Schrøder Rikke Louise, Strøbaek Dorte, Olesen Søren-Peter, Christophersen Palle
NeuroSearch A/S, 93 Pederstrupvej, DK 2750 Ballerup, Denmark.
Pflugers Arch. 2003 Aug;446(5):607-16. doi: 10.1007/s00424-003-1116-x. Epub 2003 Jul 8.
The compound BMS-204352 has been targeted for use against acute ischemic stroke, due to its activation of the large-conductance Ca2+-activated K-channel (BK). We have previously described that the racemate (+/-)BMS-204352 reversibly modulates KCNQ4 voltage dependency. Here we show that (+/-)BMS-204352 also induces a voltage-independent KCNQ4 current. The channels were stably expressed in human embryonic kidney cells (HEK293), and investigated by use of the whole-cell mode of the patch-clamp technique. (+/-)BMS-204352 was applied extracellularly (10 microM) in order to precipitate the robust appearance of the voltage-independent current. The voltage-independent KCNQ4 currents were recorded as instantaneous increases in currents upon hyperpolarizing or depolarizing voltage steps elicited from holding potentials of -90 or -110 mV. The voltage-independent current reversed at the equilibrium potential for potassium ( E(K)), hence was carried by a K+ conductance, and was blocked by the selective KCNQ channel blockers XE991 and linopirdine. Similar results were obtained with KCNQ4 channels transiently transfected into Chinese hamster ovary cells (CHO). When (+/-)BMS-204352 was applied to stably expressed BK channels, only the voltage dependency was modulated. Retigabine, the classic activator of KCNQ channels, did not induce voltage-independent currents. Our data indicate that KCNQ4 channels may conduct voltage-dependent and voltage-independent currents in the presence of (+/-)BMS-204352.
化合物BMS - 204352因能激活大电导钙激活钾通道(BK)而被用于治疗急性缺血性中风。我们之前曾描述过,外消旋体(±)BMS - 204352可可逆地调节KCNQ4的电压依赖性。在此我们表明,(±)BMS - 204352还可诱导一种非电压依赖性的KCNQ4电流。这些通道在人胚肾细胞(HEK293)中稳定表达,并采用膜片钳技术的全细胞模式进行研究。为了促使非电压依赖性电流的强劲出现,将(±)BMS - 204352以细胞外给药的方式(10微摩尔)应用。非电压依赖性KCNQ4电流通过从 - 90或 - 110毫伏的保持电位引发的超极化或去极化电压阶跃时电流的瞬时增加来记录。非电压依赖性电流在钾的平衡电位(E(K))处反转,因此由钾电导携带,并被选择性KCNQ通道阻滞剂XE991和利诺吡啶阻断。在中国仓鼠卵巢细胞(CHO)中瞬时转染KCNQ4通道也获得了类似结果。当将(±)BMS - 204352应用于稳定表达的BK通道时,仅调节了电压依赖性。KCNQ通道的经典激活剂瑞替加滨并未诱导非电压依赖性电流。我们的数据表明,在存在(±)BMS - 204352的情况下,KCNQ4通道可能传导电压依赖性和非电压依赖性电流。