Abstract：We explored the roles of aquaporins (AQPs) of Echinococcus granulosus and E. multilocularis in the formation of hydatid fluid, which may provide a foundation for better understanding the mechanism of formation of hydatid fluid and selecting new targets of chemotherapy for echinococcosis. AQPs genes of E. granulosus (EgAQPs) and E. multilocularis (EmAQPs) were cloned by the method of RT-PCR. Heterologous expression vectors of AQPs were constructed and heterogeneously expressed in Xenopus laevis oocytes in order to identify the water channel activities of these AQPs. The characteristics and differences of transmembrane regions between EgAQPs and EmAQPs were analyzed by bioinformatics method. In addition, multiple sequence alignment was carried out to analyze the structure difference between human AQPs and AQPs of Echinococcus spp. Results showed that two EgAQPs genes (EgAQP4 and EgAQP9) and two EmAQPs genes (EmAQP4 and EmAQP9) were isolated successfully by the method of RT-PCR. The swelling rate (V/V0) of AQPs cRNA-injected oocytes was not significantly different from DEPC water-injected oocytes in control (F=1.143, P＞0.05), as well as osmotic water permeability coefficient (F=1.416, P>0.05). These four AQPs genes could not perform their water channel activities in X. laevis oocyte. Results of transmembrane regions prediction and multiple sequence alignment showed that compared with the conventional AQPs, although N and C termins of EgAQP4 and EmAQP4 were located in cytoplasm, the number of transmembrane regions was four, and NPA motifs were replaced with NPM and NPT; although NPA motifs were conserved in EgAQP9 and EmAQP9, the number of transmembrane regions was five, and N termin was extracellular, but C termin was cytoplasmic. AQPs of Echinococcus could not perform their water channel activities in X. laevis oocyte may be caused by these structure differences between AQPs of Echinococcus and human AQPs. Although AQP encoding genes were identified in the genome of E. granulosus and E. multilocularis, the water channel activity could not be found in the functional verification test of these AQPs genes. All these results may suggest that neither water can enter the germinal layer to cause vesicle lysis nor can water be transported through these AQPs so that the accumulation of hydatid fluid can increase the volume of hydatid cyst or vesicles.
刘许诺,王芬,吴宏烨,李锴,范俊杰,叶彬. 两种棘球绦虫的水通道蛋白基因克隆及功能特性的比较[J]. 中国人兽共患病学报, 2018, 34(6): 501-508.
LIU Xu-nuo, WANG Fen, WU Hong-ye, LI Kai, FAN Jun-jie, YE Bin. Comparison of gene clone and functional characteristics of aquaporins of Echinococcus granulosus and Echinococcus multilocularis. Chinese Journal of Zoonoses, 2018, 34(6): 501-508.
[1] Wang Q, Huang Y, Huang L, et al.Review of risk factors for human echinococcosis prevalence on the Qinghai-Tibet Plateau, China: a prospective for control options[J].Infect Dis Poverty, 2014, 3(1): 3.DOI: 10.1186/2049-9957-3-3 [2] Sezgin O, Altintas E, Saritas U, et al.Hepatic alveolar echinococcosis: clinical and radiologic features and endoscopic management[J].J Clin Gastroenterol, 2005, 39(2): 160-167.DOI:10.1097/01.mcg.0000150242.72499.22 [3] Hemphill A, Mueller J.Alveolar and cystic echinococcosis: towards novel chemotherapeutical treatment options[J].J Helminthol, 2009, 83(2): 99-111.DOI: 10.1017/S0022149-X0928936X [4] Kern P.Clinical features and treatment of alveolar echinococcosis[J].Curr Opin Infect Dis, 2010, 23(5): 505-512.DOI: 10.1097/QCO.0b013e32833d7516 [5] 张宏伟, 彭心宇.中西药联合阿苯达唑治疗棘球蚴病研究进展[J].