Characterization of the plasma and mitochondrial membrane potentials of alveolar type II cells by the use of ionic probes.

Characterization of the plasma and mitochondrial membrane potentials of alveolar type II cells by the use of ionic probes.
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使用离子探针表征 II 型肺泡细胞的血浆和线粒体膜电位。

DOI:
10.1016/0005-2736(84)90536-4
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发表时间:
1984
期刊:
Biochimica et biophysica acta
影响因子:
--
通讯作者:
Notter,RH
Notter,RH
中科院分区:
--
文献类型:
--
作者:
Gallo,RL;Finkelstein,JN;Notter,RH

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采用亲脂性阳离子三苯基甲基磷(TPMP+)和钾类似物Rb+监测新分离的兔ⅱ型肺泡上皮细胞的膜电位(ΔΨ)。II型细胞在含5 μM四苯基硼的Hanks平衡盐溶液中,在37°C下快速积累TPMP+,但这种积累部分是由于TPMP+与细胞的非膜电位依赖性结合。发现溶血磷脂酰胆碱(lysoPC)消除ΔΨ并允许校正结合的TPMP+或Rb+。结合校正后留在细胞中的TPMP+代表线粒体和质膜电位依赖性积累的总和。II型细胞Rb+的积累与线粒体膜电位无关,其跨质膜Rb+分布电位为- 62.9±4 mV。通过估算线粒体被羰基氰-氯苯腙(CCCP)去极化的II型细胞中TPMP+的质膜电位依赖性积累,得到了类似的值。CCCP处理引起的TPMP+释放也使跨线粒体膜电位估计为- 141.8±10 mV。这些膜电位测量技术被发现对一些抑制剂和离子载体诱导的ΔΨ变化很敏感。测量II型肺细胞膜电位的能力,以及各种药物引起的变化,应该有助于表征这种肺表面活性物质产生细胞的功能反应。
The lipophilic cation triphenylmethylphosphonium (TPMP+) and the potassium analog Rb+, were used to monitor the membrane potential (ΔΨ) of freshly isolated rabbit type II alveolar epithelial cells. Type II cells were found to accumulate TPMP+rapidly at 37°C in Hanks' balanced-salt solution with 5 μM tetraphenyl boron, but this accumulation was partially due to non-membrane potential dependent binding of TPMP+to the cell. Lysophosphatidylcholine (lysoPC) was found to abolish ΔΨ and permitted correction for bound TPMP+or Rb+. TPMP+remaining in the cell following correction for binding represents the sum of mitochondrial and plasma membrane potential dependent accumulation. The accumulation of Rb+by the type II cell was found to be independent of the mitochondrial membrane potential and indicated a trans-plasma membrane Rb+distribution potential of −62.9 ± 4 mV. A similar value was obtained by estimating the plasma membrane potential dependent accumulation of TPMP+in type II cells whose mitochondria were depolarized with carbonylcyanidem-chlorophenylhydrazone (CCCP). The release of TPMP+due to CCCP treatment also permitted an estimation for the trans-mitochondrial membrane potential of −141.8 ± 10 mV. These techniques of membrane potential measurements were found to be sensitive to changes in ΔΨ induced by a number of inhibitors and ionophores. The ability to measure the membrane potential of the type II pneumocyte, and the changes caused by various agents, should be useful in characterizing the functional responses of this pulmonary surfactant producing cell.
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