Effect of long-term vitamin D supplementation on colonic gene expression in IL-10 Knockout mice at young adulthood
ABSTRACT: Low vitamin D status has been implicated in the progression of inflammatory bowel disease (IBD). This study used interleukin (IL)-10 knockout (KO) mice, that develop an intestinal inflammation when housed in a non-sterile environment, to determine if supplementation with vitamin D throughout life impacts colonic gene expression. Results provide important information on the intestinal response to vitamin D in inflamed mice. Female IL-10 knock out mice were randomized to 25 (Low, L) or 5000 (High, H) IU vitamin D/kg of diet throughout pregnancy and lactation. At weaning, offspring received the same or opposite diet as their mother until age 3 months. This resulted in four vitamin D interventions: HH, HL, LH, or LL where the first letter represents the diet consumed by dams during pregnancy and lactation and the second letter represents the diet consumed by offspring from weaning through to 3 months of age. Global gene expression was analyzed in the proximal colon of 3 months old mice (n=6 per group, for a total of 24 samples; samples came from different litters and moms IDs are given in the samples table below). Samples were stored at -80C.
Project description:Low vitamin D status has been implicated in the progression of inflammatory bowel disease (IBD). This study used interleukin (IL)-10 knockout (KO) mice, that develop an intestinal inflammation when housed in a non-sterile environment, to determine if supplementation with vitamin D throughout life impacts colonic gene expression. Results provide important information on the intestinal response to vitamin D in inflamed mice. Overall design: Female IL-10 knock out mice were randomized to 25 (Low, L) or 5000 (High, H) IU vitamin D/kg of diet throughout pregnancy and lactation. At weaning, offspring received the same or opposite diet as their mother until age 3 months. This resulted in four vitamin D interventions: HH, HL, LH, or LL where the first letter represents the diet consumed by dams during pregnancy and lactation and the second letter represents the diet consumed by offspring from weaning through to 3 months of age. Global gene expression was analyzed in the proximal colon of 3 months old mice (n=6 per group, for a total of 24 samples; samples came from different litters and moms IDs are given in the samples table below). Samples were stored at -80C.
Project description:Initiation of a vitamin A deficient diet in mid-gestation, maintained in the post-weaning diet is sufficient to cause liver and serum retinoid depletion. Wild type C57Bl/6J timed mated pregnant dams were administered either a defined vitamin A sufficient low fat (12 percent kcal from fat) diet or matched vitamin A deficient diet from embryonic day 10.5. Vitamin A sufficient offspring were weaned onto either a high fat diet (60 percent kcal from fat) or maintained on the low fat 12 percent kcal from fat diet for 11 weeks (14 weeks of age). Gestational vitamin A deficient offspring were maintained on the same vitamin A deficient diet until 14 weeks of age. The impact of the maternal diet on a post-weaning high fat diet was compared to a standard maternal breeder diet followed by the post-weaning high fat diet. Overall design: Three comparison experiment. Liver expression of WT C57Bl/6J mice at 14 weeks of age from dams fed a particular maternal diet (X) and weaned onto a post-weaning diet (Y) such that diets are presented X-Y. Breeder diet - High fat diet compared to Low fat 12 percent kcal vitamin A sufficient - High fat diet. Low fat 12 percent kcal vitamin A sufficient - High fat diet compared to Low fat 12 percent kcal vitamin A sufficient - Low fat 12 percent kcal vitamin A sufficient. Low fat 12 percent kcal vitamin A sufficient - Low fat 12 percent kcal vitamin A sufficient compared to Low fat 12 percent kcal vitamin A deficient - Low fat 12 percent kcal vitamin A deficient.
