Centro Universitario de Investigaciones Biomédicas, Universidad de Colima, Av. 25 de Julio 965, Colonia Villa San Sebastián, C.P. 28045, Colima, Col., Mexico.
Pflugers Arch. 2014 May;466(5):915-24. doi: 10.1007/s00424-013-1350-9. Epub 2013 Sep 17.
The human intestinal pathogen Giardia lamblia is a flagellated unicellular protozoan with pronounced medical and biological relevance. However, the basic physiology of Giardia trophozoites has been sparsely studied, especially the electrical and ionic properties of their cellular membrane which are virtually unknown. In this study, we were able to record and characterize the macroscopic ionic currents of Giardia trophozoite membrane by electrophysiological methods of the patch clamp technique. Giardia trophozoites showed a high current density (∼600 pA/pF at -140 mV) that was activated upon hyperpolarization. This current was carried by a chloride-selective channel (I Cl-G) and it was the most important determinant of the membrane potential in Giardia trophozoites. Moreover, this conductance was able to carry other halide anions and the sequence of permeability was Br(-) > Cl(-) ≈ I(-) ≫ F(-). Besides the voltage-dependent inward-rectifying nature of I Cl-G, its activation and deactivation kinetics were comparable to those observed in ClC-2 channels. Extracellular pH modified the voltage-dependent properties of I Cl-G, shifting the activation curve from a V 1/2 = -79 ± 1 mV (pH 7.4) to -93 ± 2 mV (pH 8.4) and -112 ± 2 mV (pH 5.4). Furthermore, the maximal amplitude of I Cl-G measured at -100 mV showed dependence to external pH in a bell-shaped fashion reported only for ClC-2 channels. Therefore, our results suggest that I Cl-G possesses several functional properties similar to the mammalian ClC-2 channels.
人类肠道病原体蓝氏贾第鞭毛虫是一种具有明显医学和生物学意义的鞭毛单细胞原生动物。然而,贾第虫滋养体的基本生理学研究甚少,特别是其细胞膜的电学和离子特性几乎未知。在这项研究中,我们能够通过膜片钳技术的电生理方法记录和描述贾第虫滋养体细胞膜的宏观离子电流。贾第虫滋养体表现出高电流密度(在-140 mV 时约为 600 pA/pF),这种电流在超极化时被激活。这种电流由氯离子选择性通道(I Cl-G)携带,是贾第虫滋养体膜电位的最重要决定因素。此外,这种电导能够携带其他卤化物阴离子,并且通透性顺序为 Br(-) > Cl(-) ≈ I(-) ≫ F(-)。除了 I Cl-G 的电压依赖性内向整流性质外,其激活和失活动力学与 ClC-2 通道观察到的相似。细胞外 pH 改变了 I Cl-G 的电压依赖性特性,将激活曲线从 V 1/2 = -79 ± 1 mV(pH 7.4)移至-93 ± 2 mV(pH 8.4)和-112 ± 2 mV(pH 5.4)。此外,在-100 mV 测量的 I Cl-G 的最大幅度表现出对外界 pH 的钟形依赖性,这种依赖性仅在 ClC-2 通道中报道过。因此,我们的结果表明 I Cl-G 具有与哺乳动物 ClC-2 通道相似的几种功能特性。