Fat and carbohydrate intake over three generations modify growth, metabolism and cardiovascular phenotype in female mice in an age-related manner

Environmental challenges such as a high fat diet during pregnancy can induce changes in offspring growth, metabolism and cardiovascular function. However, challenges that are sustained over several generations can induce progressive compensatory metabolic adjustments in young adults. It is not known if such effects persist during ageing. We investigated whether diets with different fat and carbohydrate contents over three generations modifies markers of ageing. Female C57BL/6 F0 mice were fed diets containing 5% or 21% fat (w/w) throughout pregnancy and lactation. Female offspring were fed the same diet as their dams until the F3 generation. In each generation, body weight, 24-hour food intake were recorded weekly, and plasma metabolites were measured by colorimetric assays, blood pressure by tail cuff plethysmography and vasoconstriction by myography on postnatal day 90 or 456. There was little effect of diet or generation on phenotypic markers in day 90 adults. There was a significant increase in whole body, liver and heart weight with ageing (d456) in the F3 21% fat group compared to the F1 and F3 5% groups. Fasting plasma glucose concentration was significantly increased with ageing in the 5% group in the F3 generation and in the 21% group in both generations. There was a significant effect of diet and generation on ex-vivo vasoconstriction in ageing females. Differences in dietary fat may induce metabolic compensation in young adults that persist over three generations. However, such compensatory effects decline during ageing

Progressive, Transgenerational Changes in Offspring Phenotype and Epigenotype following Nutritional Transition

Induction of altered phenotypes during development in response to environmental input involves epigenetic changes. Phenotypic traits can be passed between generations by a variety of mechanisms, including direct transmission of epigenetic states or by induction of epigenetic marks de novo in each generation. To distinguish between these possibilities we measured epigenetic marks over four generations in rats exposed to a sustained environmental challenge. Dietary energy was increased by 25% at conception in F0 female rats and maintained at this level to generation F3. F0 dams showed higher pregnancy weight gain, but lower weight gain and food intake during lactation than F1 and F2 dams. On gestational day 8, fasting plasma glucose concentration was higher and β-hydroxybutyrate lower in F0 and F1 dams than F2 dams. This was accompanied by decreased phosphoenolpyruvate carboxykinase (PEPCK) and increased PPARα and carnitine palmitoyl transferase-1 mRNA expression. PEPCK mRNA expression was inversely related to the methylation of specific CpG dinucleotides in its promoter. DNA methyltransferase (Dnmt) 3a2, but not Dnmt1 or Dnmt3b, expression increased and methylation of its promoter decreased from F1 to F3 generations. These data suggest that the regulation of energy metabolism during pregnancy and lactation within a generation is influenced by the maternal phenotype in the preceding generation and the environment during the current pregnancy. The transgenerational effects on phenotype were associated with altered DNA methylation of specific genes in a manner consistent with induction de novo of epigenetic marks in each generation

Vascular Dysfunction Induced in Offspring by Maternal Dietary Fat Involves Altered Arterial Polyunsaturated Fatty Acid Biosynthesis

Nutrition during development affects risk of future cardiovascular disease. Relatively little is known about whether the amount and type of fat in the maternal diet affect vascular function in the offspring. To investigate this, pregnant and lactating rats were fed either 7%(w/w) or 21%(w/w) fat enriched in either18:2n-6, trans fatty acids, saturated fatty acids, or fish oil. Their offspring were fed 4%(w/w) soybean oil from weaning until day 77. Type and amount of maternal dietary fat altered acetylcholine (ACh)-mediated vaso-relaxation in offspring aortae and mesenteric arteries, contingent on sex. Amount, but not type, of maternal dietary fat altered phenylephrine (Pe)-induced vasoconstriction in these arteries. Maternal 21% fat diet decreased 20:4n-6 concentration in offspring aortae. We investigated the role of Δ6 and Δ5 desaturases, showing that their inhibition in aortae and mesenteric arteries reduced vasoconstriction, but not vaso-relaxation, and the synthesis of specific pro-constriction eicosanoids. Removal of the aortic endothelium did not alter the effect of inhibition of Δ6 and Δ5 desaturases on Pe-mediated vasoconstriction. Thus arterial smooth muscle 20:4n-6 biosynthesis de novo appears to be important for Pe-mediated vasoconstriction. Next we studied genes encoding these desaturases, finding that maternal 21% fat reduced Fads2 mRNA expression and increased Fads1 in offspring aortae, indicating dysregulation of 20:4n-6 biosynthesis. Methylation at CpG −394 bp 5′ to the Fads2 transcription start site predicted its expression. This locus was hypermethylated in offspring of dams fed 21% fat. Pe treatment of aortae for 10 minutes increased Fads2, but not Fads1, mRNA expression (76%; P<0.05). This suggests that Fads2 may be an immediate early gene in the response of aortae to Pe. Thus both amount and type of maternal dietary fat induce altered regulation of vascular tone in offspring though differential effects on vaso-relaxation, and persistent changes in vasoconstriction via epigenetic processes controlling arterial polyunsaturated fatty acid biosynthesis

Supplementation with n-3 long-chain polyunsaturated fatty acids or olive oil in men and women with renal disease induces differential changes in the DNA methylation of FADS2 and ELOVL5 in peripheral blood mononuclear cells

