Additional file 4: of Expression of epigenetic machinery genes is sensitive to maternal obesity and weight loss in relation to fetal growth in mice

Expression level and adjusted P values of 92 genes in the fetal liver, placental labyrinth, and junctional zone. We assessed the expression level of 60 epigenetic machinery genes and 32 genes implicated in metabolism or development in CTRL, OB, and WL females at E18.5 using TaqMan low-density arrays. Data are represented as mean expression levels ± St.Dev. *—differentially expressed genes. NA—non-amplified. Significant differences (P adj < 0.05) are indicated in red

Additional file 1: of Expression of epigenetic machinery genes is sensitive to maternal obesity and weight loss in relation to fetal growth in mice

Supplementary Figures S1 to S6. Food intake (FI) to body weight (BW) ratio in females during the preconceptional period. (a) P < 0.05 OB vs. CTRL, (b) P < 0.05 WL vs. CTRL, (c) P < 0.05 WL vs. OB. n = 18–20 CTRL, 23 OB, 17–19 WL. Figure S2. Fetal weight as a function of litter size in dams at E18.5. Both sexes were combined as there was no effect of sex on fetal weight. Figure S3. Placental weight as a function of litter size and sex at E18.5. Figure S4. Effect of maternal age on fetal parameters. (A) Relationship between maternal age and fetal weight. (B) Relationship between maternal age and fetal-weight-to-placental-weight ratio index (FPI). For statistical analysis, see text. Figure S5. Distribution of fetal weight in CTRL, OB, and WL dams at E18.5. CTRL dams are represented in black, WL dams in brown, and OB dams in blue. The red line represents the 10th percentile of CTRL population. Figure S6. Relationship between fetal and placental weight in female and male offspring at E18.5. For statistical analysis, see text and “Methods” section. M: males, F: females

Additional file 3: of Expression of epigenetic machinery genes is sensitive to maternal obesity and weight loss in relation to fetal growth in mice

Description of the selection criteria of genes for the custom TLDA design and related bibliographic references

Additional file 2: of Expression of epigenetic machinery genes is sensitive to maternal obesity and weight loss in relation to fetal growth in mice

List of the gene symbol and name, and the design strand and TaqMan® Gene Expression Assay IDentification reference for the 96 genes studied on the Custom TLDA

Schematic diagram summarizing the transcriptional data obtained for the liver, muscle and adipose tissue and the results of epigenetic studies in OP1, OP2 and OR2 mice.

<p>Blue arrows represent potential lipid fluxes. In the hyperphagic OP1 and OP2 mice, lipid ingestion was much greater than in OR2 mice, in which caloric intake was normalized. In OP mice, excess lipids were initially stored in the adipose tissue, leading to adipocyte hypertrophy in OP1 mice and hyperplasia in OP2 mice, as a function of the level of expression of <i>Lep</i> and <i>Peg1</i>. In OR2 mice, lipids were stored in the adipose tissue without adipocyte abnormalities or ectopic storage, as in mice supplied with limited amounts of lipid. In OP1 mice, the excess lipids were stored in the liver, contributing to hepatic hypertrophy. Transcriptomic data indicated that <i>de novo</i> lipogenesis was activated in OP1 mice and that insulin signaling was greatly disturbed. In OP2 mice, genes related to insulin signaling were less affected, whereas genes involved in fatty acid oxidation were globally upregulated. Changes to hepatic metabolism, together with the probable redirection of lipids to muscle thus spared the liver from lipid accumulation. Finally, in OR2 mice, lipid metabolism as a whole was downregulated, whereas thyroid hormone signaling was upregulated. The HFD response was also associated, in OP1 mice, with an upregulation of potassium channels and serotonin receptors, subsequently reversed in both OP2 and OR2 mice. Changes in DNA methylation were observed in the livers of OP1 mice and the muscle of OP2 mice. In the liver, <i>Set7/9</i> expression was decreased by the HFD in mice born to either lean or obese/diabetic mothers (F1LM and F2OM), whether OP or OR. In the livers of OP1 mice, DNA hypomethylation was associated with an upregulation of <i>Suv39h1</i> and <i>Suv39h2</i> expression, whereas, in both OR2 and OP2 mice, normal DNA methylation was associated with a decrease in <i>Jhdm2a</i> expression and an increase in the level of <i>Dnmt2</i> mRNA. In muscle, normal DNA methylation was associated with an upregulation of <i>Suv39h1, Set7/9</i> and <i>Dnmt2</i> in OP1 and OR2 mice, contrasting with the lower level of expression of the <i>Set7/9</i> and <i>Dnmt2</i> genes in the muscle of OP2 mice presenting DNA hypomethylation.</p

