The Journal of Nutrition
Biochemical, Molecular, and Genetic Mechanisms
Folic Acid Supplementation during the
Juvenile-Pubertal Period in Rats Modifies
the Phenotype and Epigenotype Induced by
Graham C. Burdge,4* Karen A. Lillycrop,5Emma S. Phillips,6Joanne L. Slater-Jefferies,6Alan A. Jackson,4
and Mark A. Hanson6
4Institute of Human Nutrition,5Development and Cell Biology, and6Institute of Developmental Sciences, University of Southampton,
Southampton SO16 6YD, UK
Prenatal nutritional constraint is associated with increased risk of metabolic dysregulation in adulthood contingent on adult
diet. In rats, folic acid supplementation of a protein-restricted (PR) diet during pregnancy prevents altered phenotype and
epigenotype in the offspring induced by the PR diet. We hypothesized that increasing folic acid intake during the juvenile-
pubertal (JP) period would reverse the effects of a maternal PR diet on the offspring. Rats were fed a control (C) or PR diet
during pregnancy and AIN93G during lactation. Offspring were weaned on d 28 onto diets containing 1 mg [adequate
folate (AF)] or 5 mg [folic acid-supplemented (FS)] folic acid/kg feed. After 28 d, all offspring were fed a high-fat (18% wt:
wt) diet and killed on d 84. As expected, offspring of PR dams fed the AF diet had increased fasting plasma triglyceride
(TAG) and b-hydroxybutyrate (bHB) concentrations. The FS diet induced increased weight gain, a lower plasma bHB
concentration, and increased hepatic and plasma TAG concentration compared with AF offspring irrespective of maternal
diet. PPARa andglucocorticoid receptorpromotermethylationincreasedin liverand insulinreceptorpromotermethylation
decreased in liver and adipose tissue in FS compared with AF offspring, with reciprocal changes in mRNA expression
the phenotype induced by the maternal diet. This may represent a period of plasticity when specific nutrient intakes may
alter the phenotype of the offspring through epigenetic changes in specific genes.J. Nutr. 139: 1054–1060, 2009.
Environmental constraint during development, including under-
nutrition, is causally associated with increased risk of metabolic
disease (1). It has been suggested that the developing fetus
responds to environmental cues via developmental plasticity
in a manner that sets the phenotype in prediction of the future
environment, conferring a Darwinian fitness advantage (2). In
humans, mismatch between the predicted and future environ-
ments has been suggested to underlie the early life origins of
metabolic disease (2). Several animal models show that subop-
timal nutrition during critical periods in early development
induces metabolic dysregulation that resembles human cardio-
metabolic disease, especially if the offspring are fed a high-fat
diet (3,4). In humans and in animal models, the nature of the
phenotype induced by maternal dietary constraint differs
according the developmental stage of the fetus at the time of
exposure (5,6). Thus, the response of individual tissues to
nutrient constraint is dependent upon the stage of maturation at
the time of exposure.
Moderate maternal dietary protein restriction in rats is a
well-established model of induction of an altered phenotype in
the offspring (7). The offspring are characterized by persistent
hypertension (7,8), dyslipidemia, and impaired glucose metab-
olism (4). Supplementation of the maternal protein-restricted
(PR)7diet with folic acid or glycine prevents induction of
1Supported by a fellowship from the British Heart Foundation to G. C. B.
(FS/05/064/19525). M. A. H. also receives salary support from the British Heart
2Author disclosures: G. C. Burdge, K. A. Lillycrop, E. S. Phillips, J. L.
Slater-Jefferies, A. A. Jackson, and M. A. Hanson, no conflicts of interest.
3Supplemental Tables 1 and 2, and Supplemental Figure 1 are available with the
online posting of this paper at jn.nutrition.org.
*To whom correspondence should be addressed. E-mail: g.c.burdge@
7Abbreviations used: AF, folic acid-adequate; AOX, acyl-CoA carboxylase; bHB,
control/adequate folate; C/FS, control/folate supplemented; Dnmt, DNA methyl-
transferase; FS, folic acid-supplemented; GR, glucocorticoid receptor; HSL,
lipase; NEFA, nonesterified fatty acid; PEPCK, phosphoenolpyruvate carboxyki-
nase; pn, postnatal; PR, protein-restricted, PR/AF, protein-restricted/adequate
folate; PR/FS, protein-restricted/folate supplemented; TAG, triglyceride.
0022-3166/08 $8.00 ã 2009 American Society for Nutrition.
Manuscript received January 14, 2009. Initial review completed February 24, 2009. Revision accepted March 5, 2009.
First published online April 1, 2009; doi:10.3945/jn.109.104653.
by guest on December 26, 2015
Supplemental Material can be found at:
fitness by predicting the future environment and that an incorrect
prediction is associated with increased risk of later chronic
disease (2). Our findings suggest this hypothesis could be
extended to include the phenotypic changes induced during the
subsequent period of plasticity in puberty, perhaps enhancing
the organism’s life course strategy.
We thank John Jackson, Samuel Hoile, Rebecca Grant, and the
staff of the Biomedical Research Facility, University of South-
ampton, for technical assistance.
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