BACKGROUND: Studies in animal models and in cultured cells have shown that fatty acids can induce alterations in the DNA methylation of specific genes. There have been no studies of the effects of fatty acid supplementation on the epigenetic regulation of genes in adult humans.METHODS AND RESULTS: We investigated the effect of supplementing renal patients with 4 g daily of either n-3 long-chain polyunsaturated fatty acids (n-3 LCPUFA) or olive oil (OO) for 8 weeks on the methylation status of individual CpG loci in the 5' regulatory region of genes involved in PUFA biosynthesis in peripheral blood mononuclear cells from men and women (aged 53 to 63 years). OO and n-3 LCPUFA each altered (&gt;10% difference in methylation) 2/22 fatty acid desaturase (FADS)-2 CpGs, while n-3 LCPUFA, but not OO, altered (&gt;10%) 1/12 ELOVL5 CpGs in men. OO altered (&gt;6%) 8/22 FADS2 CpGs and (&gt;3%) 3/12 elongase (ELOVL)-5 CpGs, while n-3 LCPUFA altered (&gt;5%) 3/22 FADS2 CpGs and 2/12 (&gt;3%) ELOVL5 CpGs in women. FADS1 or ELOVL2 methylation was unchanged. The n-3 PUFA supplementation findings were replicated in blood DNA from healthy adults (aged 23 to 30 years). The methylation status of the altered CpGs in FADS2 and ELOVL5 was associated negatively with the level of their transcripts.CONCLUSIONS: These findings show that modest fatty acid supplementation can induce altered methylation of specific CpG loci in adult humans, contingent on the nature of the supplement and on sex. This has implications for understanding the effect of fatty acids on PUFA metabolism and cell function

Different dietary omega-3 sources during pregnancy and DHA in the developing rat brain

The essential n-3 fatty acid α-linolenic acid (ALA) can be converted into eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) under the action of desaturase and elongase enzymes. Human studies have demonstrated that females convert a higher proportion of ALA into EPA and DHA than males. We have demonstrated that when fed upon an ALA rich diet, female rats have a significantly higher EPA content of plasma and liver lipids than males. When fetal tissues were collected, it was observed that pups from dams fed the ALA rich diet had a comparable brain DHA status to those from dams fed on a salmon-oil based diet, indicating that conversion of ALA to DHA during pregnancy was efficient, and that DHA accumulated in a tissue-specific manner. Similar efficacy of dietary ALA in women during pregnancy would mean that plant n-3 fatty acids would be useful alternatives to preformed EPA and DHA

Different dietary omega-3 sources during pregnancy and DHA in the developing rat brain

The essential n-3 fatty acid α-linolenic acid (ALA) can be converted into eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) under the action of desaturase and elongase enzymes. Human studies have demonstrated that females convert a higher proportion of ALA into EPA and DHA than males. We have demonstrated that when fed upon an ALA rich diet, female rats have a significantly higher EPA content of plasma and liver lipids than males. When fetal tissues were collected, it was observed that pups from dams fed the ALA rich diet had a comparable brain DHA status to those from dams fed on a salmon-oil based diet, indicating that conversion of ALA to DHA during pregnancy was efficient, and that DHA accumulated in a tissue-specific manner. Similar efficacy of dietary ALA in women during pregnancy would mean that plant n-3 fatty acids would be useful alternatives to preformed EPA and DHA

Maternal diet during pregnancy has tissue-specific effects upon fetal fatty acid composition and alters fetal immune parameters

Both animalandhumanstudiesdemonstratethatthedocosahexaenoicacid(DHA)contentofplasmaand/ortissuelipidsisincreasedduringpregnancy.Wehypothesisedthatincreasingthe a-linolenicacid(ALA)orlongerchain(n-3) PUFAcontentofthematernaldietduringpregnancyinfluencesfetalfattyacid compositionandthefetalimmunesystem.Pregnantratswerefedalow-fat(LF)soybeanoildiet,orhigh-fat(HF)soybean,linseed,salmonorsunfloweroildietsfromconceptionto20dgestation.TheALA-richLinseed-HFdietresultedinanequivalenteicosapentaenoicacid(EPA)statusinfetalimmunetissuesandanequivalentDHAstatusinthefetalbraintothatachievedwiththeSalmon-HFdiet.An(n-3) richmaternaldietduringpregnancyassociatedwiththehighestexpressionofCD3(Salmon-HF)and CD8(Linseed-HFandSalmon-HF)onfetalthymicCD3+CD8+ cells. TheLinseed-HFdietresultedinthe highestproportionofCD161+ cells withinthefetalthymus,whichcorrelatedwiththeproductionofIL-4. ThesedataindicatethatdietaryALAsupplementationmayconfersomeofthebenefitsofLC(n-3)PUFA duringpregnancy.Thisshouldbeexaminedinsuitablydesignedhumanstudie

Correction: Fat and Carbohydrate Intake over Three Generations Modify Growth, Metabolism and Cardiovascular Phenotype in Female Mice in an Age-Related Manner.

[This corrects the article DOI: 10.1371/journal.pone.0134664.]