Limits...
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

DEHP-induced DMRs in sperm promoters.Volcano plots illustrating promoter methylations in function of the prenatal exposure to DEHP in sperm in both backgrounds. The data were obtained from 5 control mice compared with 5 DEHP treated mice for each background, FVB/N shown left and C57BL/6J shown right. X axis represent the log2 fold change between controls and DEHP. Y axis represents the minus log10 of the p-value. Horizontal dashed lines represent the genome-wide statistical significance level settled at 2.2*10−6 (0.05 alpha divided by 22’480 tested probes). Vertical lines represent biological significance thresholds for DMRs settled above 1 and under -1 on the x-axes, corresponding respectively to two-fold methylation increase in controls compared with DEHP-treated and two-fold methylation decrease in controls compared with DEHP-treated. Blue points represent promoter showing statistically and biologically significant increased methylations in DEHP, whereas red points represent promoter showing statistically and biologically significant increased methylation in controls. The total of blue and red points are indicates on each graph using blue and red numbers. The number of promoter showing a statistically not significant more than two-fold increase methylation level in DEHP condition is indicates in black on the bottom left of each graph, whereas the number of promoter showing a statistically not significant more of than two-fold decreased methylation in DEHP compared to control is indicates in black on the bottom right of each graph. (A) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering 22’480 different promoters, each assigned to a different murine gene. The most significant point corresponds to the promoter of Sfi1 and is indicated. (B) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 238 vomeronasal receptors whose gene symbols contains “Vmn”. (C) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 1074 olfactory receptors whose gene symbols contains “Olfr”. (D) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 996 micro-RNAs whose gene symbols contains “Mir”.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4526524&req=5

pone.0132136.g003: DEHP-induced DMRs in sperm promoters.Volcano plots illustrating promoter methylations in function of the prenatal exposure to DEHP in sperm in both backgrounds. The data were obtained from 5 control mice compared with 5 DEHP treated mice for each background, FVB/N shown left and C57BL/6J shown right. X axis represent the log2 fold change between controls and DEHP. Y axis represents the minus log10 of the p-value. Horizontal dashed lines represent the genome-wide statistical significance level settled at 2.2*10−6 (0.05 alpha divided by 22’480 tested probes). Vertical lines represent biological significance thresholds for DMRs settled above 1 and under -1 on the x-axes, corresponding respectively to two-fold methylation increase in controls compared with DEHP-treated and two-fold methylation decrease in controls compared with DEHP-treated. Blue points represent promoter showing statistically and biologically significant increased methylations in DEHP, whereas red points represent promoter showing statistically and biologically significant increased methylation in controls. The total of blue and red points are indicates on each graph using blue and red numbers. The number of promoter showing a statistically not significant more than two-fold increase methylation level in DEHP condition is indicates in black on the bottom left of each graph, whereas the number of promoter showing a statistically not significant more of than two-fold decreased methylation in DEHP compared to control is indicates in black on the bottom right of each graph. (A) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering 22’480 different promoters, each assigned to a different murine gene. The most significant point corresponds to the promoter of Sfi1 and is indicated. (B) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 238 vomeronasal receptors whose gene symbols contains “Vmn”. (C) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 1074 olfactory receptors whose gene symbols contains “Olfr”. (D) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 996 micro-RNAs whose gene symbols contains “Mir”.

Mentions: The number of promoters significantly methylated in DEHP-exposed mice shown as blue points was higher in the C57BL/6J strain (n = 231) compared to FVB/N (n = 60), whereas more de-methylated promoters appearing as red dots were presents in FVB/N (n = 418) compared to C57BL/6J strain (n = 242), (Fig 3A). The most significant DMR recorded was Sfi1 promoter hyper-methylated in controls in both backgrounds (Fig 3A, highest red points in both volcano plots). Three main groups of genes were identified by high occurrences of gene names beginning with “vmn”, “olfr” and “mir” in DEHP-induced significant DMRs. The two first consist of genes encoding vomeronasal (Vmn) and olfactory (Olfr) receptors. Massive and global trends to be hyper-methylated in DEHP conditions were observed for 55% ((129+1)/238) of all the vmn promoters in FVB/N and respectively for 74% ((161+14) /238) in C57BL/6J, with similar trends for 54% ((576+7)/1074 of all olfr genes in FVB/N and for 69% ((698+38) /1074) in C57BL/6J, whereas bucking events comparatively remains rare (n = 10, 1, 19 and 3) (Fig 3BC). Higher numbers of those significant targets are observed in C57BL/6J strain compared to FVB/N, respectively 14 and 38 compared with 1 and 7 (Fig 3BC). DEHP conditions had more pronounced statistically significant and opposite impacts on micro-RNAs’ promoters, this class represents almost a third of all the significant hypo-methylated genes in both strains (Fig 3AD) with a higher number of significant hypo-methylated promoter in FVB/N compared to C57BL/6J, 121 versus 67 (Fig 3D).


