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Prenatal Exposure to DEHP Affects Spermatogenesis and Sperm DNA Methylation in a Strain-Dependent Manner.

Prados J, Stenz L, Somm E, Stouder C, Dayer A, Paoloni-Giacobino A - PLoS ONE (2015)

Bottom Line: Di-(2-ethylhexyl)phtalate (DEHP) is a plasticizer with endocrine disrupting properties found ubiquitously in the environment and altering reproduction in rodents.The number of differentially methylated regions (DMRs) by DEHP-exposure across the entire genome showed increased hyper- and decreased hypo-methylation in C57BL/6J compared to FVB/N.In contrast, a large set of micro-RNAs were hypo-methylated, with a trend more pronounced in the FVB/N strain.

View Article: PubMed Central - PubMed

Affiliation: Department of Mental Health and Psychiatry, Division of Psychiatric Specialties, University Hospitals of Geneva, Geneva, Switzerland; Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland.

ABSTRACT
Di-(2-ethylhexyl)phtalate (DEHP) is a plasticizer with endocrine disrupting properties found ubiquitously in the environment and altering reproduction in rodents. Here we investigated the impact of prenatal exposure to DEHP on spermatogenesis and DNA sperm methylation in two distinct, selected, and sequenced mice strains. FVB/N and C57BL/6J mice were orally exposed to 300 mg/kg/day of DEHP from gestation day 9 to 19. Prenatal DEHP exposure significantly decreased spermatogenesis in C57BL/6J (fold-change = 0.6, p-value = 8.7*10-4), but not in FVB/N (fold-change = 1, p-value = 0.9). The number of differentially methylated regions (DMRs) by DEHP-exposure across the entire genome showed increased hyper- and decreased hypo-methylation in C57BL/6J compared to FVB/N. At the promoter level, three important subsets of genes were massively affected. Promoters of vomeronasal and olfactory receptors coding genes globally followed the same trend, more pronounced in the C57BL/6J strain, of being hyper-methylated in DEHP related conditions. In contrast, a large set of micro-RNAs were hypo-methylated, with a trend more pronounced in the FVB/N strain. We additionally analyze both the presence of functional genetic variations within genes that were associated with the detected DMRs and that could be involved in spermatogenesis, and DMRs related with the DEHP exposure that affected both strains in an opposite manner. The major finding in this study indicates that prenatal exposure to DEHP can decrease spermatogenesis in a strain-dependent manner and affects sperm DNA methylation in promoters of large sets of genes putatively involved in both sperm chemotaxis and post-transcriptional regulatory mechanisms.

No MeSH data available.


Related in: MedlinePlus

Impact of promoter methylation on gene expressions in pre-selected candidates.Graphical representation of the obtained MBDseq and RT-qPCR results on a subset of pre-selected genes and in respect with the mouse strain background, C57BL/6J on the left part of the figure and FVB on the right side of the figure. The x-axis showed results obtained between the control condition (CTL) always on the left and the DEHP 300 condition (DEHP) always on the right of each graphic as mentioned. Levels of mRNA appear as gray dots and are expressed as arbitrary units on the left-side Y-axis labelled in gray and named mRNA levels (A.U.). A.U.: Arbitrary units relative to the control condition. The MBD-seq promoter methylation levels appear as black squares and are expressed in average of reads for each condition labelled on the right-side Y-axis in each graphic. Errors bars represent standard error of measurements for mRNA levels and standards deviation for MBD-seq derived reads averages. Inverted slopes between MBDseq and mRNA data are interpreted as results consistent with a negative impact of promoter methylation on gene expression. Stars in gray represent significant differences in term of mRNA levels obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05. Stars in black represent statistically significant differences in term of average reads numbers obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05.
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pone.0132136.g006: Impact of promoter methylation on gene expressions in pre-selected candidates.Graphical representation of the obtained MBDseq and RT-qPCR results on a subset of pre-selected genes and in respect with the mouse strain background, C57BL/6J on the left part of the figure and FVB on the right side of the figure. The x-axis showed results obtained between the control condition (CTL) always on the left and the DEHP 300 condition (DEHP) always on the right of each graphic as mentioned. Levels of mRNA appear as gray dots and are expressed as arbitrary units on the left-side Y-axis labelled in gray and named mRNA levels (A.U.). A.U.: Arbitrary units relative to the control condition. The MBD-seq promoter methylation levels appear as black squares and are expressed in average of reads for each condition labelled on the right-side Y-axis in each graphic. Errors bars represent standard error of measurements for mRNA levels and standards deviation for MBD-seq derived reads averages. Inverted slopes between MBDseq and mRNA data are interpreted as results consistent with a negative impact of promoter methylation on gene expression. Stars in gray represent significant differences in term of mRNA levels obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05. Stars in black represent statistically significant differences in term of average reads numbers obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05.

Mentions: Expression and methylation in the six pre-selected targets mentioned in the method section are presented on (Fig 6). Results showed inverted slopes between MBDseq and mRNA data suggesting a negative impact of promoter methylation on gene expression.


