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

Autores
Kosakyan, Anush; Alama Bermejo, Gema; Bartošová-Sojková, Pavla; Born-Torrijos, Ana; Šíma, Radek; Nenarokova, Anna; Eszterbauer, Edit; Bartholomew, Jerri; Holzer, Astrid Sybylle
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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 RT-PCR 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 ΔCq 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.
Fil: Kosakyan, Anush. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Alama Bermejo, Gema. Universidad Nacional del Comahue. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". - Provincia de Río Negro. Ministerio de Agricultura, Ganadería y Pesca. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Centro Nacional Patagónico. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni"; Argentina. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. State University of Oregon; Estados Unidos
Fil: Bartošová-Sojková, Pavla. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Born-Torrijos, Ana. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Šíma, Radek. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Nenarokova, Anna. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. University of South Bohemia; República Checa
Fil: Eszterbauer, Edit. Hungarian Academy Of Sciences; Hungría
Fil: Bartholomew, Jerri. State University of Oregon; Estados Unidos
Fil: Holzer, Astrid Sybylle. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Materia
MYXOZOA
REFERENCE GENES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/117400
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasitesKosakyan, AnushAlama Bermejo, GemaBartošová-Sojková, PavlaBorn-Torrijos, AnaŠíma, RadekNenarokova, AnnaEszterbauer, EditBartholomew, JerriHolzer, Astrid SybylleMYXOZOAREFERENCE GENEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Myxozoans (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 RT-PCR 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 ΔCq 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.Fil: Kosakyan, Anush. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Alama Bermejo, Gema. Universidad Nacional del Comahue. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". - Provincia de Río Negro. Ministerio de Agricultura, Ganadería y Pesca. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Centro Nacional Patagónico. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni"; Argentina. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. State University of Oregon; Estados UnidosFil: Bartošová-Sojková, Pavla. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Born-Torrijos, Ana. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Šíma, Radek. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaFil: Nenarokova, Anna. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. University of South Bohemia; República ChecaFil: Eszterbauer, Edit. Hungarian Academy Of Sciences; HungríaFil: Bartholomew, Jerri. State University of Oregon; Estados UnidosFil: Holzer, Astrid Sybylle. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República ChecaNature Publishing Group2019-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/117400Kosakyan, Anush; Alama Bermejo, Gema; Bartošová-Sojková, Pavla; Born-Torrijos, Ana; Šíma, Radek; et al.; Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites; Nature Publishing Group; Scientific Reports; 9; 1; 12-2019; 1-142045-2322CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-019-51479-0info:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/s41598-019-51479-0info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:42:12Zoai:ri.conicet.gov.ar:11336/117400instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:42:12.697CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
title Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
spellingShingle Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
Kosakyan, Anush
MYXOZOA
REFERENCE GENES
title_short Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
title_full Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
title_fullStr Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
title_full_unstemmed Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
title_sort Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites
dc.creator.none.fl_str_mv Kosakyan, Anush
Alama Bermejo, Gema
Bartošová-Sojková, Pavla
Born-Torrijos, Ana
Šíma, Radek
Nenarokova, Anna
Eszterbauer, Edit
Bartholomew, Jerri
Holzer, Astrid Sybylle
author Kosakyan, Anush
author_facet Kosakyan, Anush
Alama Bermejo, Gema
Bartošová-Sojková, Pavla
Born-Torrijos, Ana
Šíma, Radek
Nenarokova, Anna
Eszterbauer, Edit
Bartholomew, Jerri
Holzer, Astrid Sybylle
author_role author
author2 Alama Bermejo, Gema
Bartošová-Sojková, Pavla
Born-Torrijos, Ana
Šíma, Radek
Nenarokova, Anna
Eszterbauer, Edit
Bartholomew, Jerri
Holzer, Astrid Sybylle
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv MYXOZOA
REFERENCE GENES
topic MYXOZOA
REFERENCE GENES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv 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 RT-PCR 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 ΔCq 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.
Fil: Kosakyan, Anush. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Alama Bermejo, Gema. Universidad Nacional del Comahue. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". - Provincia de Río Negro. Ministerio de Agricultura, Ganadería y Pesca. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni". Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Centro Nacional Patagónico. Centro de Investigación Aplicada y Transferencia Tecnológica en Recursos Marinos "Almirante Storni"; Argentina. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. State University of Oregon; Estados Unidos
Fil: Bartošová-Sojková, Pavla. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Born-Torrijos, Ana. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Šíma, Radek. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
Fil: Nenarokova, Anna. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa. University of South Bohemia; República Checa
Fil: Eszterbauer, Edit. Hungarian Academy Of Sciences; Hungría
Fil: Bartholomew, Jerri. State University of Oregon; Estados Unidos
Fil: Holzer, Astrid Sybylle. Academy of Sciences of the Czech Republic. Biology Centre. Institute of Parasitology; República Checa
description 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 RT-PCR 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 ΔCq 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.
publishDate 2019
dc.date.none.fl_str_mv 2019-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/117400
Kosakyan, Anush; Alama Bermejo, Gema; Bartošová-Sojková, Pavla; Born-Torrijos, Ana; Šíma, Radek; et al.; Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites; Nature Publishing Group; Scientific Reports; 9; 1; 12-2019; 1-14
2045-2322
CONICET Digital
CONICET
url http://hdl.handle.net/11336/117400
identifier_str_mv Kosakyan, Anush; Alama Bermejo, Gema; Bartošová-Sojková, Pavla; Born-Torrijos, Ana; Šíma, Radek; et al.; Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites; Nature Publishing Group; Scientific Reports; 9; 1; 12-2019; 1-14
2045-2322
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-019-51479-0
info:eu-repo/semantics/altIdentifier/url/http://www.nature.com/articles/s41598-019-51479-0
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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