The oxidative stress adaptor p66Shc is required for permanent embryo arrest in vitro

Favetta, Laura A.; Madan, Pavneesh; Mastromonaco, Gabriela F.; John, Elizabeth J. St.; King, W.A.; Betts, Dean H.

doi:10.1186/1471-213x-7-132

Cited by 52 publications

(52 citation statements)

References 77 publications

(99 reference statements)

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“…shc is up-regulated in the early arrest, while p53 does not seem to be involved in the same function (Favetta et al 2004(Favetta et al , 2007a(Favetta et al , 2007b. Apoptosis is involved in the bovine early developmental arrest in vivo (Yang & Rajamahendran 2002), although developmental arrest independent of apoptosis has been also described to occur in the uterus .…”

Section: Development and Gene Expression In Parthenotesmentioning

confidence: 97%

Biological differences between in vitro produced bovine embryos and parthenotes

Gómez¹,

Gutiérrez‐Adán²,

Díez³

et al. 2009

Reproduction

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Parthenotes may represent an alternate ethical source of stem cells, once biological differences between parthenotes and embryos can be understood. In this study, we analyzed development, trophectoderm (TE) differentiation, apoptosis/necrosis, and ploidy in parthenotes and in vitro produced bovine embryos. Subsequently, using real-time PCR, we analyzed the expression of genes expected to underlie the observed differences at the blastocyst stage. In vitro matured oocytes were either fertilized or activated with ionomycin C6-DMAP and cultured in simple medium. Parthenotes showed enhanced blastocyst development and diploidy and reduced TE cell counts. Apoptotic and necrotic indexes did not vary, but parthenotes evidenced a higher relative proportion of apoptotic cells between inner cell mass and TE. The pluripotence-related POU5F1 and the methylation DNMT3A genes were downregulated in parthenotes. Among pregnancy recognition genes, TP-1 was upregulated in parthenotes, while PGRMC1 and PLAC8 did not change. Expression of p66 shc and BAX/BCL2 ratio were higher, and p53 lower, in parthenotes. Among metabolism genes, SLC2A1 was downregulated, while AKR1B1, PTGS2, H6PD, and TXN were upregulated in parthenotes, and SLC2A5 did not differ. Among genes involved in compaction/blastulation, GJA1 was downregulated in parthenotes, but no differences were detected within ATP1A1 and CDH1. Within parthenotes, the expression levels of SLC2A1, TP-1, and H6PD, and possibly AKR1B1, resemble patterns described in female embryos. The pro-apoptotic profile is more pronounced in parthenotes than in embryos, which may differ in their way to channel apoptotic stimuli, through p66 shc and p53respectively, and in their mechanisms to control pluripotency and de novo methylation.Reproduction (2009) 137 285-295

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Section: Development and Gene Expression In Parthenotesmentioning

confidence: 97%

Biological differences between in vitro produced bovine embryos and parthenotes

Gómez¹,

Gutiérrez‐Adán²,

Díez³

et al. 2009

Reproduction

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“…After TCN counting, the fluorescence intensity of individual embryos was assessed in one central optical section displaying both the ICM and the TE by marking and extracting six small areas of equal size. Extracted images were saved in tagged image file format (TIFF) and processed according to the method developed by Tolivia et al (2006) and validated in bovine embryos (Favetta et al 2007). Briefly, green channel images (representing Alexa Fluor 488 fluorescence) were converted to 'greyscale' and then inverted with Adobe Photoshop CS2 (Adobe Systems Inc.) so that grey and black pixels represented the areas of Alexa Fluor 488 immunofluorescence on a white background.…”

