Identification and expression analysis of genes associated with bovine blastocyst formation

Goossens, Karen; Soom, Ann Van; Poucke, Mario Van; Vandaele, Leen; Vandesompele, Jo; Zeveren, Alex Van; Peelman, Luc

doi:10.1186/1471-213x-7-64

Cited by 60 publications

(54 citation statements)

References 72 publications

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“…Elevated levels of the ATP5A1 transcript have been noted in bovine BCB -oocytes (Torner et al, 2008), which was related to increased metabolic activity of growing gametes. The EEF1A1 gene regulates protein synthesis (Moldave, 1985) and stimulates blastocyst development in cattle (Goossens et al, 2007). In vitro-produced bovine blastocysts contained significantly less transcripts of this gene when compared to their in vivo-derived counterparts, which usually display better quality.…”

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confidence: 99%

Maternal nutrition affects the composition of follicular fluid and transcript content in gilt oocytes

Warzych¹,

Cieślak²,

Pawlak³

et al. 2011

Vet. Med.

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Metabolomics focused on reproduction have been the subject of special interest in the past decade. Metabolite profiling provides detailed information on the status of follicular fluid and follicular cells which accompany the growing oocyte. Although nutrients present in the diet reach oocytes via the follicular fluid, it is not evident whether oocyte/embryo quality can be predicted based on the follicular fluid composition. Since this phenomenon has not yet been investigated in the pig, the aim of the present study was to investigate associations between diet composition and (1) selected markers related to developmental potential of oocytes (brilliant cresyl blue -BCB test, relative transcript abundance of EEF1A1 and ATP5A1 marker genes) and (2) fatty acid profile in the follicular fluid. Gilts were fed control and experimental diets which differed in composition and fatty acid profiles. The experimental diet used in this study comprised mainly locally grown grains (barley and rye) traditionally used for fattening pigs in Poland. Cumulus-oocyte complexes were aspirated from individual pairs of ovaries after animal slaughter, evaluated morphologically and subjected to the BCB test. Relative transcript abundance for the two marker genes was monitored by real-time PCR in oocytes of both categories (BCB + and BCB -). Fatty acid profile in follicular fluid was analyzed by gas chromatography. We show that the experimental diet rich in n-3 fatty acid significantly influences fatty acid composition of the follicular fluid. The fatty acid profile of the follicular fluid of gilts fed the experimental diet differed from that of the control females. The content of saturated fatty acids was higher in the experimental group, whereas unsaturated and polyunsaturated fatty acids were more abundant in the control group (P < 0.05). With regard to individual fatty acids, only C16:0 (palmitic acid), C18:2n-6 (linoleic acid) and C22:6 (docosahexaenoic acid) differed significantly. The abundance of ATP5A1 mRNA was influenced neither by diet composition nor by oocyte category (BCB + /BCB -) whereas that of the EEF1A1 was affected by both factors. Since higher mRNA level of the EEF1A1 gene was noted in BCB + oocytes this may be considered as a marker of oocyte quality in the pig. Maternal nutrition is one of the main factors affecting developmental competence of mammalian oocytes and embryos. Diet composition may alter the maternal environment of the oocyte and embryo growth through metabolic and endocrine modifications. Zak et al. (1997) showed that feed intake influenced the meiotic competence of porcine oocytes. They claimed that maternal under-nutrition may reduce the follicle's ability to support maturation of oocytes and thus their quality. Follicular fluid (FF) as an endogenous medium participates in transmitting nutrition signals to the

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mentioning

confidence: 99%

Maternal nutrition affects the composition of follicular fluid and transcript content in gilt oocytes

Warzych¹,

Cieślak²,

Pawlak³

et al. 2011

Vet. Med.

