Abstract
BackgroundAquaporins (AQPs) are a family of integral membrane channel proteins that facilitate the transport of water and other small solutes across cell membranes. AQPs appear to play crucial roles in parasite survival and represent possible drug targets for novel intervention strategy. In this work, we investigated the tissue distribution and biological roles of an aquaporin TcAQP1 in the neglected parasitic nematode Toxocara canis.MethodsRecombinant C-terminal hydrophilic domain of AQP1 of T. canis (rTcAQP1c) and polyclonal antibody against rTcAQP1c were produced to analyse the tissue expression of native TcAQP1 in adult (female and male) worms using an immunohistochemical approach. RNA interference (RNAi), quantitative real-time PCR (qRT-PCR) and nematocidal assays were performed to investigate the functional roles of TcAQP1 in the adult stage of T. canis.ResultsImmunofluorescence analysis showed that TcAQP1 was localised predominantly in the epithelial linings of the reproductive tract and basolateral membrane of the intestine in the adult stage (female and male) of T. canis, indicating important roles in reproduction, nutrient absorption and/or osmoregulation. Treatment with silencing RNA for 24 h resulted in a significant reduction of Tc-aqp-1 mRNA level in adult T. canis, though no phenotypical change was observed. The efficient gene knockdown compromised the nematocidal activity of albendazole in vitro, suggesting the role of TcAQP1 in drug uptake.ConclusionsThe findings of this study provide important information about tissue expression and functional roles of TcAQP1 protein in adult T. canis. Understanding the biological functions of this protein in other developmental stages of T. canis and related parasitic nematodes would contribute to the discovery of novel diagnostic or anthelmintic targets.
Highlights
Aquaporins (AQPs) are a family of integral membrane channel proteins that facilitate the transport of water and other small solutes across cell membranes
Specific polyclonal antibody against rTcAQP1c peptide The C-terminal hydrophilic domain of aquaporin 1 protein of Toxocara canis (TcAQP1) was expressed in BL21 (DE3) cells as a 6× His-GST fusion protein (Fig. 1a)
Quantitative real‐time polymerase chain reaction (PCR) After soaking for 24 h, the efficacy of gene knockdown was determined by comparing the relative mRNA levels of Tc-aqp-1 between the worms treated with silencing and non-silencing RNAs. quantitative real-time PCR (qRT-PCR) was performed as described previously [30] to confirm the efficiency of gene knockdown
Summary
Aquaporins (AQPs) are a family of integral membrane channel proteins that facilitate the transport of water and other small solutes across cell membranes. AQPs appear to play crucial roles in parasite survival and represent possible drug targets for novel intervention strategy. Aquaporin 1, an integral membrane channel protein facilitating the transport of water, was first identified in human erythrocytes [1, 2]. Schistosoma mansoni [20,21,22,23] These findings have been raising interest in the proposition that AQPs might represent novel therapeutic targets for parasitic diseases [24,25,26,27]. A better understanding of this area in parasitic nematodes would contribute to the discovery of novel anthelmintic targets
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