Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris

Autores
Odetti, Lucia Magdalena; Paravani, Enrique Valentin; Simoniello, Maria Fernanda; Poletta, Gisela Laura
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The quantitative real-time polymerase chain reaction (qPCR) has been one of the most promising approaches to perform rapid and accurate quantification of DNA in various biological systems. The aim of this study was to standardized the qPCR technique for the analysis of important genes involved in the main routes of antioxidant defense against reactive oxygen species (catalase: cat and superoxide dismutase: sod) and evaluate the stability of different reference genes in blood of Caiman latirostris hatchlings. The stability of the reference genes, β-actin, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and ribosomal protein L8 (rpl8) was determined using the comparative ΔCt, NormFinder, geNorm, BestKeeper and RefFinder. Then, cat and sod genes were normalized with each reference gene and their mRNA abundances were determined through the qPCR. Stability of genes was ranked through the different methods in the following order: β-actin, rpl8 and gapdh, under normal physiological conditions. The results reveal that cat and sod genes present a similar relative mRNA abundance with β-actin and rpl8. This is the first report of the analysis of antioxidant mRNA as potential biomarkers of oxidative stress in blood for all crocodilians species. Besides, we determined the stability of different reference genes that can be used for normalization of mRNA abundance patterns in blood of C. latirostris, without the need to sacrifice the animals.
Fil: Odetti, Lucia Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina
Fil: Paravani, Enrique Valentin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentina
Fil: Simoniello, Maria Fernanda. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina
Fil: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentina
Materia
ANTIOXIDANT GENES
CROCODILES
MRNA ABUNDANCE PATTERN
REFERENCE GENES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/167382
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/167382
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostrisOdetti, Lucia MagdalenaParavani, Enrique ValentinSimoniello, Maria FernandaPoletta, Gisela LauraANTIOXIDANT GENESCROCODILESMRNA ABUNDANCE PATTERNREFERENCE GENEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The quantitative real-time polymerase chain reaction (qPCR) has been one of the most promising approaches to perform rapid and accurate quantification of DNA in various biological systems. The aim of this study was to standardized the qPCR technique for the analysis of important genes involved in the main routes of antioxidant defense against reactive oxygen species (catalase: cat and superoxide dismutase: sod) and evaluate the stability of different reference genes in blood of Caiman latirostris hatchlings. The stability of the reference genes, β-actin, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and ribosomal protein L8 (rpl8) was determined using the comparative ΔCt, NormFinder, geNorm, BestKeeper and RefFinder. Then, cat and sod genes were normalized with each reference gene and their mRNA abundances were determined through the qPCR. Stability of genes was ranked through the different methods in the following order: β-actin, rpl8 and gapdh, under normal physiological conditions. The results reveal that cat and sod genes present a similar relative mRNA abundance with β-actin and rpl8. This is the first report of the analysis of antioxidant mRNA as potential biomarkers of oxidative stress in blood for all crocodilians species. Besides, we determined the stability of different reference genes that can be used for normalization of mRNA abundance patterns in blood of C. latirostris, without the need to sacrifice the animals.Fil: Odetti, Lucia Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; ArgentinaFil: Paravani, Enrique Valentin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; ArgentinaFil: Simoniello, Maria Fernanda. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; ArgentinaFil: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); ArgentinaElsevier Science2021-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/167382Odetti, Lucia Magdalena; Paravani, Enrique Valentin; Simoniello, Maria Fernanda; Poletta, Gisela Laura; Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris; Elsevier Science; Heliyon; 7; 2; 2-2021; 6253-62532405-8440CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S2405844021003583info:eu-repo/semantics/altIdentifier/doi/10.1016/j.heliyon.2021.e06253info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:28:58Zoai:ri.conicet.gov.ar:11336/167382instacron: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 10:28:58.871CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
title Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
spellingShingle Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
Odetti, Lucia Magdalena
ANTIOXIDANT GENES
CROCODILES
MRNA ABUNDANCE PATTERN
REFERENCE GENES
title_short Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
title_full Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
