Abstract
Precise normalization with reference genes is necessary, in order to obtain reliable relative expression data in response to gastrointestinal nematode infection. By using sheep from temperate regions as models, three reference genes, viz., ribosomal protein LO (RPLO), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase complex subunit A (SDHA), were investigated in the abomasum, abomasal lymph nodes and small intestine of Brazilian Somalis sheep, either resistant or susceptible to gastrointestinal nematodes infections. Real time PCR was carried out by using SYBR Green I dye, and gene stability was tested by geNorm. RPLO was an ideal reference gene, since its expression was constant across treatments, presented lower variation, and was ranked as the most stable in abomasum and lymph node tissues. On the other hand, SDHA was the most stable in the small intestine followed by RPLO and GAPDH. These findings demonstrate the importance of correctly choosing reference genes prior to relative quantification. In addition, we determined that reference genes used in sheep from temperate regions, when properly tested, can be applied in animals from tropical regions such as the Brazilian Somalis sheep.
Highlights
Short CommunicationEvaluation of reference genes for real-time PCR studies of Brazilian Somalis sheep infected by gastrointestinal nematodes
Gastrointestinal nematode parasitism causes severe losses in sheep production (Ingham et al, 2008). Animals respond to this infection by activating the proliferation of T Helper (TH) which secretes cytokines related to resistance and susceptibility to gastrointestinal nematode infections (Zaros et al, 2007)
The study of these and other molecules is important for understanding the mechanisms involved in immune response to gastrointestinal nematode infections
Summary
Evaluation of reference genes for real-time PCR studies of Brazilian Somalis sheep infected by gastrointestinal nematodes. Lilian Giotto Zaros, Luiz Lehmann Coutinho, Lúcia Helena Sider, Henrique Rocha de Medeiros, Maria Rosalba Moreira das Neves, Camila Loures Benvenuti, Andrine Maria do Carmo Navarro and Luiz da Silva Vieira3 1Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil. Lilian Giotto Zaros, Luiz Lehmann Coutinho, Lúcia Helena Sider, Henrique Rocha de Medeiros, Maria Rosalba Moreira das Neves, Camila Loures Benvenuti, Andrine Maria do Carmo Navarro and Luiz da Silva Vieira3 1Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil. 2Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP, Brazil. 3Embrapa Caprinos e Ovinos, Sobral, CE, Brazil. 4Universidade Estadual Vale do Acaraú, Sobral, CE, Brazil
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