Downregulated RPS-30 in Angiostrongylus cantonensis L5 plays a defensive role against damage due to oxidative stress
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Parasites & Vectors Open Access
RESEARCH
Downregulated RPS‑30 in Angiostrongylus cantonensis L5 plays a defensive role against damage due to oxidative stress Wei‑Wei Sun2, Xiu‑Mei Yan3, Qing Shi1, Yuan‑Jiao Zhang1, Jun‑Ting Huang4, Hui‑Cong Huang1*, Hong‑Fei Shi5* and Bao‑Long Yan1*
Abstract Background: Eosinophilic meningitis, caused by fifth-stage larvae of the nematode (roundworm) Angiostrongylus cantonensis, is mainly attributed to the contribution of eosinophils to tissue inflammatory responses in helminthic infections. Eosinophils are associated with the killing of helminths via peroxidative oxidation and hydrogen peroxide generated by the dismutation of superoxide produced during respiratory bursts. In contrast, when residing in the host with high level of eosinophils, helminthic worms have evolved to attenuate eosinophil-mediated tissue inflammatory responses for their survival in the hosts. In a previous study we demonstrated that the expression of the A. cantonensis RPS 30 gene (Acan-rps-30) was significantly downregulated in A. cantonensis L5 roundworms residing in cerebrospinal fluid with a high level of eosinophils. Acan-RPS-30 is a protein homologous to the human Fau protein that plays a proapoptotic regulatory role and may function in protecting worms from oxidative stress. Methods: The isolation and structural characterization of Acan-RPS-30 were performed using rapid amplification of cDNA ends (RACE), genome walking and bioinformatics. Quantitative real-time-PCR and microinjection were used to detect the expression patterns of Acan-rps-30. Feeding RNA interference (RNAi) was used to knockdown the apop‑ tosis gene ced-3. Microinjection was performed to construct transgenic worms. An oxidative stress assay was used to determine the functions of Acan-RPS-30. Results: Our results showed that Acan-RPS-30 consisted of 130 amino acids. It was grouped into clade V with C. elegans in the phylogenetic analysis. It was expressed ubiquitously in worms and was downregulated in both L5 larvae and adult A. cantonensis. Worms expressing pCe-rps30::Acan-rps-30::rfp, with the refractile “button-like” apop‑ totic corpses, were susceptible to oxidative stress. Apoptosis genes ced-3 and ced-4 were both upregulated in the transgenic worms. The phenotype susceptible to oxidative stress could be converted with a ced-3 defective mutation and RNAi. rps-30−/− mutant worms were resistant to oxidative stress, with ced-3 and ced-4 both downregulated. The oxidative stress-resistant phenotype could be rescued and inhibited by through the expression of pCe-rps30::Acan-rps30::rfp in rps-3−/− mutant worms.
*Correspondence: [email protected]; [email protected]; 1982ybllog@163. com † Wei-Wei Sun and Xiu-Mei Yan contribute equally to this work 1 Department of Parasitology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, Zhejiang, PR China 5 Henan Provincial Engineering Laboratory of Insects Bio‑reactor, China‑UK‑NYNU‑RRes Joint Laboratory of Insect Biology, Nanyang Normal University, Nanyang 473061, PR Ch
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