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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

Genomic distribution of sperm-derived DMRs induced by prenatal exposure to DEHP in susceptible and resistant F1 mice.(A) Pie chart showing the distribution in % of 2.2 kb sized generated probes among five determined genomic features, exons (light gray), promoters (black), intergenic regions (dark gray), enhancer (red) and introns (white). A total of 1’067’283 starting probes covering 84% of the genome showed an original genomic assignment-based repartition as following; 54% (573'396) intergenic regions, 29% (310’194) introns, 13% (142’843) exons, 0.06% enhancers (684) and 4% (40’166) promoters. (B) Pie charts reflecting the relative distribution expressed in percentages of 2.2 kb sized probes among five determined genomic features that showed statistically significant (p-value below 0.01) either two-fold or more hyper-methylation (the two pie charts located above the horizontal dashed line), or two-fold or more hypo-methylation (the two pie charts located under the horizontal dashed line) when comparing DEHP treated mice to controls. Both backgrounds are analyzed separately, FVB/N on the left of the vertical dashed line, C57BL/6J on the right of the vertical dashed line. The number of probes and the coverage in % of the genome is shown for each of the four tested conditions separated by both dashed lines. The repartitions of probes that did not show DMRs in both strains are not shown; were 55% intergenic, 29% intronic, 12% exonic, 4% promoter and 0.06% enhancer in FVB/N and respectively 53%, 29%, 14%, 4% and 0.06% in C57BL/6J strains. These repartition of probes not affected by DMRs respected the original repartition of probes (A).
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pone.0132136.g002: Genomic distribution of sperm-derived DMRs induced by prenatal exposure to DEHP in susceptible and resistant F1 mice.(A) Pie chart showing the distribution in % of 2.2 kb sized generated probes among five determined genomic features, exons (light gray), promoters (black), intergenic regions (dark gray), enhancer (red) and introns (white). A total of 1’067’283 starting probes covering 84% of the genome showed an original genomic assignment-based repartition as following; 54% (573'396) intergenic regions, 29% (310’194) introns, 13% (142’843) exons, 0.06% enhancers (684) and 4% (40’166) promoters. (B) Pie charts reflecting the relative distribution expressed in percentages of 2.2 kb sized probes among five determined genomic features that showed statistically significant (p-value below 0.01) either two-fold or more hyper-methylation (the two pie charts located above the horizontal dashed line), or two-fold or more hypo-methylation (the two pie charts located under the horizontal dashed line) when comparing DEHP treated mice to controls. Both backgrounds are analyzed separately, FVB/N on the left of the vertical dashed line, C57BL/6J on the right of the vertical dashed line. The number of probes and the coverage in % of the genome is shown for each of the four tested conditions separated by both dashed lines. The repartitions of probes that did not show DMRs in both strains are not shown; were 55% intergenic, 29% intronic, 12% exonic, 4% promoter and 0.06% enhancer in FVB/N and respectively 53%, 29%, 14%, 4% and 0.06% in C57BL/6J strains. These repartition of probes not affected by DMRs respected the original repartition of probes (A).

Mentions: Whole genome analysis of DNA sperm methylation using MBD-Seq indicated that prenatal exposure to DEHP has a higher impact on the fraction of hyper-methylated DMRs in the C57BL/6J strain compared to the FVB/N strain (16% compared with 10%) and less impact on the fraction of hypo-methylated DMRs (3% compared with 5%, respectively) (Fig 2). More specifically, following DEHP-exposure 16% of probes were hyper-methylated and 3% were hypo-methylated in the C57BL/6J strain as compared to 9.7% and 5% respectively in the FVB/N strain. DEHP effects were not randomly distributed in the genome with a secondary finding of approximately three times more exons-assigned probes within hypo-methylated DMRs (34% in FVB/N and 39% in C57Bl/6J compared with 13% at the origin) combined with approximately two times less promoter-assigned probes within hyper-methylated DMRs in both backgrounds (2% in both backgrounds compared with 4% at the origin).


