Culture of Zygotes Increases p53 Expression in B6 Mouse Embryos, which Reduces Embryo Viability1

Li, A.; Chandrakanthan, Vashe; Chami, Omar; O’Neill, Chris

doi:10.1095/biolreprod.106.056838

Cited by 48 publications

(60 citation statements)

References 30 publications

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“…Our results show that culture in KSOM did not reduce the number of cells at the blastocyst stage; however, we have identified a number of cellular stressors induced by the culture environment (KSOM with or without FCS), including apoptosis and oxidative stressrelated genes (Trp53inp1, Zmat3, Pdrg1, Cdkn1a, Prdx2). In agreement with our results, it has been reported that the culture of mouse zygotes increases Trp53 expression and reduces embryo viability, suggesting that monitoring Trp53 expression in embryos may be a tool for assessing the effects of IVC techniques on embryo viability (Li et al 2007). The expression of genes related to membrane transport function was also altered by IVC (Prdx2, Slc7a3, Slc15a2) in agreement with previous reports (Rinaudo & Schultz 2004).…”

Section: Discussionsupporting

confidence: 93%

Analysis of gene transcription alterations at the blastocyst stage related to the long-term consequences of in vitro culture in mice

Fernández‐González¹,

Hourcade²,

López‐Vidriero³

et al. 2009

Reproduction

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We have reported that in vitro culture (IVC) of preimplantation mouse embryos in the presence of FCS produces long-term effects (LTE) on development, growth and behaviour of the offspring at adult age. To analyse the mechanisms underlying this phenomenon, we have examined development and global alterations in gene expression in the mouse blastocysts produced in the presence of FCS, conditions known to be suboptimal and that generate LTE. Embryos cultured in vitro in KSOM and in KSOMCFCS had a reduced number of cells in the inner cell mass at the blastocyst stage compared with in vivo derived embryos; however, only culture in KSOMCFCS leads to a reduction in the number of trophoblast cells. Gene expression levels were measured by comparison among three groups of blastocysts (in vivo, IVC in KSOM and IVC in KSOMCFCS). Different patterns of gene expression and development were found between embryos cultured in vitro or in vivo. Moreover, when we compared the embryos produced in KSOM versus KSOMCFCS, we observed that the presence of FCS affected the expression of 198 genes. Metabolism, proliferation, apoptosis and morphogenetic pathways were the most common processes affected by IVC. However, the presence of FCS during IVC preferentially affected genes associated with certain molecular and biological functions related to epigenetic mechanisms. These results suggest that culture-induced alterations in transcription at the blastocyst stage related to epigenetic mechanisms provide a foundation for understanding the molecular origin at the time of preimplantation development of the long-term consequences of IVC in mammals.

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Section: Discussionsupporting

confidence: 93%

Analysis of gene transcription alterations at the blastocyst stage related to the long-term consequences of in vitro culture in mice

Fernández‐González¹,

Hourcade²,

López‐Vidriero³

et al. 2009

Reproduction

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“…However, a 24 h exposure does not reduce the development rate [9], unlike that which occurs with a 48 h overexposure. A similar reduction in development rates, in coincidence with a strong up-regulation and nuclear accumulation of p53, has been reported in mouse blastocysts cultured in vitro, but not in their in vivo counterparts [34]. In addition, bax expression correlated with p53 expression in ATRA and untreated controls, while this association was not observed under LG stimulation; this is consistent with the detected responsiveness of p53 to ATRA but not to LG.…”

Section: Discussionsupporting

confidence: 87%

Retinoid receptor-specific agonists regulate bovine in vitro early embryonic development, differentiation and expression of genes related to cell cycle arrest and apoptosis

