Genes investigated and relevant primer sequences.

<p>Genes investigated and relevant primer sequences.</p

Maternal diet induced obesity increases rates of ROS generation and depletes glutathione in oocytes and zygotes.

<p>Cytosolic ROS production in oocytes and zygotes was measured by rate of oxidation of HEt. The traces represent changes of HEt fluorescence in oocytes (A) and zygotes (B) from lean (n = 15 cells/group) and obese (n = 15 cells/group) mice as a function of time. These data are summarised in histograms (C, D), in which the mean rates of ROS production are shown as the mean rate of HEt fluorescence change per minute. Results are expressed as percentage changes from HEt fluorescence in lean oocytes. Intracellular glutathione staining with MCB in oocytes (E) and zygotes (F) recovered from lean and obese female mice. Relative intensity of MCB fluorescence is expressed as a percentage of the signal from oocytes of lean mice. (G) Representative confocal images of GSH staining in oocytes. *<i>p</i><0.05, ** <i>p</i><0.01. Data are mean ± SEM.</p

Maternal weights and metabolic parameters.

<p>Data expressed as mean ± SEM. All serum measurements were fasting. Values in parentheses indicate n/group.</p

Mitochondrial biogenesis is up-regulated in oocytes from obese mice.

<p>(A) mtDNA copy number in oocytes from lean (n = 2 oocytes/8 females/group) and obese (n = 2 oocytes/8 females/group) mice. Relative abundance of <i>TFAM</i> mRNA (B) and <i>NR1m</i>RNA (C) in oocytes from lean (n = 20 oocytes/8 females/group) and obese (n = 20 oocytes/8 females/group) mice. qPCR was used to determine the absolute mtDNA copy number and the amount of specific transcripts relative to the <i>H2A</i>mRNA.* <i>p</i><0.05. Data are mean ± SEM.</p

Principal maternal outcomes by level of proteinuria adjusted for age and ethnicity with risk ratios in comparison with PE 300 group (proteinuria 300- 499 mg/24 hours).

<p>Data are given as mean (standard deviation) or number (percentage) together with risk ratios and 95% confidence intervals (CI).</p><p>Risk ratio, p in comparison with preeclampsia with proteinuria of 300 - 499 mg/24 hours; *P≤0.05; **P≤0.01; ***P≤0.001 </p

The influence of maternal diet –induced obesity on early embryo development.

<p>Data expressed as mean ± SEM. The number of zygotes was determined in the morning after natural mating. Blastocysts were recovered from the uterus on day 4 after mating. The number of cells per blastocyst and their distribution between the inner cell mass and the trophectoderm were analysed in 3–5 blastocysts per mouse. Values in parentheses indicate a number of mice/group.</p>1<p>In both control and in 2 out of 5 obese mothers a small number of fragmented embryos at various stages of development was recovered from the oviducts.</p

Maternal diet-induced obesity is associated with an oxidised intracellular redox state in oocytes and zygotes.

<p>The redox state of single oocytes and zygotes from lean (n = 15 cells/group) and obese (n = 15 cells/group) mice was estimated through measurements of NAD(P)H and FAD²⁺ autofluorescence intensity. The resting redox state is defined as a ratio of the maximally oxidised (response to 1 mM FCCP) and maximally reduced (response to 1 mM NaCN) signals. The fluorescence signals are normalised between 100 and 0. For NAD(P)H autofluorescence (A): 0 - maximally oxidised state; 100 – maximally reduced state. This scale is reversed for FAD²⁺ fluorescence (B). 0 – maximally reduced state; 100 – maximally oxidised state. * p<0.05 relative lean group. Data are mean ± SEM.</p

AgRP- and α-MSH-immunoreactivity in the PVH of offspring of control and obese dams.

<p>Representative brightfield images (A) and quantitative comparisons (B,C) of AgRP-immunoreactivity in the paraventricular hypothalamic nucleus (PVH) and its subdivisions in male offspring of control and obese dams. Representative confocal images (D) and quantitative comparisons (D,E) of AgRP immunofluorescence in the PVH of female offspring of control and obese dams. Representative brightfield images (F) and quantitative comparisons (G,H) of α-MSH-immunoreactivity in the PVH and its subdivisions in male offspring of control and obese dams. OffCon = offspring of control dams (open bars); OffOb = offspring of obese dams (closed bars); mp = medial parvocellular; lm = lateral magnocellular; dp = dorsal parvocellular. * = p<0.05 and ** = p<0.01 <i>versus</i> control. Scale bars = 200 µm.</p

Behavioural responses to leptin in juvenile and adult offspring of control and obese dams.

<p>Food intake for males (A) and females (B) and change in body weight for males (C) and females (D) recorded over 24 hours following administration of leptin (10 mg/kg, i.p.) in 30 day-old offspring of control or obese dams. Food intake for males (E) and females (F) and change in body weight for males (G) and females (H) in 90 day-old offspring of control or obese dams. OffCon = offspring of control dams; OffOb = offspring of obese dams; * p<0.05 and ** p<0.01 ***p<0.001, <i>versus</i> offspring of control dams (n = 6).</p

Signalling responses to leptin in juvenile and adult offspring of control and obese dams.

<p>Representative images (A) and quantitative analysis (B and C) of leptin-induced pSTAT3-immunoreactive (ir) cells (per mm²) in the arcuate nucleus (ARC) and ventromedial hypothalamic nucleus (VMH). Leptin (10 mg/kg, i.p.) was administered 45 min prior to administration of anaesthetic and perfusion fixation of the brain. OffCon = offspring of control dams; OffOb = offspring of obese dams; dl = dorsolateral; vl = ventrolateral. * p<0.05 <i>versus</i> offspring of control dams (n = 4–8).</p