title_fullStr Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
title_full_unstemmed Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
title_sort Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris
dc.creator.none.fl_str_mv Odetti, Lucia Magdalena
Paravani, Enrique Valentin
Simoniello, Maria Fernanda
Poletta, Gisela Laura
author Odetti, Lucia Magdalena
author_facet Odetti, Lucia Magdalena
Paravani, Enrique Valentin
Simoniello, Maria Fernanda
Poletta, Gisela Laura
author_role author
author2 Paravani, Enrique Valentin
Simoniello, Maria Fernanda
Poletta, Gisela Laura
author2_role author
author
author
dc.subject.none.fl_str_mv ANTIOXIDANT GENES
CROCODILES
MRNA ABUNDANCE PATTERN
REFERENCE GENES
topic ANTIOXIDANT GENES
CROCODILES
MRNA ABUNDANCE PATTERN
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 The quantitative real-time polymerase chain reaction (qPCR) has been one of the most promising approaches to perform rapid and accurate quantification of DNA in various biological systems. The aim of this study was to standardized the qPCR technique for the analysis of important genes involved in the main routes of antioxidant defense against reactive oxygen species (catalase: cat and superoxide dismutase: sod) and evaluate the stability of different reference genes in blood of Caiman latirostris hatchlings. The stability of the reference genes, β-actin, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and ribosomal protein L8 (rpl8) was determined using the comparative ΔCt, NormFinder, geNorm, BestKeeper and RefFinder. Then, cat and sod genes were normalized with each reference gene and their mRNA abundances were determined through the qPCR. Stability of genes was ranked through the different methods in the following order: β-actin, rpl8 and gapdh, under normal physiological conditions. The results reveal that cat and sod genes present a similar relative mRNA abundance with β-actin and rpl8. This is the first report of the analysis of antioxidant mRNA as potential biomarkers of oxidative stress in blood for all crocodilians species. Besides, we determined the stability of different reference genes that can be used for normalization of mRNA abundance patterns in blood of C. latirostris, without the need to sacrifice the animals.
Fil: Odetti, Lucia Magdalena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina
Fil: Paravani, Enrique Valentin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentina
Fil: Simoniello, Maria Fernanda. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina
Fil: Poletta, Gisela Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Cátedra de Toxicología y Bioquímica Legal; Argentina. Universidad Nacional del Litoral. Facultad de Humanidades y Ciencias. Departamento de Ciencias Naturales. Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA); Argentina
description The quantitative real-time polymerase chain reaction (qPCR) has been one of the most promising approaches to perform rapid and accurate quantification of DNA in various biological systems. The aim of this study was to standardized the qPCR technique for the analysis of important genes involved in the main routes of antioxidant defense against reactive oxygen species (catalase: cat and superoxide dismutase: sod) and evaluate the stability of different reference genes in blood of Caiman latirostris hatchlings. The stability of the reference genes, β-actin, glyceraldehyde 3-phosphate dehydrogenase (gapdh) and ribosomal protein L8 (rpl8) was determined using the comparative ΔCt, NormFinder, geNorm, BestKeeper and RefFinder. Then, cat and sod genes were normalized with each reference gene and their mRNA abundances were determined through the qPCR. Stability of genes was ranked through the different methods in the following order: β-actin, rpl8 and gapdh, under normal physiological conditions. The results reveal that cat and sod genes present a similar relative mRNA abundance with β-actin and rpl8. This is the first report of the analysis of antioxidant mRNA as potential biomarkers of oxidative stress in blood for all crocodilians species. Besides, we determined the stability of different reference genes that can be used for normalization of mRNA abundance patterns in blood of C. latirostris, without the need to sacrifice the animals.
publishDate 2021
dc.date.none.fl_str_mv 2021-02
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/11336/167382
Odetti, Lucia Magdalena; Paravani, Enrique Valentin; Simoniello, Maria Fernanda; Poletta, Gisela Laura; Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris; Elsevier Science; Heliyon; 7; 2; 2-2021; 6253-6253
2405-8440
CONICET Digital
CONICET
url http://hdl.handle.net/11336/167382
identifier_str_mv Odetti, Lucia Magdalena; Paravani, Enrique Valentin; Simoniello, Maria Fernanda; Poletta, Gisela Laura; Identification and evaluation of antioxidant and reference genes for quantitative real-time PCR in blood of Caiman latirostris; Elsevier Science; Heliyon; 7; 2; 2-2021; 6253-6253
2405-8440
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S2405844021003583
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.heliyon.2021.e06253
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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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