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Open AccessArticle

Comparative Proteomics Analysis of Anisakis simplex s.s.—Evaluation of the Response of Invasive Larvae to Ivermectin

1
Department of Biochemistry, Faculty of Biology and Biotechnology, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, Poland
2
Department of Food Technology, Marine Research Institute (IIM), Spanish National Research Council (CSIC), 36-208 Vigo, Spain
*
Authors to whom correspondence should be addressed.
Genes 2020, 11(6), 710; https://doi.org/10.3390/genes11060710
Received: 28 May 2020 / Revised: 19 June 2020 / Accepted: 25 June 2020 / Published: 26 June 2020
(This article belongs to the Special Issue Novel Omics Studies on Anisakid Nematodes)
Ivermectin (IVM), an antiparasitic drug, has a positive effect against Anisakis simplex s.s. infection and has been used for the treatment and prevention of anisakiasis in humans. However, the molecular mechanism of action of IVM on A. simplex s.s. remains unknown. Herein, tandem mass tag (TMT) labeling and extensive liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis were used to identify the effect of IVM on the proteome of A. simplex s.s. in vitro. During the study, 3433 proteins, of which 1247 had at least two protein unique peptides, were identified. Comparative proteomics analysis revealed that 59 proteins were differentially regulated (DRPs) in IVM-treated larvae, of which 14 proteins were upregulated and 38 were downregulated after 12 h of culture, but after 24 h, 12 proteins were upregulated and 22 were downregulated. The transcription level of five randomly selected DRPs was determined by real-time PCR as a supplement to the proteomic data. The functional enrichment analysis showed that most of the DRPs were involved in oxidoreductase activity, immunogenicity, protein degradation, and other biological processes. This study has, for the first time, provided comprehensive proteomics data on A. simplex s.s. response to IVM and might deliver new insight into the molecular mechanism by which IVM acts on invasive larvae of A. simplex s.s. View Full-Text
Keywords: parasitic nematode; anisakiasis; Anisakis simplex; proteome; mass spectrometry; real-time PCR; antiparasitic drugs parasitic nematode; anisakiasis; Anisakis simplex; proteome; mass spectrometry; real-time PCR; antiparasitic drugs
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    Doi: 10,5281/zenodo.3862579
    Description: Figure S1: The visualization of SDS-polyacrylamide electrophoresis. Protein extracts were separated and evaluated on 12% (v/v) polyacrylamide gels (20:1) with a stacking gel of 4% polyacrylamide. Gel was silver stained. Lane 1—molecular weight marker; lanes 2‒4—control samples; lanes 5‒7—IVM-treated samples for 12 h; lanes 8‒10—IVM-treated samples for 24 h; File S1: Data repository with all identified proteins for Anisakis simplex s.s. proteins cultured with and without ivermectin for 12 h and 24 h; File S2: Data repository with all differentially regulated proteins (DRPs) in Anisakis simplex s.s., cultured with and without ivermectin for 12 h and 24 h. Additionally, all proteins identified with at least two protein-unique peptides are presented; File S3: Data repository with detailed functional enrichment analysis., Table S1: The primers of ivermectin-induced DRPs used for real-time PCR assay for verification of mRNA levels in Anisakis simplex s.s. invasive larvae.
MDPI and ACS Style

Polak, I.; Łopieńska-Biernat, E.; Stryiński, R.; Mateos, J.; Carrera, M. Comparative Proteomics Analysis of Anisakis simplex s.s.—Evaluation of the Response of Invasive Larvae to Ivermectin. Genes 2020, 11, 710.

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