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Registro Completo |
Biblioteca(s): |
Embrapa Pecuária Sudeste. |
Data corrente: |
28/08/2018 |
Data da última atualização: |
12/09/2018 |
Tipo da produção científica: |
Artigo em Periódico Indexado |
Autoria: |
TOSCANO, J. H. B.; LOPES, L. G.; GIRALDELO, L. A.; SILVA, M. H. da; OKINO, C. H.; CHAGAS, A. C. de S. |
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. |
Título: |
Identification of appropriate reference genes for local immune-related studies in Morada Nova sheep infected with Haemonchus contortus. |
Ano de publicação: |
2018 |
Fonte/Imprenta: |
Molecular Biology Reports, v.45, n.5, p.1253-1262, out. 2018. |
Idioma: |
Inglês |
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 |
Palavras-Chave: |
Genes de referência. |
Thesagro: |
Haemonchus Contortus; Ovino. |
Thesaurus Nal: |
Sheep. |
Categoria do assunto: |
G Melhoramento Genético |
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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Biblioteca(s): |
Embrapa Semiárido. |
Data corrente: |
29/09/1999 |
Data da última atualização: |
15/07/2021 |
Tipo da produção científica: |
Resumo em Anais de Congresso |
Autoria: |
TAVARES, S. C. C. de H.; AMORIM, L. R. |
Afiliação: |
SELMA CAVALCANTI CRUZ DE H TAVARES, CPATSA. |
Título: |
Levantamento de Botryodiplodia theobromae em aréas irrigadas do trópico semi-árido brasileiro. |
Ano de publicação: |
1995 |
Fonte/Imprenta: |
Fitopatologia Brasileira, Brasília, DF, v. 20, p. 326, ago. 1995. |
Idioma: |
Português |
Notas: |
Suplemento. |
Conteúdo: |
Hoje, este fungo apresenta-se com severidade crescente infectando várias culturas potenciais. |
Palavras-Chave: |
Area irrigada; Diseases; Irrigated area; Plant; Regiao semi-arida; Semi-arid region. |
Thesagro: |
Botryodiplodia Theobromae; Doença; Planta. |
Categoria do assunto: |
H Saúde e Patologia |
URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/181129/1/Fitopatopatologia-Brasileira-v.20-p.326-1995.pdf
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Marc: |
LEADER 00798nam a2200241 a 4500 001 1133526 005 2021-07-15 008 1995 bl uuuu u00u1 u #d 100 1 $aTAVARES, S. C. C. de H. 245 $aLevantamento de Botryodiplodia theobromae em aréas irrigadas do trópico semi-árido brasileiro. 260 $aFitopatologia Brasileira, Brasília, DF, v. 20, p. 326, ago. 1995.$c1995 500 $aSuplemento. 520 $aHoje, este fungo apresenta-se com severidade crescente infectando várias culturas potenciais. 650 $aBotryodiplodia Theobromae 650 $aDoença 650 $aPlanta 653 $aArea irrigada 653 $aDiseases 653 $aIrrigated area 653 $aPlant 653 $aRegiao semi-arida 653 $aSemi-arid region 700 1 $aAMORIM, L. R.
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