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)

Genomic distribution of sperm-derived DMRs induced by prenatal exposure to DEHP in susceptible and resistant F1 mice.(A) Pie chart showing the distribution in % of 2.2 kb sized generated probes among five determined genomic features, exons (light gray), promoters (black), intergenic regions (dark gray), enhancer (red) and introns (white). A total of 1’067’283 starting probes covering 84% of the genome showed an original genomic assignment-based repartition as following; 54% (573'396) intergenic regions, 29% (310’194) introns, 13% (142’843) exons, 0.06% enhancers (684) and 4% (40’166) promoters. (B) Pie charts reflecting the relative distribution expressed in percentages of 2.2 kb sized probes among five determined genomic features that showed statistically significant (p-value below 0.01) either two-fold or more hyper-methylation (the two pie charts located above the horizontal dashed line), or two-fold or more hypo-methylation (the two pie charts located under the horizontal dashed line) when comparing DEHP treated mice to controls. Both backgrounds are analyzed separately, FVB/N on the left of the vertical dashed line, C57BL/6J on the right of the vertical dashed line. The number of probes and the coverage in % of the genome is shown for each of the four tested conditions separated by both dashed lines. The repartitions of probes that did not show DMRs in both strains are not shown; were 55% intergenic, 29% intronic, 12% exonic, 4% promoter and 0.06% enhancer in FVB/N and respectively 53%, 29%, 14%, 4% and 0.06% in C57BL/6J strains. These repartition of probes not affected by DMRs respected the original repartition of probes (A).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0132136.g002: Genomic distribution of sperm-derived DMRs induced by prenatal exposure to DEHP in susceptible and resistant F1 mice.(A) Pie chart showing the distribution in % of 2.2 kb sized generated probes among five determined genomic features, exons (light gray), promoters (black), intergenic regions (dark gray), enhancer (red) and introns (white). A total of 1’067’283 starting probes covering 84% of the genome showed an original genomic assignment-based repartition as following; 54% (573'396) intergenic regions, 29% (310’194) introns, 13% (142’843) exons, 0.06% enhancers (684) and 4% (40’166) promoters. (B) Pie charts reflecting the relative distribution expressed in percentages of 2.2 kb sized probes among five determined genomic features that showed statistically significant (p-value below 0.01) either two-fold or more hyper-methylation (the two pie charts located above the horizontal dashed line), or two-fold or more hypo-methylation (the two pie charts located under the horizontal dashed line) when comparing DEHP treated mice to controls. Both backgrounds are analyzed separately, FVB/N on the left of the vertical dashed line, C57BL/6J on the right of the vertical dashed line. The number of probes and the coverage in % of the genome is shown for each of the four tested conditions separated by both dashed lines. The repartitions of probes that did not show DMRs in both strains are not shown; were 55% intergenic, 29% intronic, 12% exonic, 4% promoter and 0.06% enhancer in FVB/N and respectively 53%, 29%, 14%, 4% and 0.06% in C57BL/6J strains. These repartition of probes not affected by DMRs respected the original repartition of probes (A).
Mentions: Whole genome analysis of DNA sperm methylation using MBD-Seq indicated that prenatal exposure to DEHP has a higher impact on the fraction of hyper-methylated DMRs in the C57BL/6J strain compared to the FVB/N strain (16% compared with 10%) and less impact on the fraction of hypo-methylated DMRs (3% compared with 5%, respectively) (Fig 2). More specifically, following DEHP-exposure 16% of probes were hyper-methylated and 3% were hypo-methylated in the C57BL/6J strain as compared to 9.7% and 5% respectively in the FVB/N strain. DEHP effects were not randomly distributed in the genome with a secondary finding of approximately three times more exons-assigned probes within hypo-methylated DMRs (34% in FVB/N and 39% in C57Bl/6J compared with 13% at the origin) combined with approximately two times less promoter-assigned probes within hyper-methylated DMRs in both backgrounds (2% in both backgrounds compared with 4% at the origin).

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