Analysis, by RT-qPCR, of the expression of genes encoding histone-modifying enzymes in the liver and muscle of mice fed the HFD, for both lean and obese mothers.

<p>The values shown are the ratios between OP1 or OR1 and CD1 and between OP2 or OR2 and CD2. They are expressed as the mean±SEM, <i>n</i> = 8 per group. ^a<i>p</i><0.05 for comparison with control CD, ^b<i>p</i><0.05 for comparison between OP and OR mice, ^c<i>p</i><0.05 for comparison between the F1LM and F2OM (maternal effect: lean versus obese mother), assessed by Kruskal-Wallis tests and <i>post hoc</i> Dunn’s tests.</p

Analysis, by RT-qPCR, of the expression of genes encoding DNA methyltransferase enzymes in the liver and muscle of mice fed the HFD, born to either lean or obese/diabetic mothers (F1LM and F2OM).

<p>The values shown are the ratios between OP1 or OR1 and CD1 and between OP2 or OR2 and CD2. They are expressed as the mean±SEM, <i>n</i> = 8 per group. ^a<i>p</i><0.05 for comparison with control CD, ^b<i>p</i><0.05 for comparison between OP and OR mice, ^c<i>p</i><0.05 for comparison between the F1LM and F2OM born to lean and obese/diabetic mothers, respectively, assessed by Kruskal-Wallis tests and <i>post hoc</i> Dunn’s tests.</p

Analysis of mRNA levels for key adipogenic genes by RT-qPCR on the adipose tissue of mice fed either the CD or the HFD, born to either lean or obese/diabetic mothers (F1LM and F2OM).

<p>The values shown are the ratios between OP1 or OR1 and CD1 and between OP2 or OR2 and CD2 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066816#pone-0066816-g009" target="_blank">Figures 9</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066816#pone-0066816-g010" target="_blank">10</a>). They are expressed as the mean±SEM, <i>n</i> = 6 per group. ^a<i>p</i><0.05 for comparison with control CD, ^b<i>p</i><0.05 for comparison between OP and OR mice, ^c<i>p</i><0.05 for comparison between F1LM and F2OM, assessed by Kruskal-Wallis tests followed by <i>post hoc</i> Dunn’s tests.</p

Phenotypic and metabolic characteristics of the mice after 24 weeks on the CD and HFD, for the F1LM (CD1, OP1 and OR1) and F2OM (CD2, OP2 and OR2) mice.

<p>The values presented are the means±SEM. For weight and caloric intake, <i>n</i> = 50 to 70, for plasma hormone and metabolite determinations, <i>n</i> = 10 to 15, for FA level determination, <i>n</i> = 7, and for Piximus analysis and organ weights, <i>n</i> = 5.</p>a<p><i>p</i><0.05 for comparison with control,</p>b<p><i>p</i><0.05 for comparison between OP and OR mice,</p>c<p><i>p</i><0.05 for comparison between the F1LM and F2OM mice, assessed by Kruskal-Wallis tests followed by Dunn’s <i>post hoc</i> tests.</p

Resistance to the obesogenic effects of the HFD: Major networks identified by IPA analysis of the genes involved in the response and adaptation to HFD of OR2 mice.

<p>(A) These networks were built from the 142 genes differentially expressed between OR2 and CD2 mice (direct comparison OR2/CD2). Only the first two networks are represented in (B) and (C). The node color indicates the level of expression of the genes: red, upregulated; green, downregulated.</p