中国病原生物学杂志, 2009, 4(9): 705-708. [6] Reuter S, Jensen B, Buttenschoen K, et al.Benzimidazoles in the treatment of alveolar echinococcosis: a comparative study and review of the literature[J].J Antimicrob Chemother, 2000, 46(3): 451-456.DOI: 10.1093/jac/46.3.451 [7] 余森海.棘球蚴病防治研究的国际现状和对我们的启示[J].中国寄生虫学与寄生虫病杂志, 2008, 26(4): 241-244. [8] 隋海心, 任罡.水分子通道蛋白的结构与功能[J].化学进展, 2004, 16(2): 145-152. [9] Abascal F, Irisarri L, Zardoya R.Diversity and evolution of membrane intrinsic proteins[J].Biochim Biophys Acta, 2014, 1840(5): 1468-1481.DOI: 10.1016/j.bbagen.2013.12.001 [10] Wang Y, Tajkhorshid E.Molecular mechanisms of conduction and selectivity in aquaporin water channels[J].J Nutr, 2007, 137(6 suppl 1): 1509-1515 [11] Ishibashi K.Aquaporin subfamily with unusual NPA boxes[J].Biochim Biophys Acta, 2006, 1758(8): 989-993.DOI: 10.1016/j.bbamem.2006.02.024 [12] Gomes D, Agasse A, Thiebaud P, et al.Aquaporins are multifunctional water and solute transporters highly divergent in living organisms[J].Biochim Biophys Acta, 2009, 1788(6): 1213-1228.DOI: 10.1016/j.bbamem.2009.03.009 [13] Beitz E.Aquaporin water and solute channels from malaria parasites and other pathogenic protozoa[J].Chem Med Chem, 2006, 1(6): 587-592.DOI: 10.1002.cmdc.200500105 [14] Song J, Mak E, Wu BH, et al.Parasite aquaporins: Current developments in drug facilitation and resistance[J].Biochim Biophysica Acta, 2014, 1840(5): 1566-1573.DOI: 10.1016/j.bbagen.2013.10.014 [15] Reisin IL, Pavisicdefal C.Membrane permeability of secondary hydatid cysts of Echinococcus granulosus.Determination of the water diffusional and osmotic permeability cofficients through a syncytial membrane[J].Mol Biochem Parasitol, 1984, 12(1):101-116.DOI: 10.1016/0166-6851(84)90048-3 [16] Zheng HJ, Zhang WB, Zhang L, et al.The genome of the hydatid tapeworm Echinococcus granulosus[J].Nat Genet, 2013, 45(10): 1168-1175.DOI: 10.1038/ng.2757 [17] Zhang RB, Verkman AS.Water and urea permeability properties of Xenopus oocytes: expression of mRNA from toad urinary bladder[J].Am J Physiol, 1991, 260 (1): 26-34.DOI: 10.1152/ajpcell.1991.260.1.C26 [18] Torgerson PR, Keller K, Magnotta M, et al.The global burden of alveolar echinococcosis[J].PLoS Negl Trop Dis, 2010, 4(6): e722-e722.DOI: 10.1371/journal.pntd.0000722 [19] 叶尔江·苏里唐, 江莉, 柴君杰.我国棘球蚴病防治研究进展[J].中国寄生虫学与寄生虫病学杂志, 2000, 18 (13): 179-181. [20] 温浩, 吐尔干艾力·阿吉, 邵英梅, 等.棘球蚴病防治成就及面临的挑战[J].中国寄生虫学与寄生虫病杂志, 2015, 33 (6): 466-471. [21] 李红卫, 袁芳, 李燕兵, 等.昆明种小鼠棘球蚴病感染动物模型的建立[J].宁夏医科大学学报, 2011, 33(5): 411-413. [22] Geadkaew A, Bulow JV, Beitz E, et al.Bi-functionality of Opisthorchis viverrini aquaporins[J].Biochimie, 2015, 108: 149-159.DOI: 10.1016/j.biochi.2014.11.013 [23] Huang CG, Lamitina T, Agre P, et al.Functional analysis of the aquaporin gene family in Caenorhabditis elegans[J].Am J Physiol Cell Physiol, 2007, 292 (5): C1867-C1873.DOI: 10.1152/ajpcell.00514.2006 [24] Geadkaew A, Von BJ, Beitz E, et al.Functional analysis of novel aquaporins from Fasciola gigantica[J].Mol Biochem Parasitol, 2011, 175(2): 144-153.DOI: 10.1016/j.molbiopara.2010.10.010 [25] Figarella K, Uzcategui NL, Zhou Y, et al.Biochemical characterization of Leishmania major aquaglyceroporin LmAQP1: possible role in volume regulation and osmotaxis[J].Mol Microbiol, 2007, 65(4): 1006-1017.DOI: 10.1111/j.1365-2958.2007.05845x [26] Hedfalk K, Pettersson N, Oberg f, et al.Production, characterization and crystallization of the Plasmodium falciparum aquaporin[J].Protein Expr Purif, 2008, 59(1): 69-78.DOI: 10.1016/j.pep.2008.01.004
2018, Vol. 34 
