Xiong Z, Sperelakis N, Noffsinger A, Fenoglio-Preiser C
Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Ohio 45267, USA.
Am J Physiol. 1995 Sep;269(3 Pt 1):G378-85. doi: 10.1152/ajpgi.1995.269.3.G378.
Voltage-gated Ca2+ currents were investigated in single smooth muscle cells freshly isolated from the circular layer of the human colon (ascending and descending portions) using the whole cell voltage-clamp technique. Tissue samples were obtained at the time of therapeutic surgery. In physiological salt solution (containing 2 mM Ca2+), an inward current was observed when the cell membrane was depolarized in the presence of tetrodotoxin. This current disappeared when Ca2+ was removed from the bath solution and was inhibited when Ca2+ channel blockers were applied, indicating that the inward current was a Ca2+ current (ICa). Changing the holding potential (HP) from -100 mV to more positive potentials (e.g., -60 and -40 mV) markedly decreased the amplitude of ICa. The voltage dependence of steady-state activation and inactivation was represented by Boltzmann distributions; there was a substantial amount of overlap (window current) between -60 and -10 mV. A fast-inactivating ICa component followed by a slow-inactivating ICa component was observed in some cells from both ascending and descending colons. The fast ICa component was observed only when cells were held at -80 or -100 mV, and had a more negative threshold potential (-70 to -60 mV). This component was sensitive to low concentrations of Ni2+ (30 microM) but was resistant to nifedipine (10-20 microM). In contrast, the slow (sustained) ICa component was observed at all HPs (-40 to -100 mV) and had a more positive threshold potential (about -40 mV). This component was insensitive to low concentration of Ni2+ but was sensitive to nifedipine and BAY K 8644.(ABSTRACT TRUNCATED AT 250 WORDS)
采用全细胞电压钳技术,对从人结肠环形肌层(升结肠和降结肠部分)新鲜分离的单个平滑肌细胞中的电压门控钙电流进行了研究。组织样本在治疗性手术时获取。在生理盐溶液(含2 mM钙离子)中,当细胞膜在河豚毒素存在下发生去极化时,可观察到内向电流。当从浴液中去除钙离子时,该电流消失,而应用钙离子通道阻滞剂时则受到抑制,这表明内向电流是钙电流(ICa)。将钳制电位(HP)从 -100 mV改变为更正的电位(如 -60和 -40 mV)会显著降低ICa的幅度。稳态激活和失活的电压依赖性由玻尔兹曼分布表示;在 -60和 -10 mV之间存在大量重叠(窗电流)。在升结肠和降结肠的一些细胞中观察到一个快速失活的ICa成分,随后是一个缓慢失活的ICa成分。快速ICa成分仅在细胞保持在 -80或 -100 mV时观察到,且具有更负的阈电位(-70至 -60 mV)。该成分对低浓度的镍离子(30 microM)敏感,但对硝苯地平(10 - 20 microM)有抗性。相比之下,缓慢(持续)的ICa成分在所有钳制电位(-40至 -100 mV)下均能观察到,且具有更正的阈电位(约 -40 mV)。该成分对低浓度的镍离子不敏感,但对硝苯地平和BAY K 8644敏感。(摘要截于250字)