Antioxidant supplementation may effect DNA methylation patterns, apoptosis, and ROS levels in developing mouse embryos

Fatma Uysal ¹, Gozde Sukur ^2,3, Nazlican Bozdemir ¹

⁰Department of Histology and Embryology, Ankara Medipol University School of Medicine, Altindag, 06050, Ankara, Turkey.

Histochemistry and Cell Biology +

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April 16, 2024

View abstract on PubMed

Summary

Antioxidants in embryo culture media impact DNA methylation and cell differentiation. Different antioxidants like NAC, ALC, and MitoQ altered DNA methyltransferases and global methylation, affecting inner cell mass and trophoblast development.

Area Of Science

Reproductive biology
Developmental biology
Epigenetics

Background

Embryo culture media composition is critical for early development.
Antioxidants are explored for their potential to mitigate oxidative stress during in vitro culture.
The impact of specific antioxidants on epigenetic modifications and cell fate decisions in embryos remains incompletely understood.

Purpose Of The Study

To investigate the effects of various antioxidants on DNA methyltransferases (Dnmts), global DNA methylation, inner cell mass/trophoblast differentiation, reactive oxygen species (ROS) levels, and apoptosis in mouse embryos.
To compare the impact of individual antioxidants (MitoQ, N-acetyl-L-cysteine (NAC), acetyl-L-carnitine (ALC), α-lipoic acid (ALA)) and a mixture (NAC + ALC + ALA) on early embryonic development.

Main Methods

Mouse zygotes were cultured in vitro with different antioxidants until the blastocyst stage.
Protein expression of Dnmt1, 3a, 3b, and 3l was analyzed using immunofluorescence and western blot.
Global DNA methylation, ROS levels, and apoptosis were assessed via immunofluorescence.
Nanog and Cdx2 expression were evaluated to assess inner cell mass and trophoblast differentiation, respectively.

Main Results

NAC, ALC, and MitoQ significantly increased Dnmt levels and global DNA methylation.
ALA significantly induced Dnmt expression but did not alter global methylation.
NAC and the antioxidant mixture (AO) increased Nanog levels, while ALA and MitoQ increased Cdx2 levels.
ALA and MitoQ reduced intracellular ROS, whereas ALC increased it.

Conclusions

Antioxidants exert differential effects on DNA methylation and cell differentiation pathways in mouse embryos.
The choice of antioxidant in embryo culture media can influence key developmental markers.
Further research is needed to understand the long-term consequences of these antioxidant-induced epigenetic and developmental alterations on offspring health.

Keywords:

Antioxidants Blastocyst DNA methylation Embryo culture Oxidative stress