低水平钙对哺乳动物细胞膜电通透化的调控

Control by Low Levels of Calcium of Mammalian Cell Membrane Electropermeabilization.

作者信息

Ciobanu Florin, Golzio Muriel, Kovacs Eugenia, Teissié Justin

机构信息

Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, Toulouse, France.

University Carol Davila, Bucarest, Romania.

出版信息

J Membr Biol. 2018 Apr;251(2):221-228. doi: 10.1007/s00232-017-9981-y. Epub 2017 Aug 20.

Abstract

Electric pulses, when applied to a cell suspension, induce a reversible permeabilization of the plasma membrane. This permeabilized state is a long-lived process (minutes). The biophysical molecular mechanisms supporting the membrane reorganization associated to its permeabilization remain poorly understood. Modeling the transmembrane structures as toroidal lipidic pores cannot explain why they are long-lived and why their resealing is under the control of the ATP level. Our results describe the effect of the level of free Calcium ions. Permeabilization induces a Ca burst as previously shown by imaging of cells loaded with Fluo-3. But this sharp increase is reversible even when Calcium is present at a millimolar concentration. Viability is preserved to a larger extent when submillimolar concentrations are used. The effect of calcium ions is occurring during the resealing step not during the creation of permeabilization as the same effect is observed if Ca is added in the few seconds following the pulses. The resealing time is faster when Ca is present in a dose-dependent manner. Mg is observed to play a competitive role. These observations suggest that Ca is acting not on the external leaflet of the plasma membrane but due to its increased concentration in the cytoplasm. Exocytosis will be enhanced by this Ca burst (but hindered by Mg) and occurs in the electropermeabilized part of the cell surface. This description is supported by previous theoretical and experimental results. The associated fusion of vesicles will be the support of resealing.

摘要

当将电脉冲施加到细胞悬液上时,会诱导质膜发生可逆的通透性改变。这种通透状态是一个持续较长时间的过程(数分钟)。支持与其通透性相关的膜重组的生物物理分子机制仍知之甚少。将跨膜结构建模为环形脂质孔无法解释它们为何寿命长以及它们的重新封闭为何受ATP水平的控制。我们的结果描述了游离钙离子水平的影响。如先前用Fluo-3负载的细胞成像所示,通透性改变会引发钙爆发。但即使钙离子浓度为毫摩尔级,这种急剧增加也是可逆的。使用亚毫摩尔浓度时,细胞活力能在更大程度上得以保留。钙离子的作用发生在重新封闭步骤期间,而不是在通透性形成过程中,因为如果在脉冲后的几秒钟内添加钙,会观察到相同的效果。当钙以剂量依赖性方式存在时,重新封闭时间会更快。观察到镁起到竞争作用。这些观察结果表明,钙的作用不是作用于质膜的外部小叶,而是由于其在细胞质中浓度的增加。这种钙爆发会增强胞吐作用(但会受到镁的阻碍),并发生在细胞表面的电通透部分。这一描述得到了先前理论和实验结果的支持。相关的囊泡融合将是重新封闭的支撑。

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