Reference Gene Selection for Gene Expression Analysis of Oocytes Collected from Dairy Cattle and Buffaloes during Winter and Summer

dc.contributorUniversidade Federal de São Carlos (UFSCar)
dc.contributorUniversidade de São Paulo (USP)
dc.contributorUniversidade Estadual Paulista (Unesp)
dc.contributorAgência Paulista de Tecnologia dos Agronegócios (APTA)
dc.contributorVida Reprod Consultoria
dc.contributorMSD Saude Anim
dc.contributorLTDA
dc.contributorUniv Montreal
dc.creatorMacabelli, Carolina Habermann
dc.creatorFerreira, Roberta Machado
dc.creatorGimenes, Lindsay Unno [UNESP]
dc.creatorTonizza de Carvalho, Nelcio Antonio
dc.creatorSoares, Julia Gleyci
dc.creatorAyres, Henderson
dc.creatorFerraz, Marcio Leao
dc.creatorWatanabe, Yeda Fumie
dc.creatorWatanabe, Osnir Yoshime
dc.creatorSangalli, Juliano Rodrigues
dc.creatorSmith, Lawrence Charles
dc.creatorBaruselli, Pietro Sampaio
dc.creatorMeirelles, Flavio Vieira
dc.creatorChiaratti, Marcos Roberto
dc.date2014-12-03T13:11:24Z
dc.date2014-12-03T13:11:24Z
dc.date2014-03-27
dc.date.accessioned2023-09-09T10:09:51Z
dc.date.available2023-09-09T10:09:51Z
dc.identifierhttp://dx.doi.org/10.1371/journal.pone.0093287
dc.identifierPlos One. San Francisco: Public Library Science, v. 9, n. 3, 13 p., 2014.
dc.identifier1932-6203
dc.identifierhttp://hdl.handle.net/11449/113078
dc.identifier10.1371/journal.pone.0093287
dc.identifierWOS:000333677500103
dc.identifierWOS000333677500103.pdf
dc.identifier9472065929531215
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8762691
dc.descriptionOocytes from dairy cattle and buffaloes have severely compromised developmental competence during summer. While analysis of gene expression is a powerful technique for understanding the factors affecting developmental hindrance in oocytes, analysis by real-time reverse transcription PCR (RT-PCR) relies on the correct normalization by reference genes showing stable expression. Furthermore, several studies have found that genes commonly used as reference standards do not behave as expected depending on cell type and experimental design. Hence, it is recommended to evaluate expression stability of candidate reference genes for a specific experimental condition before employing them as internal controls. In acknowledgment of the importance of seasonal effects on oocyte gene expression, the aim of this study was to evaluate the stability of expression levels of ten well-known reference genes (ACTB, GAPDH, GUSB, HIST1H2AG, HPRT1, PPIA, RPL15, SDHA, TBP and YWHAZ) using oocytes collected from different categories of dairy cattle and buffaloes during winter and summer. A normalization factor was provided for cattle (RPL15, PPIA and GUSB) and buffaloes (YWHAZ, GUSB and GAPDH) based on the expression of the three most stable reference genes in each species. Normalization of non-reference target genes by these reference genes was shown to be considerably different from normalization by less stable reference genes, further highlighting the need for careful selection of internal controls. Therefore, due to the high variability of reference genes among experimental groups, we conclude that data normalized by internal controls can be misleading and should be compared to not normalized data or to data normalized by an external control in order to better interpret the biological relevance of gene expression analysis.
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.descriptionCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.descriptionUniv Fed Sao Carlos, Ctr Ciencias Biol & Saude, Dept Genet & Evolucao, BR-13560 Sao Carlos, SP, Brazil
dc.descriptionUniv Sao Paulo, Fac Zootecnia & Engn Alimentos, Dept Med Vet, Pirassununga, SP, Brazil
dc.descriptionUniv Sao Paulo, Fac Med Vet & Zootecnia, Dept Reprod Anim, Sao Paulo, Brazil
dc.descriptionUniv Estadual Paulista, Fac Ciencias Agr & Vet, Dept Reprod Anim, Jaboticabal, SP, Brazil
dc.descriptionAPTA, Unidade Pesquisa Desenvolvimento Registro, Registro, SP, Brazil
dc.descriptionVida Reprod Consultoria, Cravinhos, SP, Brazil
dc.descriptionMSD Saude Anim, Sao Paulo, Brazil
dc.descriptionLTDA, WTA Tecnol Aplicada, Cravinhos, SP, Brazil
dc.descriptionUniv Sao Paulo, Fac Med Vet & Zootecnia, Dept Cirurgia, Sao Paulo, Brazil
dc.descriptionUniv Montreal, Fac Med Vet, Ctr Rech Reprod Anim, St Hyacinthe, PQ J2S 7C6, Canada
dc.descriptionUniv Estadual Paulista, Fac Ciencias Agr & Vet, Dept Reprod Anim, Jaboticabal, SP, Brazil
dc.descriptionFAPESP: 09/00938-3
dc.descriptionFAPESP: 10/13384-3
dc.descriptionFAPESP: 10/09561-7
dc.descriptionFAPESP: 11/14207-0
dc.descriptionFAPESP: 12/07510-1
dc.descriptionCNPq: 476229/2011-1
dc.format13
dc.languageeng
dc.publisherPublic Library Science
dc.relationPLOS ONE
dc.relation2.766
dc.relation1,164
dc.rightsAcesso aberto
dc.sourceWeb of Science
dc.titleReference Gene Selection for Gene Expression Analysis of Oocytes Collected from Dairy Cattle and Buffaloes during Winter and Summer
dc.typeArtigo


Este ítem pertenece a la siguiente institución