Project description:Background: Epidemiological studies suggest an association between maternal obesity and adverse neurodevelopmental outcomes in offspring. Objective: To compare the global proteomic portrait in the cerebral cortex between mice born to mothers on a high-fat or control diet who themselves were fed a high-fat or control diet. Methods: Male mice born to dams fed a control (C) or high fat (H) diet four weeks before conception and during gestation and lactation were assigned to either C or H diet at weaning. Mice (n=24) were sacrificed at 19-weeks and their cerebral cortices were pooled into 8 samples and analysed using an iTRAQ based 2D LC-MS methodology. Results: A total of 6,695 proteins were identified and fully quantified (q<0.01). Approximately 10% of these proteins demonstrated a minimum of one Standard Deviation of regulation across all biological replicates in at least one of the experimental groups (CH, HC, HH) relative to the control (CC). Principal component analysis and hierarchical clustering analysis showed that mice clustered based on the diet of the mother and not their current diet. In silico bioinformatics analysis revealed that maternal high-fat diet was significantly associated with response to hypoxia/oxidative stress and apoptosis in the cerebral cortex of the adult offspring. Conclusion: Maternal high-fat diet was associated with distinct endophenotypic changes of the adult mouse cerebral cortex independent of the diet of the offspring. The identified modulated proteins could represent novel therapeutic targets for the prevention of neuropathological features resulting from maternal obesity.
Project description:We studied maternal low protein diet (LPD) during pregnancy/lactation in mice. This affected behavioral responses to acute stress in the offspring. We therefore investigated the transcriptome-wide changes following 15 min forced swim, to identify possible immediate early genes affected by perinatal LPD.
Project description:Maternal obesity can program metabolic syndrome in offspring but the mechanisms are not well characterized. Moreover, the consequences of maternal overnutrition in the absence of frank obesity remain poorly understood. This study aimed to determine the effects of maternal consumption of a high fat-sucrose diet on the skeletal muscle metabolic and transcriptional profiles of adult offspring. Female Sprague Dawley rats were fed either a diet rich in saturated fat and sucrose (HFD, 23.5% fat, 20% sucrose wt/wt) or a standard chow diet (NFD, 7% fat, 10% sucrose w/w) for the 3 weeks prior to mating and throughout pregnancy and lactation. Although maternal weights were not different between groups at conception or weaning, HFD dams were ~22% heavier than chow fed dams from mid-pregnancy until 4 days post-partum. Adult male offspring of HFD dams were not heavier than controls but demonstrated features of insulin resistance including elevated plasma insulin concentration (+40%, P<0.05). Next Generation mRNA Sequencing was used to identify differentially expressed genes in the soleus muscle of offspring, and Gene Set Enrichment Analysis (GSEA) to detect coordinated changes that are characteristic of a biological function. GSEA identified 15 pathways enriched for up-regulated genes, including cytokine signaling (P<0.005), starch and sucrose metabolism (P<0.017), and inflammatory response (P<0.024). A further 8 pathways were significantly enriched for down-regulated genes including oxidative phosphorylation (P<0.004) and electron transport (P<0.022). Western blots confirmed a ~60% reduction in the phosphorylation of the insulin signaling protein Akt (P<0.05) and ~70% reduction in mitochondrial complexes II (P<0.05) and V expression (P<0.05). On a normal diet, offspring of HFD dams developed an insulin resistant phenotype, with transcriptional evidence of muscle cytokine activation, inflammation and mitochondrial dysfunction. These data indicate that maternal overnutrition, even in the absence of pre-pregnancy obesity can promote metabolic dysregulation and predispose offspring to type 2 diabetes. Messenger RNA profile of skeletal muscle of male offspring from female Sprague Dawley rats fed either a diet rich in saturated fat and sucrose (HFD, 23.5% fat, 20% sucrose wt/wt) or a standard chow diet (NFD, 7% fat, 10% sucrose w/w) for the 3 weeks prior to mating and throughout pregnancy and lactation. There were 5 HFD samples compared to 6 NFD control samples.
