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Registro Completo |
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Biblioteca(s): |
Embrapa Pecuária Sudeste. |
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Data corrente: |
28/08/2018 |
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Data da última atualização: |
12/09/2018 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
TOSCANO, J. H. B.; LOPES, L. G.; GIRALDELO, L. A.; SILVA, M. H. da; OKINO, C. H.; CHAGAS, A. C. de S. |
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Afiliação: |
João Henrique Barbosa Toscano, UNESP; Louyse Gabrielli Lopes, UNICEP; Luciana Aparecida Giraldelo, UNICEP; Matheus Henrique da Silva, UNICEP; CINTIA HIROMI OKINO, CPPSE; ANA CAROLINA DE SOUZA CHAGAS, CPPSE. |
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Título: |
Identification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus. |
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Ano de publicação: |
2018 |
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Fonte/Imprenta: |
Molecular Biology Reports, v.45, n.5, p.1253-1262, out. 2018. |
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Idioma: |
Inglês |
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Conteúdo: |
Due to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results. MenosDue to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes,... Mostrar Tudo |
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Palavras-Chave: |
Genes de referência. |
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Thesagro: |
Haemonchus Contortus; Ovino. |
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Thesaurus Nal: |
Sheep. |
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Categoria do assunto: |
G Melhoramento Genético |
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Marc: |
LEADER 02463naa a2200229 a 4500
001 2094611
005 2018-09-12
008 2018 bl uuuu u00u1 u #d
100 1 $aTOSCANO, J. H. B.
245 $aIdentification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus.$h[electronic resource]
260 $c2018
520 $aDue to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results.
650 $aSheep
650 $aHaemonchus Contortus
650 $aOvino
653 $aGenes de referência
700 1 $aLOPES, L. G.
700 1 $aGIRALDELO, L. A.
700 1 $aSILVA, M. H. da
700 1 $aOKINO, C. H.
700 1 $aCHAGAS, A. C. de S.
773 $tMolecular Biology Reports$gv.45, n.5, p.1253-1262, out. 2018.
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Embrapa Pecuária Sudeste (CPPSE) |
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| 6. |  | RIBEIRO, J. M.; MELO, N. F. de; COELHO, A. K. N. dos S.; PINTO, M. dos S. T. Efeito do melado de cana-de-açúcar no desenvolvimento in vitro de bananeira (Musa spp.) cv. Maçã. Revista Ceres, Viçosa, MG, v. 59, n. 3, p. 293-298, mai/jun, 2012.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 2 |
| Biblioteca(s): Embrapa Semiárido. |
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| 10. | | RIBEIRO, J. M.; TEIXEIRA, S. L.; TEIXEIRA, M. T.; PINTO, M. dos S. T.; BASTOS, D. C. Influência do intervalo de inoculação de explantes e do hipoclorito de sódio na formação de brotos em Pfaffia glomerata (Spreng.) Pedersen. In: CONGRESSO BRASILEIRO DE FLORICULTURA E PLANTAS ORNAMENTAIS, 18.; CONGRESSO BRASILEIRO DE CULTURA DE TECIDOS DE PLANTAS, 5., 2011, Joinville. Resumos... Joinville: Sociedade Brasileira de Plantas Ornamentais e Associação Brasileira de Cultura de Tecidos de Plantas, 2011.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Semiárido. |
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| 12. | | RIBEIRO, J. M.; TEIXEIRA, S. L.; TEIXEIRA, M. T.; PINTO, M. dos S. T.; BASTOS, D. C. Hipoclorito de sódio para esterilização química de meios nutritivos e frascos de cultivo. In: CONGRESSO BRASILEIRO DE FLORICULTURA E PLANTAS ORNAMENTAIS, 18.; CONGRESSO BRASILEIRO DE CULTURA DE TECIDOS DE PLANTAS, 5., 2011, Joinville. Resumos... Joinville: Sociedade Brasileira de Plantas Ornamentais e Associação Brasileira de Cultura de Tecidos de Plantas, 2011.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Semiárido. |
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| 14. | | RIBEIRO, J. M.; MELO, N. F. de; COELHO, A. K. N. dos S.; PINTO, M. dos S. T. Uso da rapadura como meio nutritivo para cultivo in vitro de bananeira (Musa spp.) cv maçã. In: CONGRESSO BRASILEIRO DE FLORICULTURA E PLANTAS ORNAMENTAIS, 19.; CONGRESSO BRASILEIRO DE CULTURA DE TECIDOS DE PLANTAS, 6., 2013, Recife. Inovação tecnológica: resgate histórico, desafios e perspectivas. Recife: UFRPE, 2013. 1 CD-ROM.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Semiárido. |
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| 17. | | RIBEIRO, J. M.; OLIVEIRA, E. A. G. de; ARAUJO, F. P. de; FERNANDES, K. V. S.; MELO, N. F. de; PINTO, M. dos S. T. Análise preliminar da composição protéica de exsudatos de tuberculos de umbu (Spondias tuberosa Arruda), sob estresse hídrico. In: CONGRESSO BRASILEIRO DE FRUTICULTURA, 21., 2010, Natal. Frutas: saúde, inovação e responsabilidade: anais. Natal: SBF, 2010. 1 CD-ROM.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Semiárido. |
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| 18. | | RIBEIRO, J. M.; CASTRO, J. M. da C. e; MELO, N. F. de; FERNANDES, K. V. S.; PINTO, M. dos S. T. Efeito de extratos proteicos de amendoim sobre o desenvolvimento, a capacidade infectiva e atividade de enzimas proteolíticas de Meloidogyne enterolobii. Revista Ceres, v. 61, n. 3, p. 439-442, mai/jun. 2014.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Semiárido. |
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| 19. | | RIBEIRO, J. M.; MELO, N. F. de; ARAUJO, F. P. de; COELHO, A. K. N. dos S.; COELHO, M. do S. E.; PINTO, M. dos S. T. Micropropagação de goiabeira, maracujazeiro, bananeira e videira. Petrolina: Embrapa Semiárido, 2012. 7 p. il. (Embrapa Semiárido. Circular técnica, 101).| Biblioteca(s): Embrapa Semiárido. |
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| 20. | | PINTO, M. dos S, T.; RIBEIRO, J. M.; ARAUJO, F. P. de; MAIA, A. M. da S.; MELO, N. F. de; FERNANDES, K. V. S. Isolation and characterization of a peroxidase from Spondias tuberosa and its potential role as biotechnological tool. In: ANNUAL MEETING OF THE BRAZILIAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY, 45., 2016, Natal. Abstracts... São Paulo: SBBq, 2016.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Semiárido. |
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| Registros recuperados : 25 | |
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