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培养的神经和肌肉细胞中补体介导的膜损伤的电学测量。

Electrical measurements of complement-mediated membrane damage in cultured nerve and muscle cells.

作者信息

Stephens C L, Henkart P A

出版信息

J Immunol. 1979 Feb;122(2):455-8.

PMID:762431
Abstract

Cultured neuroblastoma-glioma hybrid cells and primary mouse and rat muscle cells were studied by using intracellular microelectrodes to monitor membrane electrical potential and resistance changes during complement-mediated lysis. The cell membrane was TNP modified under mild conditions and subsequently coated with rabbit IgG anti-TNP, with no electrical changes observed. However, upon addition of guinea pig C the membrane potential dropped from approximately -50 mv to less than -12 mv within a few minutes, with parallel decreases in electrical resistance. Ten to 60 min after these electrical changes the cells became stainable with trypan blue. No electrical changes or trypan blue staining was observed in the absence of antibody or with heat-inactivated C. With more dilute C so that only a fraction of cells became trypan blue positive, all cells nevertheless showed the membrane electrical changes; surviving cells recovered their original membrane properties within 1 hr. Thus the C-mediated damage to the membrane measured electrically is not in itself sufficient to ensure the subsequent death of the cell. The early electrical changes observed appear to be comparable to increases in 86Rb efflux measured by others.

摘要

通过使用细胞内微电极监测补体介导的细胞裂解过程中膜电位和电阻的变化,对培养的神经母细胞瘤 - 胶质瘤杂交细胞以及原代小鼠和大鼠肌肉细胞进行了研究。在温和条件下对细胞膜进行三硝基苯(TNP)修饰,随后用兔抗TNP IgG包被,未观察到电变化。然而,加入豚鼠补体C后,膜电位在几分钟内从约 -50 mV降至低于 -12 mV,同时电阻也相应降低。这些电变化发生后10至60分钟,细胞能够被台盼蓝染色。在没有抗体或使用热灭活补体C的情况下,未观察到电变化或台盼蓝染色。使用更稀释的补体C,使得只有一部分细胞变为台盼蓝阳性,但所有细胞均显示出膜电变化;存活的细胞在1小时内恢复其原始膜特性。因此,通过电测量的补体介导的膜损伤本身不足以确保细胞随后的死亡。观察到的早期电变化似乎与其他人测量的86Rb外流增加相当。

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