Ponce Arturo, Ogazon Del Toro Alejandro, Jimenez Lidia, Eligio-Garcia Leticia, Jimenez-Cardoso Enedina
Department of Physiology, Biophysics and Neurosciences. Center for Research and Advanced Studies, Mexico City, Mexico.
Parasitology Research Laboratory, Children Hospital of México "Federico Gomez", Mexico City, México.
Physiol Rep. 2019 Jun;7(11):e14029. doi: 10.14814/phy2.14029.
Giardia lamblia is one of the most important worldwide causes of intestinal infections, yet little is known about its cellular physiology, especially the diversity of ionic channels that this parasite expresses. In this work, we show that injection of mRNA isolated from trophozoites of Giardia, into Xenopus laevis oocytes, induces expression of three types of chloride currents (here referred to as ICl-G1, ICl-G2, and ICl-G3), which have different biophysical and pharmacological properties. ICl-G1 currents show inward rectification and voltage dependence are enhanced by hypotonicity, show a selectivity sequence of (I > Br > Cl > F), and are inhibited by NPPB, DIDS, SITS, 9AC, DPC, and Zinc. These findings suggest that ICl-G1 is the result of expression of chloride channels related to ClC2. ICl-G2 currents show outward rectification and are dependent of intracellular calcium, its selectivity sequence is (Cl > Br > I > F) and are inhibited by NPPB, DIDS, SITS, 9AC, DPC, niflumic acid, tannic acid, and benzbromarone. These findings suggest that they are produced by calcium dependent chloride channels (CaCC). The third type of currents (ICl-G3) appears only after a hypoosmotic challenge, and has similar properties to those described for ICl-swell, such as outward rectification, instant activation, and slow inactivation at large depolarizing voltages. They were blocked by NPPB, DIDS, 9AC, NIf, DCPIB, and tamoxifen. Our results indicate that Giardia intestinalis has at least three types of anion conductances.
蓝氏贾第鞭毛虫是全球肠道感染的最重要病因之一,但对其细胞生理学,尤其是该寄生虫所表达的离子通道多样性却知之甚少。在本研究中,我们发现,将从蓝氏贾第鞭毛虫滋养体中分离的mRNA注射到非洲爪蟾卵母细胞中,可诱导三种类型氯离子电流(以下称为ICl-G1、ICl-G2和ICl-G3)的表达,这些电流具有不同的生物物理和药理学特性。ICl-G1电流表现为内向整流,低渗可增强其电压依赖性,其选择性顺序为(I>Br>Cl>F),并被NPPB、DIDS、SITS、9AC、DPC和锌抑制。这些发现表明,ICl-G1是与ClC2相关的氯离子通道表达的结果。ICl-G2电流表现为外向整流,依赖细胞内钙,其选择性顺序为(Cl>Br>I>F),并被NPPB、DIDS、SITS、9AC、DPC、氟尼酸、鞣酸和苯溴马隆抑制。这些发现表明,它们是由钙依赖性氯离子通道(CaCC)产生的。第三种类型的电流(ICl-G3)仅在低渗刺激后出现,其特性与ICl-swell相似,如外向整流、瞬间激活以及在大去极化电压下缓慢失活。它们被NPPB、DIDS、9AC、NIf、DCPIB和他莫昔芬阻断。我们的结果表明,肠道贾第虫至少有三种类型的阴离子电导。