Ponce Arturo, Jimenez-Cardoso Enedina, Eligio-Garcia Leticia
Department of Physiology, Biophysics and Neurosciences, Center for Research and Advanced Studies IPN, México City, Mexico.
Parasitology Research Laboratory, Children Hospital of México "Federico Gomez", Mexico City, México.
Physiol Rep. 2013 Dec 29;1(7):e00186. doi: 10.1002/phy2.186. eCollection 2013 Dec 1.
Despite its importance as a health problem issue, almost nothing is known about the membrane physiology of Giardia lamblia and practically there exist no information so far regarding the variety and properties of ion channels that this protozoan parasite possesses. To address this subject we resorted to an indirect method, consisting in the injection of mRNA and further characterization of ion currents in Xenopus oocytes. In this work, we show that oocytes injected with mRNA isolated from cultured trophozoites of G. lamblia, strain Portland-1 express novel potassium currents that appear over the second day after injection and show time- and voltage-dependent activation followed by a slow inactivation. They start activating at -90 mV, with V1/2 of -30 mV; its time constant of activation (at +60 mV) is 0.11 sec, whereas that of inactivation is 1.92 sec, V1/2 = -44.6 mV. Such K currents were effectively blocked by K channel blockers TEA and 4AP, as well as Ba(2+), quinine, quinidine, charybdotoxin, dendrotoxin-1, capsaicin, margatoxin, and diltiazem. These results suggest that such currents are the result of expression of Giardia's voltage-gated K channels heterologously expressed in Xenopus laevis oocytes.
尽管贾第虫作为一个健康问题很重要,但人们对其膜生理学几乎一无所知,实际上到目前为止也没有关于这种原生动物寄生虫所拥有的离子通道种类和特性的信息。为了解决这个问题,我们采用了一种间接方法,即注射信使核糖核酸(mRNA)并进一步表征非洲爪蟾卵母细胞中的离子电流。在这项工作中,我们表明,注射从波特兰-1株蓝氏贾第鞭毛虫培养的滋养体中分离出的mRNA的卵母细胞表达了新的钾电流,这些电流在注射后的第二天出现,并表现出时间和电压依赖性激活,随后是缓慢失活。它们在-90 mV开始激活,半激活电压(V1/2)为-30 mV;其激活时间常数(在+60 mV时)为0.11秒,而失活时间常数为1.92秒,V1/2 = -44.6 mV。这种钾电流被钾通道阻滞剂四乙铵(TEA)、4-氨基吡啶(4AP)以及钡离子(Ba(2+))、奎宁、奎尼丁、蝎毒素、树眼镜蛇毒素-1、辣椒素、玛格毒素和地尔硫卓有效阻断。这些结果表明,这种电流是蓝氏贾第鞭毛虫电压门控钾通道在非洲爪蟾卵母细胞中异源表达的结果。