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“…These results are consistent with previous reports on the association of MYL6 with blastocyst formation. Goossens et al (2007) reported that MYL6 is highly expressed at the blastocyst stage compared with earlier stages of development and concluded that this gene could be a candidate marker for blastocyst formation. However, no data has been reported on the involvement of the different splice variants of MYL6 in early embryonic development.…”

Section: Resultsmentioning

confidence: 99%

Novel transcripts and alternatively spliced genes are associated with early development in bovine embryos

Zhang

Peñagaricano

Chen

et al. 2012

Animal

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Infertility in cattle is a major concern of farmers worldwide and despite the enormous improvements in assisted reproduction technologies, the success rates of pregnancies are still low. Embryonic loss is considered one of the main factors of infertility in cattle. As such, the identification of genetic markers for embryo quality and development can help elucidate the molecular mechanisms involved in the formation of embryos with the highest developmental potential. In a previous study, using nextgeneration RNA sequencing, we identified novel transcripts and alternatively spliced genes that were associated with embryo quality. The objectives of this study were to characterize these transcripts and validate their expression in new biological replications of embryos using quantitative real-time PCR. Two types of embryos differing in morphological and developmental statuses (blastocysts and degenerate embryos) were produced using in vitro fertilization. Quantitative expression of eight novel transcripts revealed a range of 2.5-to 90-fold difference in expression between degenerate embryos and blastocysts. Some of these novel transcripts showed sequence similarity to human and cattle genes known to affect differentiation, growth and development. In addition, expression analysis of alternative splicing isoforms of five genes (MYL6, NOP10, RNF187, RPS24 and RPS28) revealed significant differential expression of these isoforms in the different embryo types. Thus, results of this study suggest that novel transcripts and alternatively spliced genes, found to be differentially expressed between blastocysts and degenerate embryos, can be used as markers for blastocyst formation and development.Keywords: embryo, fertility, gene expression, novel transcripts, alternative splicing ImplicationsOver the last 40 to 50 years there has been a significant decline in reproductive performance of dairy cattle, due in part to low fertilization rates and early embryonic loss. Although genetics plays an important role in embryo development, the identification of genes associated with embryo quality and pregnancy success has been a serious challenge. In this study, we characterized several genes that showed altered expression in preimplantation embryos that do not develop normally to the blastocyst stage. These genes can be used as candidate markers for embryo quality in cattle. IntroductionIn the last few decades, production of embryos by in vitro fertilization (IVF) has become a common practice for the treatment of infertility in both humans and cattle, and although IVF procedures are considered to be a true revolution in the field of fertility, the success rate of IVF pregnancies is still low. For example, in humans, , 8 out of 10 transferred embryos fail to implant successfully (Bromer and Seli, 2008), most likely because of the inability to identify embryos with the best development potential. In cattle, there has been a significant decline in fertility over the past few decades (Dobson et al., 2007;Leroy et al., 2008) because of low...

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“…Na descrição do método, foi obtido apenas 3 a 5 vezes de multiplicação em uma única etapa, mas com modificações, esta taxa passou para 15 vezes a até 2.500 vezes em uma única etapa (BAUGH et al, 2001;DONNISON;PFEFFER, 2004;EBERWINE, 1996;GINSBERG et al, 2005;GOMES et al, 2003;JENSON et al, 2003;MOLL;DUSCHL;RICHTER, 2004;PHILLIPPS;EBERWINE, 1996;SCHNEIDER et al, 2004; VAN GELDER et al, 1990;WANG E. et al, 2000;WANG, 2005;ZHU;BABA, 2006 (BALASUBRAMANIAN et al, 2007;BUSTIN, 2002;GOOSSENS et al, 2007GOOSSENS et al, , 2005MISIRLIOGLU et al 2006;ROBERT et al, 2002;VIGNEAULT et al, 2007).…”

Section: Metodologias De Estudo Na Embriogêneseunclassified

“…Para fins de cálculo da regulação relativa, o GAPDH foi utilizado como gene de referência por apresentar a melhor eficiência de reação e menor variação de regulação que a H2A.z e o SDHA. Apesar de alguns autores não acreditarem que este seja um bom controle endógeno para estudos em embriões no estádio de pré-implantação por não manter os níveis de regulação constante entre os diferentes estádios (BUSTIN, 2002;ROBERT et al, 2002), trabalhos recentes utilizam o GAPDH como controle endógeno (BALASUBRAMANIAN et al, 2007;GOOSSENS et al, 2007;MISIRLIOGLU et al 2006).…”

