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Spatial expression pattern of serine proteases in the blood fluke Schistosoma mansoni determined by fluorescence RNA in situ hybridization
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SYSNO ASEP 0543094 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Spatial expression pattern of serine proteases in the blood fluke Schistosoma mansoni determined by fluorescence RNA in situ hybridization Author(s) Ulrychová, Lenka (UOCHB-X) ORCID, RID
Ostašov, P. (CZ)
Chanová, M. (CZ)
Mareš, Michael (UOCHB-X) RID, ORCID
Horn, Martin (UOCHB-X) RID, ORCID
Dvořák, J. (CZ)Article number 274 Source Title Parasites & Vectors. - : BioMed Central - ISSN 1756-3305
Roč. 14, č. 1 (2021)Number of pages 14 s. Language eng - English Country GB - United Kingdom Keywords blood fluke ; fluorescence RNA in situ hybridization ; mRNA detection ; platyhelminthes ; Schistosoma mansoni ; serine proteases ; transcript OECD category Biochemistry and molecular biology R&D Projects NV18-05-00345 GA MZd - Ministry of Health (MZ) GA19-17269S GA ČR - Czech Science Foundation (CSF) 8J19AT036 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 000657776800003 EID SCOPUS 85106735639 DOI 10.1186/s13071-021-04773-8 Annotation Background: The blood flukes of genus Schistosoma are the causative agent of schistosomiasis, a parasitic disease that infects more than 200 million people worldwide. Proteases of schistosomes are involved in critical steps of host–parasite interactions and are promising therapeutic targets. We recently identified and characterized a group of S1 family Schistosoma mansoni serine proteases, including SmSP1 to SmSP5. Expression levels of some SmSPs in S. mansoni are low, and by standard genome sequencing technologies they are marginally detectable at the method threshold levels. Here, we report their spatial gene expression patterns in adult S. mansoni by the high-sensitivity localization assay. Methodology: Highly sensitive fluorescence in situ RNA hybridization (FISH) was modified and used for the localization of mRNAs encoding individual SmSP proteases (including low-expressed SmSPs) in tissues of adult worms. High sensitivity was obtained due to specifically prepared tissue and probes in combination with the employment of a signal amplification approach. The assay method was validated by detecting the expression patterns of a set of relevant reference genes including SmCB1, SmPOP, SmTSP-2, and Sm29 with localization formerly determined by other techniques. Results: FISH analysis revealed interesting expression patterns of SmSPs distributed in multiple tissues of S. mansoni adults. The expression patterns of individual SmSPs were distinct but in part overlapping and were consistent with existing transcriptome sequencing data. The exception were genes with significantly low expression, which were also localized in tissues where they had not previously been detected by RNA sequencing methods. In general, SmSPs were found in various tissues including reproductive organs, parenchymal cells, esophagus, and the tegumental surface. Conclusions: The FISH-based assay provided spatial information about the expression of five SmSPs in adult S. mansoni females and males. This highly sensitive method allowed visualization of low-abundantly expressed genes that are below the detection limits of standard in situ hybridization or by RNA sequencing. Thus, this technical approach turned out to be suitable for sensitive localization studies and may also be applicable for other trematodes. The results suggest that SmSPs may play roles in diverse processes of the parasite. Certain SmSPs expressed at the surface may be involved in host–parasite interactions. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Viktorie Chládková, Tel.: 232 002 434 Year of Publishing 2022 Electronic address https://doi.org/10.1186/s13071-021-04773-8
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