Manipulation of cell volume and membrane pore comparison following single cell permeabilization with 60- and 600-ns electric pulses.
Manipulation of cell volume and membrane pore comparison following single cell permeabilization with 60- and 600-ns electric pulses.
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DOI：
10.1016/j.bbamem.2010.12.012
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发表时间：
2011-03
期刊：
影响因子：
--
通讯作者：
Pakhomov, Andrei G
中科院分区：
文献类型：
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作者：
Nesin, Olena M;Pakhomova, Olga N;Xiao, Shu;Pakhomov, Andrei G
强烈的纳秒持续时间的电脉冲（nsEP）打开细胞质膜中稳定的纳米孔，随后由于水的吸收或排出而引起细胞体积的变化，这是由细胞内外的孔不可渗透溶质的渗透压平衡调节的。用聚乙二醇和糖等渗置换NaCl溶液，测定了50，60 ns EP（10 Hz，~13 kV/cm）和5，600 ns EP（1 Hz，~6 kV/cm）对GH 3细胞开放的孔的大小。这种替换减少了细胞肿胀和/或将其转变为短暂或持续的收缩，这取决于测试溶质可渗透的孔的可用性。出乎意料的是，溶质置换显示，对于由60-和600-ns处理打开的孔的相同积分面积（如细胞体积变化所示），孔径是相似的。然而，600纳秒曝光触发显着更高的细胞摄取的丙啶。我们的结论是，600纳秒EP打开了更大数量的更大的（可渗透丙铵的孔），但在整个孔群中的较大孔的分数是不够的，有助于细胞体积的变化。对于60-和600-ns的曝光，细胞体积的变化是由直径小于0.9 nm的孔确定的，然而，直径随着nsEP强度的增加而增加。
Intense nanosecond-duration electric pulses (nsEP) open stable nanopores in cell plasma membrane, followed by cell volume changesdue to water uptake or expulsion, as regulated by the osmolality balance of pore-impermeable solutes inside and outside the cell. The size of pores opened by 50, 60-ns EP (10 Hz, ~13 kV/cm) and 5, 600-ns EP (1 Hz, ~6 kV/cm) in GH3 cells was estimated by isoosmotic replacement of bath NaCl with (polyethylene glycols and sugars. Such replacement reduced cell swelling and/or turned it into a transient or sustained shrinking, depending on the availability of pores permeable to the test solute. Unexpectedly, solute substitutions showed that for the same integral area of pores opened by 60- and 600-ns treatments (as indicated by cell volume changes), the pore sizes were similar. However, the 600-ns exposure triggered significantly higher cell uptake of propidium. We concluded that 600-ns EP opened a greater number of larger (propidium-permeable pores), but the fraction of the larger pores in the entire pore population was insufficient to contribute to cell volume changes. For both the 60- and 600-ns exposures, cell volume changes were determined by pores smaller than 0.9 nm in diameter; however, the diameter increased with increasing the nsEP intensity.