Project description:Maternal high-fat consumption has negative effects on the offspring’s obesity/diabetes susceptibility and we hypothesize that epigenetic modifications in the skeletal muscle are partly responsible for this phenotype. To detect genes affected by maternal nutrition, offspring of low-fat (LF) and high-fat (HF) diet fed dams (C57BL/6 mice) received LF diet upon weaning and were sacrificed at an age of 6 or 25 weeks. M. quadriceps gene expression was investigated by microarray analysis revealing upregulation of the nuclear receptor Nr4a1 by maternal HF feeding. This was accompanied by promoter hypomethylation of CpG‑1408 which correlated with higher Nr4a1 gene expression at both ages. Offspring voluntary exercise training normalized Nr4a1 methylation/expression and ameliorated the negative effects of maternal HF feeding on insulin sensitivity. Overall, Nr4a1 expression correlated with higher insulin levels during oral glucose tolerance test and could, therefore, be involved in the programming offspring’s diabetes susceptibility. Overall design: Starting three days prior to mating, female C57BL/6J mice (8 weeks old) received either a semisynthetic low-fat (LF; 10% energy from fat, 23% energy from protein, 67% energy from carbohydrates) or HF diet (40% energy from fat, 23% energy from protein, 37% energy from carbohydrates) diet during pregnancy and lactation (mLFD and mHFD, resp.). For mating, two females were introduced to one male for two weeks and subsequently individually caged. The litter size was adjusted to 5–7 pups, with no more than four siblings finally present in one group. Mice were kept in a light- and temperature-controlled facility at 22°C with a 12 h light-12 h dark cycle. After weaning, mLF and mHF male offspring were single caged and received LF until the end of the study at 6 weeks of age. All mice were sacrificed after 2 h of fasting using isoflurane and cervical dislocation.
Project description:Wild type C57Bl/6J, Cyp1b1-null, and a substrain of Cyp1b1-null that are resistant to diet-induced obesity (Resistant Cyp1b1-null) timed mated pregnant dams were administered either a defined vitamin A sufficient diet or matched vitamin A deficient diet from embryonic day 4.5. Offspring liver gene expression was examined at birth (post-natal day 0) and at weaning (post-natal day 21). Overall design: Nine comparison experiment. C57Bl/6J WT vitamin A sufficient birth (PN0) control with WT vitamin A deficient PN0 and Cyp1b1-null vitamin A sufficient PN0. Cyp1b1-null vitamin A sufficient PN0 with Cyp1b1-null vitamin A deficient PN0. WT vitamin A sufficient weaning (PN21) control with WT vitamin A deficient PN21 and Cyp1b1-null vitamin A sufficient PN21 and R-Cyp1b1-null vitamin A sufficient PN21. Cyp1b1-null vitamin A sufficient PN21 with Cyp1b1-null vitamin A deficent PN21.R-Cyp1b1-null vitamin A sufficient PN21 with R-Cyp1b1-null vitamin A deficient PN21. WT vitamin A sufficient PN0 with WT vitamin A sufficient PN21.
Project description:Zinc deficiency during pregnancy and postnatal life can adversely affect the health and predispose to an increased risk of developing human diseases at adulthood. The present study was designed to evaluate whether dietary zinc deficiency or supplementation during the pregnancy, lactation and juvenile stages interferes with the development of mammary tumors induced by 7,12-dimethylbenzanthracene (DMBA) in female Sprague-Dawley (SD) rats. Pregnant females SD were allocated into three groups: dams received diets containing adequate (35 mg/Kg chow), deficient (3 mg/Kg chow) or supplemented (180 mg/Kg chow) zinc levels during gestational day 10 (GD 10) until the litters weaning. Female offspring were allocated into three groups and received the same diets as their dams until postnatal day (PND) 51. At PND 51, females SD (n=16 each group) received a single dose of DMBA (50 mg/kg, ig) for initiation of mammary carcinogenesis, adequate zinc diets and were euthanized at PND 180. Tumors samples were collected and processed for histological evaluation and microarray analysis. Dietary zinc deficiency induced a significant reduction in female offspring body weight evolution at parturition, weaning and pubertal phases. At late in life, the early in life dietary zinc deficiency or supplementation did not alter the latency, incidence, multiplicity and volume or histological types of mammary tumors in relation to the adequate zinc group, and changed the expression of a small amount of genes. The present findings indicate that early-in life dietary zinc deficiency or supplementation did not significantly modify the susceptibility to development of mammary tumors induced by DMBA. Overall design: Pregnant Sprague-Dawley (SD) rats (16 dams /group) were distributed into three groups: ZnA, dams fed adequate zinc diet (35 mg/Kg chow); ZnD, dams fed zinc deficient diet (3 mg/Kg chow); and ZnS, dams fed zinc supplemented diet (180 mg/Kg chow). These experimental diets were provided ad libitum to the dams from GD 10 until lactational day (LD) 21 (weaning). At PND 21, female offspring were allocated in their respective dietary intervention and received the same diet that dams intake during gestation and lactation phases until PND 51 (puberty). At PND 51, all animals received a single intragastric administration (i.g.) of DMBA (50 mg/kg, diluted in canole oil) and were fed adequate zinc diet until PND 180. At PND 180 the animals were euthanized for mammary tumor and gene expression analyses.
