Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites

0519756 - BC 2020 RIV GB eng J - Journal Article
Kosakyan, Anush - Alama-Bermejo, Gema - Bartošová-Sojková, Pavla - Born-Torrijos, Ana - Šíma, Radek - Nenarokova, Anna - Eszterbauer, E. - Bartholomew, J. - Holzer, Astrid S.
Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites.
Scientific Reports. Roč. 9, OCT 21 2019 (2019), č. článku 15073. ISSN 2045-2322. E-ISSN 2045-2322
R&D Projects: GA ČR(CZ) GBP505/12/G112; GA ČR(CZ) GP14-28784P; GA MŠMT(CZ) LTAUSA17201
EU Projects: European Commission(XE) 634429 - ParaFishControl
Institutional support: RVO:60077344
Keywords : real-time pcr * sphaerospora-molnari myxozoa * proliferative kidney-disease * ceratomyxa-shasta myxozoa * fresh-water polychaete * rainbow-trout * myxobolus-cerebralis * housekeeping genes * rt-pcr * differential expression
OECD category: Veterinary science
Impact factor: 3.998, year: 2019
Method of publishing: Open access
https://www.nature.com/articles/s41598-019-51479-0.pdf

Myxozoans (Cnidaria: Myxozoa) are an extremely diversified group of endoparasites some of which are causative agents of serious diseases in fish. New methods involving gene expression studies have emerged over the last years to better understand and control myxozoan diseases. Quantitative RTPCR is the most extensively used approach for gene expression studies. However, the accuracy of the results depends on the normalization of the data to reference genes. We studied the expression of eight commonly used reference genes, adenosylhomocysteinase (AHC1), beta actin (ACTB), eukaryotic translation elongation factor 2 (EF2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase 1 (HPRT1), DNA-directed RNA polymerase II (RPB2), 18S ribosomal RNA (18S), 28S ribosomal RNA (28S) across different developmental stages of three myxozoan species, Sphaerospora molnari, Myxobolus cerebralis and Ceratonova shasta, representing the three major myxozoan linages from the largest class Myxosporea. The stable reference genes were identified using four algorithms: geNorm, NormFinder, Bestkeeper and Delta Cq method. Additionally, we analyzed transcriptomic data from S. molnari proliferative and spore-forming stages to compare the relative amount of expressed transcripts with the most stable reference genes suggested by RT-qPCR. Our results revealed that GAPDH and EF2 are the most uniformly expressed genes across the different developmental stages of the studied myxozoan species.
Permanent Link: http://hdl.handle.net/11104/0304748