Transcriptome and epigenome analysis of porcine embryos from non-esterified fatty acid-exposed oocytes

Shi, Meihong; Sirard, Marc‐André

doi:10.1016/j.domaniend.2021.106605

Cited by 6 publications

(21 citation statements)

References 55 publications

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“…The actual low differences and weak experimental effects collectively contribute to the less significant RT-qPCR results. Different significance power of the genes expression among the treatments between microarray and RT-qPCR validation are also found in other studies (Kim et al, 2012;Pagé-Larivière et al, 2016;M. Shi & Sirard, 2021), especially in the genes that have low fold change between treatments (Feng et al, 2010).…”

Section: Analysis Of Dna Methylation Datamentioning

confidence: 56%

“…Tables 4 and 5 & Sirard, 2021). Therefore, the presence of granulosa cells during maturation seems to lessen the effect of elevated NEFAs.…”

Section: Analysis Of Dna Methylation Datamentioning

confidence: 89%

“…A bar of length 1 indicates that twice as many probes as expected were differentially methylated. IVM, in vitro maturation; NEFA, nonesterified fatty acid exposed to same concentrations of NEFA during IVM in the absence of granulosa cells (M. Shi & Sirard, 2021). These most affected canonical pathways and upstream regulators demonstrated that the cells in blastocysts were responding to apoptosis and inflammation initiated by the exposure to excessive NEFAs during IVM.…”

Section: Analysis Of Dna Methylation Datamentioning

confidence: 99%

“…Follicular NEFAs play an important role in oocyte and embryo developments, and their high concentrations may have crossgenerationally detrimental effects. In our previous study, altered transcriptome and DNA methylation patterns were found in porcine blastocysts derived from pathologically elevated NEFAs-exposed oocytes (Shi & Sirard, 2021). Enriched pathways of regulated genes and DNA methylation patterns were mainly related to cell activities, metabolism, and inflammation (Shi & Sirard, 2021).…”

mentioning

confidence: 90%

“…In our previous study, altered transcriptome and DNA methylation patterns were found in porcine blastocysts derived from pathologically elevated NEFAs-exposed oocytes (Shi & Sirard, 2021). Enriched pathways of regulated genes and DNA methylation patterns were mainly related to cell activities, metabolism, and inflammation (Shi & Sirard, 2021). However, the role of the close communications between granulosa cells and COCs in vivo was not considered.…”

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confidence: 97%

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Effects of NEFAs during IVM on pig embryos from granulosa cell‐cocultured oocytes

Shi

Sirard

2021

Molecular Reproduction Devel

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Circulating levels of nonesterified fatty acids (NEFAs) are elevated in some females, which can impair oocyte maturation and embryo development, and may alter the phenotype of the progeny. However, the effects of NEFAs on human embryo development are not clear due to ethical limitations. Thus, we used pig as the model to investigate the impacts of NEFAs on oocyte and embryo due to their similar reproductive and metabolic physiologies to humans. In this study, porcine cumulusoocyte complexes were in vitro maturated under a pathologically high concentration of NEFAs (468 μM palmitic acid, 194 μM stearic acid, and 534 μM oleic acid) with the presence of granulosa cell monolayer, in contrast to control without NEFAs. The mature oocytes were fertilized to produce embryos for further analysis of the transcriptome and DNA methylation patterns. The elevated level of NEFAs decreased the blastocyst rate and delayed the blastocyst development. Ingenuity pathway analysis showed that the most affected gene pathways were related mainly to cell activities, metabolism, and inflammation. These findings indicated that oocytes exposed to the exogenous high level of NEFAs during in vitro maturation resulted in altered gene expression and DNA methylation of early embryos, which have detrimental impacts on blastocyst quality.

