Department of Veterinary and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg C, Denmark.
INRAE, Université de Tours, ISP, Nouzilly, France.
PLoS Pathog. 2021 Feb 5;17(2):e1008982. doi: 10.1371/journal.ppat.1008982. eCollection 2021 Feb.
In the absence of efficient alternative strategies, the control of parasitic nematodes, impacting human and animal health, mainly relies on the use of broad-spectrum anthelmintic compounds. Unfortunately, most of these drugs have a limited single-dose efficacy against infections caused by the whipworm, Trichuris. These infections are of both human and veterinary importance. However, in contrast to a wide range of parasitic nematode species, the narrow-spectrum anthelmintic oxantel has a high efficacy on Trichuris spp. Despite this knowledge, the molecular target(s) of oxantel within Trichuris is still unknown. In the distantly related pig roundworm, Ascaris suum, oxantel has a small, but significant effect on the recombinant homomeric Nicotine-sensitive ionotropic acetylcholine receptor (N-AChR) made up of five ACR-16 subunits. Therefore, we hypothesized that in whipworms, a putative homolog of an ACR-16 subunit, can form a functional oxantel-sensitive receptor. Using the pig whipworm T. suis as a model, we identified and cloned a novel ACR-16-like subunit and successfully expressed the corresponding homomeric channel in Xenopus laevis oocytes. Electrophysiological experiments revealed this receptor to have distinctive pharmacological properties with oxantel acting as a full agonist, hence we refer to the receptor as an O-AChR subtype. Pyrantel activated this novel O-AChR subtype moderately, whereas classic nicotinic agonists surprisingly resulted in only minor responses. We observed that the expression of the ACR-16-like subunit in the free-living nematode Caenorhabditis elegans conferred an increased sensitivity to oxantel of recombinant worms. We demonstrated that the novel Tsu-ACR-16-like receptor is indeed a target for oxantel, although other receptors may be involved. These finding brings new insight into the understanding of the high sensitivity of whipworms to oxantel, and highlights the importance of the discovery of additional distinct receptor subunit types within Trichuris that can be used as screening tools to evaluate the effect of new synthetic or natural anthelmintic compounds.
在缺乏有效替代策略的情况下,控制影响人类和动物健康的寄生线虫主要依赖于广谱驱虫化合物的使用。不幸的是,这些药物中的大多数对鞭虫引起的感染的单剂量疗效有限。这些感染对人类和兽医都很重要。然而,与广泛的寄生线虫种类不同,窄谱驱虫药奥沙苯达唑对线虫属具有很高的疗效。尽管有这些知识,但奥沙苯达唑在鞭虫中的分子靶标仍然未知。在远缘相关的猪蛔虫中,奥沙苯达唑对由五个 ACR-16 亚基组成的重组同源 Nicotine-sensitive ionotropic acetylcholine receptor (N-AChR) 有很小但显著的影响。因此,我们假设在鞭虫中,假定的 ACR-16 亚基同源物可以形成功能性的奥沙苯达唑敏感受体。我们使用猪鞭虫 T. suis 作为模型,鉴定并克隆了一种新型 ACR-16 样亚基,并成功地在非洲爪蟾卵母细胞中表达了相应的同源通道。电生理学实验表明,该受体具有独特的药理学特性,奥沙苯达唑作为完全激动剂,因此我们将该受体称为 O-AChR 亚型。噻嘧啶适度激活这种新型 O-AChR 亚型,而经典的烟碱激动剂出人意料地仅产生较小的反应。我们观察到,在自由生活的线虫秀丽隐杆线虫中表达 ACR-16 样亚基赋予重组蠕虫对奥沙苯达唑的敏感性增加。我们证明了新型 Tsu-ACR-16 样受体确实是奥沙苯达唑的靶标,尽管可能涉及其他受体。这些发现为理解鞭虫对线虫属的高度敏感性提供了新的见解,并强调了在 Trichuris 中发现额外独特的受体亚基类型的重要性,这些类型可以用作筛选工具来评估新的合成或天然驱虫化合物的效果。