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Effect of Ichthyophthirius multifiliis on gene expression after treatment with magnolol in vivo

ABSTRACT: To investigate the antiparasitic mechanism of magnolol, we performed RNA-seq analysis to probe the significantly regulated genes after treatemnt by magnolol. We then performed gene expression profiling analysis using data obtained from RNA-seq of 13 independent parasite samples collected form various treatment goups.

ORGANISM(S): Ichthyophthirius multifiliis

PROVIDER: GSE214083 | GEO | 2023/09/26

REPOSITORIES: GEO

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Project description:Schistosoma mansoni is the major causative agent of schistosomiasis in the Americas. This parasite takes advantage from host signaling molecules such as cytokines and hormones to complete its development inside the host. TNF-Î± is the most important cytokine involved in the inflammatory response when cercaria, the infective stage, penetrates the human skin and a severe inflammatory response is started. In this work the authors describe the complete sequence of a possible TNF-Î± receptor in S. mansoni and detect that the receptor is most highly expressed in cercaria among all life cycle stages. In an attempt to mimic the situation at the site of skin penetration cercariae have been mechanically transformed in vitro into schistosomula and immediately exposed to human TNF-Î± . Exposure of these early schistosomula to the human hormone caused a large-scale change in the expression of parasite genes. Exposure of adult worms to human TNF-Î± caused gene expression changes as well, although the set of parasite altered genes was entirely different from that of schistosomula. This work increases the number of known signaling pathways of the parasite, and opens new perspectives into understanding the molecular components of TNF-Î± response as well as possibly interfering with parasite-host interaction. A 4 x 44k microarray platform was designed with the same probe set described by our group in a previous publication (PMID: 17517391). 230 ng of RNA from schistosomula treated with TNF and its control were used for the hybridization. RNA amplification and labeling kit (Agilent Technologies) were used according to manufacturerÂ´s instructions. Each sample was separately labeled with either Cy3 or Cy5 and 825 ng cRNA from each amplification were used for hybridization a control sample was combined with a treated sample and hybridized. Washing and scanning procedures were according to the manufacturerÂ´s instructions using GenePix 4000B scanner (Molecular Devices, Sunnyvale, CA, USA). Data was extracted using Feature Extraction software (Agilent Technologies). Low intensity data points were filtered out according to Feature Extraction software criteria, which essentially determine those points that are significantly below the average background signal of the array. Total intensity data from each experiment were normalized by Quantiles Normalization Method, excluding positive and negative external controls. Significance Analysis of Microarray (SAM) was used as to find differentially expressed genes. z-score transformation of normalized data was performed and SAM two class tests were applied; genes were considered as significantly differentially expressed at q-value â¤ 0.05. Hierarchical clustering of selected genes was generated using Spotfire Decision Site software (TIBCO Software Inc., Palo Alto, CA, USA). For a gene that was represented in the array by multiple probes, we picked a single representative probe by selecting the probe with the highest absolute value of the Log2 ratio between intensities of TNF-Î± Treated/Control.

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Effect of Ichthyophthirius multifiliis on gene expression after treatment with magnolol in vivo

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