et al. 2007

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A major goal in reproductive biotechnology is the identification of pathways that regulate early embryonic development and the allocation of cells to the inner cell mass (ICM) and trophectoderm (TE). Retinoids regulate the development and differentiation of the bovine blastocyst in vitro, although the involvement of the retinoid X receptors (RXRs) remains to be clarified. This paper compares the effect of a synthetic RXR agonist (LG100268; LG) with that of the retinoic acid receptor (RAR) agonist all-trans retinoic acid (ATRA) on blastulation. In vitro-produced morulae were treated for 48 h with LG (0.1 mM, 1 mM and 10 mM), ATRA 0.7 mM, or no additives. Treatment with ATRA did not increase the rate of development; however, the LG 0.1 mM treatment increased both the blastocyst development and hatching rate. Cell numbers increased in the ICM with LG 10 mM, while a dosedependent reduction was observed in the TE in the presence of LG. Gene expression levels of p53 and p66 did not vary with LG but increased with ATRA. Both LG and ATRA activated bax, a pro-apoptotic gene and H2A.Z, a cell cycle-related gene. The above effects suggest the existence of active p53-dependent and -independent apoptotic pathways for ATRA and LG, respectively, in the bovine embryo. The expression of p53 and H2A.Z showed a strong, positive correlation (r = 0.93; p < 0.0001) in all experimental groups; both proteins are linked through the cell cycle. Agonists of RXR could be used to control blastocyst development and differentiation. #

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“…Specifically, active p53 has been shown to cause differentiation in ESCs by binding to the Nanog promoter region and repressing expression [35]. Additionally, the in vitro culture of embryos has been shown to increase p53 activity, with p53-dependant negative effects on embryo development and viability [83], while culture with pifithrina, an inhibitor of p53, has been shown to improve the proportion of embryos that develop to the blastocyst stage [38]. When p53 was indirectly activated by inhibiting SIRT-1 [37] with nicotinamide, insulin's ability to increase epiblast cell number was, as with GSK3 activation and PI3K inhibition, ameliorated.…”

Section: Discussionmentioning

confidence: 99%

Insulin Increases Epiblast Cell Number of In Vitro Cultured Mouse Embryos via the PI3K/GSK3/p53 Pathway

Campbell

Nottle

Vassiliev

et al. 2012

Stem Cells and Development

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Authors may also deposit this version on his/her funder's or funder's designated repository at the funder's request or as a result of a legal obligation, provided it is not made publicly available until 12 months after official publication. th March 2013Insulin High-quality embryos give rise to embryonic stem cells (ESCs) at greater efficiencies than poor-quality embryos. However, most embryos available for human ESC derivation are of a reduced quality as a result of culture in relatively simple media up to 10 years earlier, before cryopreservation, or before compaction. In the present study, we used a mouse model to determine whether a culture with insulin from the 8-cell stage could increase the number of ESC progenitor epiblast cells in blastocysts, as well as endeavor to determine the molecular mechanism of the insulin's effect. Culture in media containing 1.7 rM insulin increased epiblast cell number (determined by Oct4 and Nanog co-expression), and proportion in day 6 blastocysts. The inhibition of phosphoinositide 3 kinase (PI3K) (via LY294002), an early second messenger of the insulin receptor, blocked this effect. The inhibition of glycogen synthase kinase 3 (GSK3) or p53, 2 s messengers inactivated by insulin signaling (via CT99021 or pifithrin-a, respectively), increased epiblast cell numbers. When active, GSK3 and p53 block the transcription of Nanog, which is important for maintaining pluripotency. A simultaneous inhibition of GSK3 and p53 had no synergistic effects on epiblast cell number. The induced activation of GSK3 and p53, via the inhibition of proteins responsible for their inactivation (PKA via H-89 and SIRT-1 via nicotinamide, respectively), blocked the insulin's effect on the epiblast.From our findings, we conclude that insulin increases epiblast cell number via the activation of PI3K, which ultimately inactivates GSK3 and p53. Furthermore, we suggest that the inclusion of insulin in culture media could be used as a strategy for increasing the efficiency with which the ESC lines can be derived from cultured embryos.

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Culture of Zygotes Increases p53 Expression in B6 Mouse Embryos, which Reduces Embryo Viability1

Cited by 48 publications

References 30 publications

Analysis of gene transcription alterations at the blastocyst stage related to the long-term consequences of in vitro culture in mice

Analysis of gene transcription alterations at the blastocyst stage related to the long-term consequences of in vitro culture in mice

Retinoid receptor-specific agonists regulate bovine in vitro early embryonic development, differentiation and expression of genes related to cell cycle arrest and apoptosis

Insulin Increases Epiblast Cell Number of In Vitro Cultured Mouse Embryos via the PI3K/GSK3/p53 Pathway

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