Duan D, Nattel S
Department of Medicine, Montreal Heart Institute, Quebec, Canada.
Circ Res. 1994 Oct;75(4):789-95. doi: 10.1161/01.res.75.4.789.
A variety of potentially important macroscopic Cl- currents have been described in the heart. Although the single-channel properties of the cAMP-dependent current (ICl.cAMP) have been well described, the single-channel equivalents of the other forms of cardiac Cl- current remain unknown. Unlike ICl.cAMP, many of these currents show prominent outward rectification in the presence of symmetrical transmembrane Cl- gradients and sensitivity to disulfonic stilbene Cl- transport blockers. We used the patch-clamp technique to search for single Cl- channels in inside-out patches from rabbit atrial cell membranes, under conditions minimizing the chances of observing channels carrying Na+, Ca2+, or K+. Under symmetrical Cl- conditions, single-channel activity was seen in 14 (9%) of 155 patches. Channels showed strong outward rectification and a unitary conductance of 60 +/- 3 picosiemens (mean +/- SEM) at positive voltages. The current-voltage relation was not altered by replacement of cations by the impermeant cation N'-methyl-D-glucamine (NMDG) and shifted as expected for a Cl(-)-selective channel when methanesulfonate was substituted for Cl-. The Cl- transport blockers DIDS (diisothiocyanatostilbene-2,2'-disulfonic acid, 100 mumol/L) and SITS (4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid, 1 mmol/L) strongly and reversibly inhibited channel activity when added to the bath and caused channel flickering suggesting open-channel block. Ensemble-average currents showed no time dependence, and the form of the ensemble-average current-voltage relation was similar to that of macroscopic background Cl- current.(ABSTRACT TRUNCATED AT 250 WORDS)
心脏中已描述了多种可能具有重要意义的宏观氯离子电流。尽管对环磷酸腺苷依赖性电流(ICl.cAMP)的单通道特性已有充分描述,但心脏中其他形式氯离子电流的单通道等效物仍不清楚。与ICl.cAMP不同,这些电流中的许多在跨膜氯离子梯度对称的情况下表现出明显的外向整流特性,并且对二磺酸芪类氯离子转运阻滞剂敏感。我们使用膜片钳技术,在尽量减少观察到携带钠离子、钙离子或钾离子通道可能性的条件下,从兔心房细胞膜的内向外膜片中寻找单个氯离子通道。在氯离子对称条件下,155个膜片中的14个(9%)观察到了单通道活性。通道在正电压下表现出强烈的外向整流特性,单位电导为60±3皮西门子(平均值±标准误)。用非渗透性阳离子N'-甲基-D-葡萄糖胺(NMDG)替代阳离子时,电流-电压关系未改变,而用甲磺酸盐替代氯离子时,电流-电压关系如氯离子选择性通道预期的那样发生了偏移。当向浴槽中加入氯离子转运阻滞剂二异硫氰酸芪-2,2'-二磺酸(DIDS,100 μmol/L)和4-乙酰氨基-4'-异硫氰酸芪-2,2'-二磺酸(SITS,1 mmol/L)时,它们强烈且可逆地抑制通道活性,并导致通道闪烁,提示存在开放通道阻断。整体平均电流无时间依赖性,整体平均电流-电压关系的形式与宏观背景氯离子电流相似。(摘要截短于250字)