Project description:Developmental programming is the concept that environmental factors, particularly during foetal life, can alter development, metabolism and physiology of an organism and this can have consequences later in life. There is growing interest in developmental programming in livestock species, particularly effects of maternal pregnancy nutrition, which is easy to manipulate. Recent research, using a sheep model, has shown that milk production in ewe offspring may be susceptible to maternal nutritional programming, such that over nutrition (ad libitum) of the pregnant dam, compared with maintenance nutrition, may impair their first lactation performance and result in the weaning of lighter lambs. RNA-seq was performed to identify gene expression differences as a result of maternal nutrition in ewe offspring during their first parity. Samples were collected in late pregnancy and during lactation, allowing us to examine gene expression changes during maturation of the ovine mammary gland. Three biological replicates were sequenced for each of the treatment conditions (maternal nutrition: sub-maintenance, maintenance, and ad libitum) and time points (late pregnancy and lactation). Each biological replicate consisted of RNA from multiple individuals (late pregnancy n=3, lactation n=2).
Project description:Background: Consumption of high fat diets has negative impacts on health and well-being, some of which may be epigenetically regulated. Selenium and folate are two compounds which influence epigenetic mechanisms. We investigated the hypothesis that post-weaning supplementation with adequate levels of selenium and folate in mouse offspring fed a high fat, low selenium and folate diet during gestation and lactation will lead to epigenetic changes of potential importance for long-term health. Female offspring of mothers fed the experimental diet were either maintained on this diet (HF-low-low), or weaned onto a high-fat diet with sufficient levels of selenium and folate (HF-low-suf), for 8 weeks. Gene and protein expression, DNA methylation, and histone modifications were measured in colon and liver of female offspring. Results: Adequate levels of selenium and folate post-weaning affected gene expression in colon and liver of offspring, including decreasing Slc2a4 gene expression. Protein expression was only altered in the liver. There was no effect of adequate levels of selenium and folate on global histone modifications in the liver. Global liver DNA methylation was decreased in mice switched to adequate levels of selenium and folate, but there was no effect on methylation of specific CpG sites within the Slc2a4 gene in liver. Conclusions: Post-weaning supplementation with adequate levels of selenium and folate in female offspring of mice fed high-fat diets during gestation and lactation can alter global DNA methylation in liver. This may be one mechanism by which the negative effects of a poor diet during early life can be ameliorated. Further research is required to establish what role epigenetic changes play in mediating observed changes in gene and protein expression, and the relevance of these changes to health. Female wild type C57BL/6 mice (Animal Resource Centre, Western Australia) were fed a High Fat diet containing low levels of selenium and folate (HF-Low) for 7 days prior to mating with male C57BL/6 mice (Ruakura Small Animal Facility, Hamilton, New Zealand). Mothers were maintained on the HF-Low diet throughout gestation and lactation. Offspring of these female mice were randomly assigned to one of two different dietary treatments: either the same diet as the mothers (HF-Low), or a High Fat diet containing adequate selenium and folate (HF-Suf). At 12 weeks of age, mice were euthanized and colon and liver samples taken for microarray, proteomics, and DNA methylation analyses. Genomic DNA, total RNA and protein from whole colon and liver tissue was extracted using an AllPrep® DNA/RNA/Protein mini kit (Qiagen, Cat number 80004). Colon and liver RNA from six female offspring on the HF-Low diet was compared with colon and liver RNA from six female offspring on the HF-Suf diet. All individual RNA samples were hybridized against a common reference RNA on separate arrays. The reference RNA was prepared by pooling in equimolar proportions RNA extracted from the intestine and liver of twelve female C57BL/6 mice, these being all of the mice from which samples were derived for microarray analysis in the current study.