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Section: Analysis Of Dna Methylation Datamentioning

confidence: 56%

“…Tables 4 and 5 & Sirard, 2021). Therefore, the presence of granulosa cells during maturation seems to lessen the effect of elevated NEFAs.…”

Section: Analysis Of Dna Methylation Datamentioning

confidence: 89%

Section: Analysis Of Dna Methylation Datamentioning

confidence: 99%

mentioning

confidence: 90%

mentioning

confidence: 97%

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Effects of NEFAs during IVM on pig embryos from granulosa cell‐cocultured oocytes

Shi

Sirard

2021

Molecular Reproduction Devel

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Effect of lipotoxicity on mitochondrial function and epigenetic programming during bovine in vitro embryo production

Meulders,

Marei,

Xhonneux

et al. 2023

Sci Rep

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Maternal metabolic disorders may cause lipotoxic effects on the developing oocyte. Understanding the timing at which this might disrupt embryo epigenetic programming and how this is linked with mitochondrial dysfunction is crucial for improving assisted reproductive treatments, but has not been investigated before. Therefore, we used a bovine in vitro model to investigate if pathophysiological palmitic acid (PA) concentrations during in vitro oocyte maturation and in vitro embryo culture alter embryo epigenetic patterns (DNA methylation (5mC) and histone acetylation/methylation (H3K9ac/H3K9me2)) compared to control (CONT) and solvent control (SCONT), at the zygote and morula stage. Secondly, we investigated if these epigenetic alterations are associated with mitochondrial dysfunction and changes in ATP production rate, or altered expression of epigenetic regulatory genes. Compared to SCONT, H3K9ac and H3K9me2 levels were increased in PA-derived zygotes. Also, 5mC and H3K9me2 levels were increased in PA-exposed morulae compared to SCONT. This was associated with complete inhibition of glycolytic ATP production in oocytes, increased mitochondrial membrane potential and complete inhibition of glycolytic ATP production in 4-cell embryos and reduced SOD2 expression in PA-exposed zygotes and morulae. For the first time, epigenetic alterations in metabolically compromised zygotes and morulae have been observed in parallel with mitochondrial dysfunction in the same study.

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Cocultured porcine granulosa cells respond to excess non-esterified fatty acids during in vitro maturation

Shi

Sirard

2021

J Ovarian Res

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Background Non-esterified fatty acids (NEFAs) are one of the main lipid components of follicular fluid at concentrations that depend on circulating levels. Elevated levels of NEFAs impair oocyte quality, development potential, and may subsequently influence the metabolism and reproductive fitness of offspring. Granulosa cells (GCs) are the follicular cells that are closely communicating with the oocyte. However, the responses of GCs exposed to high levels of NEFAs when cocultured with cumulus-oocyte complexes (COCs), and how they attenuate the negative effects of NEFAs on oocytes, are unclear. Results To better understand this protective effect, monolayers of porcine GCs were cocultured with COCs during in vitro maturation (IVM) in the presence of elevated levels of NEFAs. Genomic expression analysis was conducted to explore the responses of the GCs to the elevated levels of NEFAs. After limma algorithm analysis, 1,013 genes were differentially expressed between GCs cultured with and without elevated NEFAs. Among them, 438 genes were upregulated and 575 were downregulated. The differentially expressed genes were enriched in pathways related to metabolism, inflammation, and epithelial-mesenchymal transition. Conclusions The pathways and upstream regulators suggested that the cocultured GCs responded to the elevated NEFAs with (1) inhibition of the transition from granulosa to luteal cell, (2) interactions of metabolism change, anti-inflammation, mitochondrial function, and cell transition, (3) intercommunication with cocultured COCs of anti-inflammatory factors.

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Transcriptome and epigenome analysis of porcine embryos from non-esterified fatty acid-exposed oocytes

Cited by 6 publications

References 55 publications

Effects of NEFAs during IVM on pig embryos from granulosa cell‐cocultured oocytes

Effects of NEFAs during IVM on pig embryos from granulosa cell‐cocultured oocytes

Effect of lipotoxicity on mitochondrial function and epigenetic programming during bovine in vitro embryo production

Cocultured porcine granulosa cells respond to excess non-esterified fatty acids during in vitro maturation

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