋性细胞反映钙储存，这是调节的基础MAP激酶活性的影响下的细胞密度。
Calcium releases of non-excitable cells are generally a combination of oscillatory and non-oscillatory patterns, and factors affecting the calcium dynamics are still to be determined. Here we report the influence of cell density on calcium increase patterns of clonal cell lines. The majority of HeLa cells seeded at 1.5 x 104/cm2 showed calcium oscillations in response to histamine and ATP, whereas cells seeded at 0.5 x 104/cm2 largely showed transient and sustained calcium increases. Cell density also affected the response of HEK293 cells to ATP in a similar manner. High cell density increased the basal activity of the mitogen-activated protein (MAP) kinase and calcium store content, and both calcium oscillation and calcium store content were down-regulated by a MAP kinase inhibitor, U0126. Thus, MAP kinase-mediated regulation of calcium store likely underlie the effect of cell density on calcium oscillation. Calcium increase patterns of HeLa cells were conserved at any histamine concentrations tested, whereas the overexpression of histamine H1 receptor, which robustly increased histamine-induced inositol phospholipid hydrolysis, converted calcium oscillations to sustained calcium increases only at high histamine concentrations. Thus, the consequence of modulating inositol phospholipid metabolism was distinct from that of changing cell density, suggesting the effect of cell density is not attributed to inositol phospholipid metabolism. Collectively, our results propose that calcium increase patterns of non-excitable cells reflect calcium store, which is regulated by the basal MAP kinase activity under the influence of cell density.