Section: Immunofluorescencementioning

confidence: 99%

Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor

Velazquez¹,

Hermann²,

Kues³

et al. 2011

Reproduction

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The hypothesis that high concentrations of IGF1 can impair embryo development was investigated in a bovine in vitro model to reflect conditions in polycystic ovary syndrome (PCOS) patients. Embryos were either cultured in the absence or presence of a physiological (100 ng/ml) or supraphysiological (1000 ng/ml) IGF1 concentration. Cell allocation, apoptosis, transcript and protein expression of selected genes involved in apoptosis, glucose metabolism and the IGF system were analysed. Supraphysiological IGF1 concentration did not improve blastocyst formation over controls, but induced higher levels of apoptosis, decreased TP53 protein expression in the trophectoderm and increased the number of cells in the inner cell mass (ICM). The increase in ICM cells corresponded with an increase in IGF1 receptor (IGF1R) protein in the ICM. A small, but significant, percentage of blastocysts displayed a hypertrophic ICM, not observed in controls and virtually absent in embryos treated with physiological concentrations of IGF1. Physiological IGF1 concentrations increased total IGF1R protein expression and upregulated IGFBP3 transcripts leading to an increase in blastocyst formation with no effects on cell number or apoptosis. In conclusion, the results support the hypothesis of detrimental effects of supraphysiological IGF1 concentrations on early pregnancy. However, our results do not support the premise that increased apoptosis associated with high levels of IGF1 is mediated via downregulation of the IGF1R as previously found in preimplantation mouse embryos. This in vitro system with the bovine preimplantation embryo reflects critical features of fertility in PCOS patients and could thus serve as a useful model for in-depth mechanistic studies.

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“…Realmente, foi detectada uma maior proporção de transcritos em embriões com alta taxa de bloqueio do que naqueles embriões ditos competentes ao desenvolvimento (DODE et al, 2006;FAVETTA et al, 2007;FIGUEIREDO, 2005;JEONG et al, 2006;MATWEE;KING, 2000;MEIRELLES et al, 2004). Estudos observaram a ocorrência de MCP de forma espontânea apenas após a AGE, ou que com o uso de indutores químicos, a apoptose ocorreu em maior grau após a AGE, contrariando que a MCP seria um processo passivo por ausência de transcrição GJORRET et al, 2007).…”

Section: Introductionunclassified

“…Vários estudos verificaram a expressão de um grande número de genes em embriões com alta taxa de bloqueio quando comparados a embriões com baixa taxa de bloqueio, incluindo genes importantes cuja transcrição diferencial está associada à sobrevivência ou morte celular (DODE et al, 2006;FAVETTA et al, 2007;FIGUEIREDO, 2005;JEONG et al, 2006;MATWEE;KING, 2000;MEIRELLES et al, 2004). Observações em várias espécies, incluindo a bovina, mostram que a MCP espontânea não ocorre em estádios anteriores a AGE, e a resposta a indutores de MCP no meio de cultivo também é baixa antes da AGE.…”

Section: Introductionunclassified

“…Como já descrito, vários estudos verificaram uma maior expressão de genes em embriões com alta taxa de bloqueio quando comparados a embriões com baixa taxa de bloqueio, incluindo genes vinculados a vias de sinalização de MCP (DODE et al, 2006;FAVETTA et al, 2007;FIGUEIREDO, 2005;JEONG et al, 2006;MATWEE;KING, 2000;MEIRELLES et al, 2004). Devido à ocorrência de apoptose espontânea apenas após a AGE em embriões bovinos, este bloqueio do desenvolvimento parece ser um mecanismo ativo, baseado no balanceamento entre proteínas pró e anti-apoptóticas, mas não envolvendo os membros da família Bcl-2 (BADR et al, 2007;FABIAN et al, 2007;GJORRET et al, 2003;JURISICOVA et al, 1998;MATWEE;KING, 2000;MEIRELLES et al, 2004;OPIELA et al, 2008;YANG & RAJAMAHENDRAN, 2002).…”

Section: Introductionunclassified

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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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The oxidative stress adaptor p66Shc is required for permanent embryo arrest in vitro

Cited by 52 publications

References 77 publications

Biological differences between in vitro produced bovine embryos and parthenotes

Biological differences between in vitro produced bovine embryos and parthenotes

Increased apoptosis in bovine blastocysts exposed to high levels of IGF1 is not associated with downregulation of the IGF1 receptor

Ativação de genes apoptóticos no bloqueio do desenvolvimento em embriões bovinos

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