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)

DEHP-induced DMRs in sperm promoters.Volcano plots illustrating promoter methylations in function of the prenatal exposure to DEHP in sperm in both backgrounds. The data were obtained from 5 control mice compared with 5 DEHP treated mice for each background, FVB/N shown left and C57BL/6J shown right. X axis represent the log2 fold change between controls and DEHP. Y axis represents the minus log10 of the p-value. Horizontal dashed lines represent the genome-wide statistical significance level settled at 2.2*10−6 (0.05 alpha divided by 22’480 tested probes). Vertical lines represent biological significance thresholds for DMRs settled above 1 and under -1 on the x-axes, corresponding respectively to two-fold methylation increase in controls compared with DEHP-treated and two-fold methylation decrease in controls compared with DEHP-treated. Blue points represent promoter showing statistically and biologically significant increased methylations in DEHP, whereas red points represent promoter showing statistically and biologically significant increased methylation in controls. The total of blue and red points are indicates on each graph using blue and red numbers. The number of promoter showing a statistically not significant more than two-fold increase methylation level in DEHP condition is indicates in black on the bottom left of each graph, whereas the number of promoter showing a statistically not significant more of than two-fold decreased methylation in DEHP compared to control is indicates in black on the bottom right of each graph. (A) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering 22’480 different promoters, each assigned to a different murine gene. The most significant point corresponds to the promoter of Sfi1 and is indicated. (B) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 238 vomeronasal receptors whose gene symbols contains “Vmn”. (C) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 1074 olfactory receptors whose gene symbols contains “Olfr”. (D) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 996 micro-RNAs whose gene symbols contains “Mir”.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0132136.g003: DEHP-induced DMRs in sperm promoters.Volcano plots illustrating promoter methylations in function of the prenatal exposure to DEHP in sperm in both backgrounds. The data were obtained from 5 control mice compared with 5 DEHP treated mice for each background, FVB/N shown left and C57BL/6J shown right. X axis represent the log2 fold change between controls and DEHP. Y axis represents the minus log10 of the p-value. Horizontal dashed lines represent the genome-wide statistical significance level settled at 2.2*10−6 (0.05 alpha divided by 22’480 tested probes). Vertical lines represent biological significance thresholds for DMRs settled above 1 and under -1 on the x-axes, corresponding respectively to two-fold methylation increase in controls compared with DEHP-treated and two-fold methylation decrease in controls compared with DEHP-treated. Blue points represent promoter showing statistically and biologically significant increased methylations in DEHP, whereas red points represent promoter showing statistically and biologically significant increased methylation in controls. The total of blue and red points are indicates on each graph using blue and red numbers. The number of promoter showing a statistically not significant more than two-fold increase methylation level in DEHP condition is indicates in black on the bottom left of each graph, whereas the number of promoter showing a statistically not significant more of than two-fold decreased methylation in DEHP compared to control is indicates in black on the bottom right of each graph. (A) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering 22’480 different promoters, each assigned to a different murine gene. The most significant point corresponds to the promoter of Sfi1 and is indicated. (B) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 238 vomeronasal receptors whose gene symbols contains “Vmn”. (C) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 1074 olfactory receptors whose gene symbols contains “Olfr”. (D) Volcano plots of methylation results based on normalized reads numbers that were recorded in 2.2kb sized probes covering a total of 996 micro-RNAs whose gene symbols contains “Mir”.
Mentions: The number of promoters significantly methylated in DEHP-exposed mice shown as blue points was higher in the C57BL/6J strain (n = 231) compared to FVB/N (n = 60), whereas more de-methylated promoters appearing as red dots were presents in FVB/N (n = 418) compared to C57BL/6J strain (n = 242), (Fig 3A). The most significant DMR recorded was Sfi1 promoter hyper-methylated in controls in both backgrounds (Fig 3A, highest red points in both volcano plots). Three main groups of genes were identified by high occurrences of gene names beginning with “vmn”, “olfr” and “mir” in DEHP-induced significant DMRs. The two first consist of genes encoding vomeronasal (Vmn) and olfactory (Olfr) receptors. Massive and global trends to be hyper-methylated in DEHP conditions were observed for 55% ((129+1)/238) of all the vmn promoters in FVB/N and respectively for 74% ((161+14) /238) in C57BL/6J, with similar trends for 54% ((576+7)/1074 of all olfr genes in FVB/N and for 69% ((698+38) /1074) in C57BL/6J, whereas bucking events comparatively remains rare (n = 10, 1, 19 and 3) (Fig 3BC). Higher numbers of those significant targets are observed in C57BL/6J strain compared to FVB/N, respectively 14 and 38 compared with 1 and 7 (Fig 3BC). DEHP conditions had more pronounced statistically significant and opposite impacts on micro-RNAs’ promoters, this class represents almost a third of all the significant hypo-methylated genes in both strains (Fig 3AD) with a higher number of significant hypo-methylated promoter in FVB/N compared to C57BL/6J, 121 versus 67 (Fig 3D).

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