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Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation
Published on: June 17, 2015
Impact of mono(2-ethylhexyl) phthalate (MEHP) on the development of mouse embryo in vitro
Rachel Braz Arcanjo1, Marcos Costa Vieira2, Mayandi Sivaguru3
1Department of Animal Sciences, University of Illinois Urbana-Champaign, 1207 West Gregory Drive, Urbana, IL 61801, United States.
Abstract:
Mono(2-ethylhexyl) phthalate (MEHP) is the most studied metabolite of di(2-ethylhexyl) phthalate (DEHP), a phthalate found in cosmetics, flooring, paints, and plastics products, including toys and medical tubing. Humans are frequently exposed to this compound due to its ubiquitous presence in our environment. DEHP and MEHP are known to be endocrine-disrupting chemicals and exposure levels have been associated to decreased reproductive success. However, few studies have focused on the direct effects of MEHP on embryos. The present study investigated effects of MEHP (0.1, 1, 10, 100 and 1000 µM) on mice preimplantation embryonic development, evaluating percentage of blastocyst formation, hatching from zona pellucida, methylation-related genes, cell lineage commitment, micronucleation, and adherens junction marker at different stages of development during in vitro culture for 6 days. We show MEHP negatively impacts embryo competence by reducing blastocyst formation and hatching at 100 and 1000 µM. In addition, 100 µM MEHP increases the expression of Tet3 gene in blastocysts, which is related to a reduction of DNA methylation, an important mechanism regulating gene expression. Exposed embryos that completed the hatching process in groups 0.1, 1 and 10 µM MEHP had similar number of inner cell mass and trophectoderm cells compared to the control, while micronucleation occurrence and E-cadherin expression was not affected in exposed morulae by MEHP at 10 or 100 µM. Our results showed that high concentrations of MEHP can negatively impact embryo development. New studies unveiling the mechanism of toxicity involved and encompassing further developmental stages are warranted for further understanding.
Insights
Mono(2-ethylhexyl) phthalate (MEHP), a metabolite of DEHP, negatively impacts mouse embryo development at high concentrations. Exposure reduced blastocyst formation and hatching, and altered DNA methylation-related gene expression, indicating potential reproductive risks.
Area of Science:
- Reproductive Toxicology
- Developmental Biology
- Environmental Health
Background:
- Di(2-ethylhexyl) phthalate (DEHP) and its metabolite MEHP are ubiquitous endocrine-disrupting chemicals.
- Human exposure to MEHP is common due to its presence in various consumer products.
- MEHP exposure is linked to decreased reproductive success, but its direct effects on early embryonic development are less understood.
Purpose of the Study:
- To investigate the direct effects of MEHP on mouse preimplantation embryonic development in vitro.
- To evaluate MEHP's impact on key developmental milestones and molecular markers.
Main Methods:
- Mouse embryos were cultured in vitro for 6 days with varying concentrations of MEHP (0.1–1000 µM).
- Assessed were blastocyst formation, hatching, DNA methylation-related gene expression (Tet3), cell lineage commitment, micronucleation, and E-cadherin expression.
Main Results:
- MEHP significantly reduced blastocyst formation and hatching rates at concentrations of 100 and 1000 µM.
- A high concentration (100 µM) of MEHP increased Tet3 gene expression in blastocysts, suggesting reduced DNA methylation.
- Lower MEHP concentrations (0.1–10 µM) did not affect cell counts in hatched embryos, nor micronucleation or E-cadherin expression in morulae.
Conclusions:
- High concentrations of MEHP adversely affect critical stages of mouse preimplantation embryonic development.
- MEHP exposure may disrupt embryonic development through mechanisms involving DNA methylation.
- Further research is needed to elucidate the precise toxicity mechanisms and effects on later developmental stages.
