Dupré-Aucouturier S, Penhoat A, Rougier O, Bilbaut A
Université Claude Bernard Lyon I, UMR-CNRS 5123, Physiologie des Eléments Excitables, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne, France.
J Membr Biol. 2004 May 15;199(2):99-111. doi: 10.1007/s00232-004-0680-0.
In a previous study performed on zona fasciculata (ZF) cells isolated from calf adrenal glands, we identified an ACTH-induced Cl- current involved in cell membrane depolarization. In the present work, we describe a volume-sensitive Cl- current and compare it with the ACTH-activated Cl- current. Experiments were performed using the whole-cell patch-clamp recording method, video microscopy and cortisol-secretion measurements. In current-clamp experiments, hypotonic solutions induced a membrane depolarization to -22 mV. This depolarization, correlated with an increase in the membrane conductance, was sensitive to different Cl- channel inhibitors. In voltage-clamp experiments, hypotonic solution induced a membrane current that slowly decayed and reversed at -21 mV. This ionic current displayed no time dependence and showed a slight outward rectification. It was blocked to variable extent by different conventional Cl- channel inhibitors. Under hypotonic conditions, membrane depolarizations were preceded by an increase in cell volume that was not detected under ACTH stimulation. It was concluded that hypotonic solution induced cell swelling, which activated a Cl- current involved in membrane depolarization. Although cell volume change was not observed in the presence of ACTH, biophysical properties and pharmacological profile of the volume-sensitive Cl- current present obvious similarities with the ACTH-activated Cl- current. As compared to ACTH, hypotonic solutions failed to trigger cortisol production that was weakly stimulated in the presence of high-K+ solution. This shows that in ZF cells, membrane depolarization is not a sufficient condition to fully activate secretory activities.
在之前一项对从小牛肾上腺分离出的束状带(ZF)细胞进行的研究中,我们鉴定出一种促肾上腺皮质激素(ACTH)诱导的氯离子电流,它参与细胞膜去极化过程。在本研究中,我们描述了一种容积敏感性氯离子电流,并将其与ACTH激活的氯离子电流进行比较。实验采用全细胞膜片钳记录法、视频显微镜和皮质醇分泌测量技术进行。在电流钳实验中,低渗溶液诱导细胞膜去极化至-22 mV。这种去极化与膜电导增加相关,对不同的氯离子通道抑制剂敏感。在电压钳实验中,低渗溶液诱导出一种缓慢衰减且在-21 mV时反转的膜电流。这种离子电流不显示时间依赖性,表现出轻微的外向整流特性。它被不同的传统氯离子通道抑制剂不同程度地阻断。在低渗条件下,细胞膜去极化之前细胞体积会增加,而在ACTH刺激下未检测到这种情况。得出的结论是,低渗溶液诱导细胞肿胀,进而激活一种参与细胞膜去极化的氯离子电流。尽管在ACTH存在时未观察到细胞体积变化,但容积敏感性氯离子电流的生物物理特性和药理学特征与ACTH激活的氯离子电流存在明显相似性。与ACTH相比,低渗溶液未能触发皮质醇分泌,而在高钾溶液存在时皮质醇分泌受到微弱刺激。这表明在ZF细胞中,细胞膜去极化并非充分激活分泌活动的条件。