Prenatal Exposure to DEHP Affects Spermatogenesis and Sperm DNA Methylation in a Strain-Dependent Manner.

Prados J, Stenz L, Somm E, Stouder C, Dayer A, Paoloni-Giacobino A - PLoS ONE (2015)

Impact of promoter methylation on gene expressions in pre-selected candidates.Graphical representation of the obtained MBDseq and RT-qPCR results on a subset of pre-selected genes and in respect with the mouse strain background, C57BL/6J on the left part of the figure and FVB on the right side of the figure. The x-axis showed results obtained between the control condition (CTL) always on the left and the DEHP 300 condition (DEHP) always on the right of each graphic as mentioned. Levels of mRNA appear as gray dots and are expressed as arbitrary units on the left-side Y-axis labelled in gray and named mRNA levels (A.U.). A.U.: Arbitrary units relative to the control condition. The MBD-seq promoter methylation levels appear as black squares and are expressed in average of reads for each condition labelled on the right-side Y-axis in each graphic. Errors bars represent standard error of measurements for mRNA levels and standards deviation for MBD-seq derived reads averages. Inverted slopes between MBDseq and mRNA data are interpreted as results consistent with a negative impact of promoter methylation on gene expression. Stars in gray represent significant differences in term of mRNA levels obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05. Stars in black represent statistically significant differences in term of average reads numbers obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4526524&req=5

pone.0132136.g006: Impact of promoter methylation on gene expressions in pre-selected candidates.Graphical representation of the obtained MBDseq and RT-qPCR results on a subset of pre-selected genes and in respect with the mouse strain background, C57BL/6J on the left part of the figure and FVB on the right side of the figure. The x-axis showed results obtained between the control condition (CTL) always on the left and the DEHP 300 condition (DEHP) always on the right of each graphic as mentioned. Levels of mRNA appear as gray dots and are expressed as arbitrary units on the left-side Y-axis labelled in gray and named mRNA levels (A.U.). A.U.: Arbitrary units relative to the control condition. The MBD-seq promoter methylation levels appear as black squares and are expressed in average of reads for each condition labelled on the right-side Y-axis in each graphic. Errors bars represent standard error of measurements for mRNA levels and standards deviation for MBD-seq derived reads averages. Inverted slopes between MBDseq and mRNA data are interpreted as results consistent with a negative impact of promoter methylation on gene expression. Stars in gray represent significant differences in term of mRNA levels obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05. Stars in black represent statistically significant differences in term of average reads numbers obtained between controls and DEHP 300 conditions according to two-tails T-test resulting in p-value lower than 0.05.
Mentions: Expression and methylation in the six pre-selected targets mentioned in the method section are presented on (Fig 6). Results showed inverted slopes between MBDseq and mRNA data suggesting a negative impact of promoter methylation on gene expression.

Bottom Line: Di-(2-ethylhexyl)phtalate (DEHP) is a plasticizer with endocrine disrupting properties found ubiquitously in the environment and altering reproduction in rodents.The number of differentially methylated regions (DMRs) by DEHP-exposure across the entire genome showed increased hyper- and decreased hypo-methylation in C57BL/6J compared to FVB/N.In contrast, a large set of micro-RNAs were hypo-methylated, with a trend more pronounced in the FVB/N strain.

View Article: PubMed Central - PubMed

Affiliation: Department of Mental Health and Psychiatry, Division of Psychiatric Specialties, University Hospitals of Geneva, Geneva, Switzerland; Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland.

ABSTRACT
Di-(2-ethylhexyl)phtalate (DEHP) is a plasticizer with endocrine disrupting properties found ubiquitously in the environment and altering reproduction in rodents. Here we investigated the impact of prenatal exposure to DEHP on spermatogenesis and DNA sperm methylation in two distinct, selected, and sequenced mice strains. FVB/N and C57BL/6J mice were orally exposed to 300 mg/kg/day of DEHP from gestation day 9 to 19. Prenatal DEHP exposure significantly decreased spermatogenesis in C57BL/6J (fold-change = 0.6, p-value = 8.7*10-4), but not in FVB/N (fold-change = 1, p-value = 0.9). The number of differentially methylated regions (DMRs) by DEHP-exposure across the entire genome showed increased hyper- and decreased hypo-methylation in C57BL/6J compared to FVB/N. At the promoter level, three important subsets of genes were massively affected. Promoters of vomeronasal and olfactory receptors coding genes globally followed the same trend, more pronounced in the C57BL/6J strain, of being hyper-methylated in DEHP related conditions. In contrast, a large set of micro-RNAs were hypo-methylated, with a trend more pronounced in the FVB/N strain. We additionally analyze both the presence of functional genetic variations within genes that were associated with the detected DMRs and that could be involved in spermatogenesis, and DMRs related with the DEHP exposure that affected both strains in an opposite manner. The major finding in this study indicates that prenatal exposure to DEHP can decrease spermatogenesis in a strain-dependent manner and affects sperm DNA methylation in promoters of large sets of genes putatively involved in both sperm chemotaxis and post-transcriptional regulatory mechanisms.

No MeSH data available.


Related in: MedlinePlus