Winpenny J P, Mathews C J, Verdon B, Wardle C J, Chambers J A, Harris A, Argent B E, Gray M A
Department of Physiological Sciences, University Medical School, Framlington Place, Newcastle upon Tyne NE2 4HH, UK.
Pflugers Arch. 1996 Aug;432(4):644-54. doi: 10.1007/s004240050181.
Using the patch-clamp technique, we have identified a large, outwardly rectifying, Cl--selective whole-cell current in primary cultures of human vas deferens epithelial cells. Whole-cell currents were time- and voltage-dependent and displayed inactivation following depolarising pulses >/= 60 mV. Currents were equally permeable to bromide (PBr/PCl = 1.05 +/- 0.04), iodide (PI/PCl = 1. 06 +/- 0.07) and Cl-, but significantly less permeable to gluconate (PGluc /PCl = 0.23 +/- 0.03). Currents spontaneously increased with time after establishing a whole-cell recording, but could be inhibited by exposure to a hypertonic bath solution which reduced inward currents by 68 +/- 4%. Subsequent exposure of the cells to a hypotonic bath solution led to a 418 +/- 110% increase in inward current, indicating that these currents are regulated by osmolarity. 4,4'-Diisothiocyanatostilbene-2,2'-disulphonic acid (100 microM) produced a rapid and reversible voltage-dependent block (60 +/- 5% and 10 +/- 7% inhibition of current, measured at +/- 60 mV, respectively). Dideoxyforskolin (50 microM) also reduced the volume-sensitive Cl- current, but with a much slower time course, by 41 +/- 13% and 32 +/- 16% (measured at +/- 60 mV, respectively). Tamoxifen (10 microM) had no effect on the whole-cell Cl- current. These results suggest that vas deferens epithelial cells possess a volume-sensitive Cl- conductance which has biophysical and pharmacological properties broadly similar to volume-sensitive Cl- currents previously described in a variety of cell types.
运用膜片钳技术,我们在人输精管上皮细胞原代培养物中鉴定出一种大的、外向整流的、Cl⁻选择性全细胞电流。全细胞电流具有时间和电压依赖性,在去极化脉冲≥60 mV后表现出失活。电流对溴化物(PBr/PCl = 1.05 ± 0.04)、碘化物(PI/PCl = 1.06 ± 0.07)和Cl⁻的通透性相同,但对葡萄糖酸盐的通透性显著较低(PGluc /PCl = 0.23 ± 0.03)。在建立全细胞记录后,电流会随时间自发增加,但可被置于高渗浴液中所抑制,高渗浴液使内向电流降低了68 ± 4%。随后将细胞置于低渗浴液中会导致内向电流增加418 ± 110%,表明这些电流受渗透压调节。4,4'-二异硫氰酸芪-2,2'-二磺酸(100 μM)产生快速且可逆的电压依赖性阻断(在±60 mV测量时,分别抑制电流60 ± 5%和10 ± 7%)。双脱氧佛司可林(50 μM)也降低了容积敏感性Cl⁻电流,但时间进程要慢得多,分别为41 ± 13%和32 ± 16%(在±60 mV测量)。他莫昔芬(10 μM)对全细胞Cl⁻电流无影响。这些结果表明,输精管上皮细胞具有一种容积敏感性Cl⁻电导,其生物物理和药理学特性与先前在多种细胞类型中描述的容积敏感性Cl⁻电流大致相似。