Section: Função Molecularunclassified

“…Alguns autores não utilizam o GAPDH pela inconstância de expressão do mesmo em estudos pré-implantação (BUSTIN, 2002;ROBERT et al, 2002), enquanto outros acreditam que o GAPDH é um bom controle endógeno (BALASUBRAMANIAN et al, 2007;MISIRLIOGLU et al 2006). Há uma corrente que sugere a utilização de uma média geométrica entre três endógenos (GOOSSENS et al, 2007;GOOSSENS et al, 2005) ou utilização da H2A (JEONG et al, 2005;ROBERT et al, 2002;VIGNEAULT et al, 2007). No presente estudo, foram utilizados o GAPDH, a H2A.z e o SDHA.…”

Section: Análise Estatísticaunclassified

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Ativação de genes apoptóticos no bloqueio do desenvolvimento em embriões bovinos

Cortezzi¹

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Ao Léo, que entre muitas idas e vindas ao frigorífico, trazia não só os ovários, mas também seu sorriso e alto astral todos os dias. À Giovana, sempre muito solícita em ajudar em todas as etapas, incluindo nesta fase final devido à minha ausência. Obrigada pelo auxílio com os documentos! 5 A todos que fazem ou já fizeram parte do LMMD. São tantas pessoas ao longo destes anos que seria injusto citar nomes. Obrigada a todos por me ensinar que diferenças entre as pessoas fazem parte da vida, auxiliar com aspirações de ovários, ensinar procedimentos, discutir idéias e problemas, dividir churrascos e festas, e também auxiliar à distância sempre que necessário! Em especial ao Marcos e ao Moysés, fundamentais em algumas discussões deste trabalho.Às amigas da FZEA Claudinha e Fran, pela amizade e companhia nas baladas pirassununguenses, e à Estelinha, pela amizade e estadia nos momentos finais.Ao pessoal da dança, Marcos Mendonça, Marcos, Ursiley e Silvana, pela amizade e descontração não só nas aulas, mas principalmente fora delas. Com vocês eu pude relaxar entre boleros, sambas, tangos, salsas, forrós... Mesmo agora que estamos cada um em uma cidade, vocês sempre serão minha "turma de dança"! Aos meus amigos de Jaboticabal, antigos ou adquiridos pós-Ronaldo, pelos poucos mas animados finais de semana na terra natal. Em especial à Nádia, Dani (estadias e apoio em Sampa), Raqs Lima, Raqs Costa, Clau Dorigan, Márcia e família.Aos meus amigos da fase de São José do Rio Preto. Entre os muitos, destaco a Lê Vidotto (e seus pais) e a Lú Bizari, sempre disponibilizando suas casas para minha estadia. À Cássia e seus pais pelo carinho e amizade. Ao Eduardão, ao Adriano, ao Alfredo e ao Amílcar, que ficam na torcida de longe, via internet, me dão os puxões de orelha necessários e fazem minha vida mais feliz.Ao pessoal da Associação Instituto Sapientiae -Clínica Fertility, por acreditarem na minha capacidade profissional. analyze gene expression differences between fast cleavage and slow cleavage embryos, and discover possible signaling pathways to cell death or cell survival. We used a macroarray membrane spotted with human genes related to cell cycle and hybridizated with a radioactive labeled aRNA from fast or slow in vitro produced embryos. Real-time PCR and signaling pathways analysis were designed for further validation of the array. The mean similarity between human and bovine genes was 89,3%. According to the array membranes, it was possible to identify 120 genes differentially expressed between slow and fast cleavage embryos. Hence, the majority of the genes were more expressed in slow embryos (100 genes versus 20 genes in fast group). Among genes more expressed at fast embryos, 40%were identified as protein binding and 25% have catalytic activity, with similar results in slow embryos. In one hand, differences between fast and slow embryos transcripts were not confirmed by real-time PCR analysis. But on the other hand, the differentially expressed genes are somehow related to and constitutively present in some recognized...

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Identification and expression analysis of genes associated with bovine blastocyst formation

Abstract: Background: Normal preimplantation embryo development encompasses a series of events including first cleavage division, activation of the embryonic genome, compaction and blastocyst formation.

Cited by 60 publications

References 72 publications

Maternal nutrition affects the composition of follicular fluid and transcript content in gilt oocytes

Maternal nutrition affects the composition of follicular fluid and transcript content in gilt oocytes

Novel transcripts and alternatively spliced genes are associated with early development in bovine embryos

Ativação de genes apoptóticos no bloqueio do desenvolvimento em embriões bovinos

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