S100 Posters

the TFRC S142G polymorphism (190.4g, p = 0.03). No statistically

significant effect was seen in female infants.

Conclusion: Polymorphisms in the HFE and TFRC genes are

associated with significantly elevated infant birth weight in males.

This observation suggests that the HFE H63D, C282Y and TFRC

S142G variants are important determinants of fetal growth. Further

investigation of these variants and the role of iron metabolism in

susceptibility to disease in later life is warranted.

The authors acknowledge the North of England Children’s Cancer

Research Fund for funding.

P1-63 DNA methylation status at the Igf2 locus in offspring of

a mouse model of maternal diet-induced obesity

A. Groom1, C.L. Relton1 *, J.A. McKay1, L. Steedman1,

J.C. Mathers1, P. Taylor2, L. Poston2. 1Human Nutrition Research

Centre, Newcastle University, UK, 2King’s College London, London,

UK

E-mail: c.l.relton@ncl.ac.uk

Aims: Chronic over-nutrition during pregnancy results in phenotypic

changes in offspring, as demonstrated in the maternal diet-induced

obesity mouse model. Neonates go on to develop hyperphagia,

obesity (increased abdominal adiposity) with subsequent hyperten-

sion, dyslipidaemia and insulin resistance from around 3 months of

age. This pilot study investigated the postulated role of epigenetic

mechanisms in mediating nutritional programming by analysing DNA

methylation at the Igf2 locus.

Study design: Female C57Bl/6 mice were fed a control diet

(RM 1.5% corn oil) or a highly palatable diet (OB2 21% lard

and sweetened condensed milk) prior to mating and throughout

pregnancy and lactation. Offspring were weaned to standard chow.

DNA was extracted from liver tissue from 3 month old female

offspring for methylation assessment (n = 6 per group).

Outcome measures: Percentage DNA methylation of 13 CpG sites

within the differentially methylated region (DMR2) of the Igf2 gene

was measured by pyrosequencing following bisulphite conversion of

DNA.

Results: High levels of reproducibility were observed between du-

plicate samples and replicate assays when determining methylation

levels by Pyrosequencing. Subtle elevation in DNA methylation was

observed at 8/13 CpG sites at the Igf2 DMR2 site.

Conclusion: Preliminary data suggests that subtle but statistically

non-significant differential methylation is observed at the Igf2

DMR2 site in liver tissues from 3 month old offspring of diet-induced

obese mice when compared to offspring of control mice.

The authors acknowledge the BBSRC (CISBAN), EARNEST (EUFP6)

and NuGO (EU FP6) for funding.

P1-64 Epigenetic regulation of the HNF4a (MODY 1) gene: role

in developmental programming of type 2 diabetes

I. Sandovici1 *, N.H. Smith2, M. Constancia1, S.E. Ozanne2. 1The

Babraham Institute, Cambridge, UK, 2Department of Clinical

Biochemistry, University of Cambridge, UK

E-mail: ionel.sandovici@bbsrc.ac.uk

Aims: This study investigated the role of epigenetic-induced

changes in expression of HNF4a in pancreatic islets in mediating

the relationship between poor early growth and type 2 diabetes.

Study design: The protein restriction rat model was used to

determine if nutrition in early life induced changes in HNF4a, an

important developmental transcription factor, mutations in which

cause diabetes.

Subjects: Two groups of offspring were studied (n = 10). Control and

low protein animals were the offspring of dams fed a 20% protein

diet or an isocaloric 8% protein diet, respectively, during pregnancy

and lactation. All animals were weaned onto standard chow and

studied at 3 months of age.

Outcome measures: Gene expression was determined by Q-PCR

and western blotting, DNA methylation by bisulphite sequencing

and histone modifications by chromatin immunoprecipitation.

Results: HNF4a expression was reduced by 40% at mRNA level

and by 80% at protein level in low protein offspring. This was

accompanied by reductions in downstream targets of HNF4a:

Kir6.2, Glut2 and l-Pk. We found tissue-specific methylation

differences at the promoter region of HNF4a that correlated with

gene activity. The main active promoter in pancreatic islets is

unmethylated, but was hypermethylated and silent in kidney of

control animals. We are currently profiling histone modifications

and investigating how epigenetic marking diversity contributes to

programming effects at this locus.

Conclusions: Hnf4a is a target of nutritional programming, linking

a suboptimal early environment to increased diabetes risk. DNA

methylation may play an important role in regulating the activity

of Hnf4a promoters in a tissue-specific manner.

P1-65 Periconceptional undernutrition (PCUN) and

twinning in sheep differently alter the ontogeny of

adrenocorticotrophin receptor (ACTH-R), steroidogenic

acute regulatory protein (StAR) and P450C17 (CYP17) in

fetal adrenal glands

F.H. Bloomfield1,2 *, K.L. Connor3, A.L. Jaquiery1,2, H.H. Phua1,

M.H. Oliver1,2, J.R.G. Challis3,4, J.E. Harding1,2. 1Liggins Institute,

University of Auckland, Auckland, New Zealand, 2National

Research Centre for Growth and Development, New Zealand,3Department of Physiology, and 4Departments of Obstetrics and

Gynaecology and of Medicine, University of Toronto, Toronto,

Canada

E-mail: f.bloomfield@auckland.ac.nz

Aims: to determine the effects of PCUN and twinning on ontogeny

of the ACTH-R, StAR and CYP17 in the fetal adrenal gland.

Study design: Ewes were randomly assigned to ad libitum feed

throughout (N), or PCUN from 60d before to 30d after mating,

tailored to reduce maternal weight by 15%, with ad libitum

thereafter. At 50, 85, 120 and 131d (term = 147d) mRNA levels

in fetal adrenal glands were determined by real-time RT-PCR.

Outcome measures: Data are presented as percent change of the

relative expression ratio of target gene to 18s rRNA.

Results: There were no significant effects of PCUN or twinning

on ACTH-R, StAR or CYP17 at 50d gestation. PCUN in singletons

resulted in increases of: 80% in CYP17 by d85; 850% in CYP17 and

700% in StAR at 120d; and of 560% in CYP17 and 350% in StAR

at 131d. PCUN in twins resulted in similar, but less strong, changes.

Twinning, in N animals, did not result in any significant differences.

Twinning, in PCUN animals, increased CYP17 120% and StAR 130%

at 85d compared with PCUN singletons, but profoundly suppressed

CYP17 by 80% at 120d and suppressed StAR by 40% and CYP17 by

50% at 131d.

Conclusions: Both PCUN and twinning, different periconceptional

events, result in early increases in CYP17 mRNA levels. However,

whereas in PCUN CYP17 increases further in late gestation, in

twinning CYP17 mRNA levels are profoundly suppressed. The

mechanism behind this switch may explain why PCUN fetuses

deliver early, but twin fetal sheep do not.

P1-66 Impact of genotype and fetal and pre-weaning growth

on steroidogenic capacity of the adult bovine adrenal

C.L. Coulter1 *, P.L. Greenwood2, S.L. Dunn1, M.D. Salkeld1.1School of Molecular & Biomedical Sciences, University of

Adelaide; 2NSW DPI, University of New England, Armidale,

Australia

E-mail: ccoulter@u.washington.edu

Background: Recent studies have suggested a role of the

hypothalamic-pituitary-adrenal axis in the epidemiological associ-

ation between birth weight and cardiovascular disease.

Aims and Study design: The aims of this study were to determine

if the steroidogenic capacity of the adult adrenal gland is

permanently programmed by exposure to either high or low

nutrition and growth before birth followed by either high or low