a periconceptional maternal hyperglycemia disrupts the feto-placental membrane fatty acid profiles...

Abstracts / Placenta 35 (2014) A1eA112 A29

centrifugation. The total number of exosomes and PdEwere determined byquantifying immunoreactive exosomal CD63 and placenta-specific marker(PLAP) by ELISA and fluorescence nanoparticle tracking analysis (NTA,NanoSight NS500). The PLAP/CD63 ratio (i.e. immunoractive PLAP contentper exosome) was used as a measure of the contribution of PdE to totalexosomes in maternal blood.Results: Variation in the concentration of PdE in maternal plasma wasassessed by ANOVAwith the variance partitioned between gestational ageand pregnancy status (i.e. normal or GDM). Both gestational age andpregnancy status were identified as significant factors (ANOVA, p<0.05).Post-host analyses established that PdE concentrations increased duringgestation (FT, ST and TT) in both normal and GDM pregnancies, however,the increase was significantly greater in GDM. (~2.2-fold, ~1.5-fold and~1.8-fold greater in the FT, ST and TT compared to normal pregnancies,respectively.) We observed that PLAP ratio was decreased dramatically inGDM pregnancies (p < 0.05).Conclusions: The concentrations of exosomes in maternal blood arehigher in GDM than normal pregnancies. These characteristics couldpotentially be used for diagnostic markers for exosome profiling to screenasymptomatic populations.

P1.60.A PERICONCEPTIONAL MATERNAL HYPERGLYCEMIA DISRUPTS THEFETO-PLACENTAL MEMBRANE FATTY ACID PROFILES IN A RABBITMODEL

Delphine Rousseau-Ralliard a, Emilie Derisoud a, Anne Tarrade a, RoselyneBrat a, Audrey Rolland a, Rene Thieme b, Anne Navarette-Santos b, BerndFischer b, Pascale Chavatte-Palmer a a INRA, UMR 1198 Biology ofDevelopment and Reproduction, Jouy-en-Josas, France; bMLU, Faculty ofMedicine, Department of Anatomy and Cell Biology, Halle, Germany

Type-1 diabetes (T1D) is caused by the reduction in pancreatic insulinsecretion, inducing chronic hyperglycemia. Pre-gestational T1D increasesthe risk of miscarriage and congenital malformations and programs theoffspring to develop metabolic syndrome at adulthood. Management ofmaternal diabetes is essential during the gestation but could be highlyimportant around the conception.Objectives: The aim of this study was to explore the effects of maternalTD1 during the periconceptional period on placenta and fetal phenotype at28dpc (term¼31days).Methods: Diabetes was induced by Alloxan in dams 7 days before mating.Glycemia was maintained at 15-20mmol/L with exogenous insulin in-jections. At 4dpc, embryos were collected and transferred into non-dia-betic recipients. At 28 dpc, control (C) and diabetic (D) fetuses werecollected for biometric records and lipid analyses of feto-placental tissuesby gas chromatography. The complex fatty acid profiles were analyzed byprincipal component analyses (PCA).Result: D-fetuses were growth retarded, hyperglycemic and dyslipidemiccompared to C. A specific fatty acid signature was observed in fetal plasma,with wel separated groups by PCA. The composition of placental and fetalliver membranes differed according to maternal status and fetal sex. Tis-sues from D-fetuses contained significantly more omega-6 poly-unsaturated fatty acids compared to C. No biochemical signature wasobserved in the immature fetal heart, but docosahexaenoic acid wasdecreased while linoleic acid increased in the cardiac membranes of D-fetuses, indicating a higher risk of ischemia.Conclusion: This study demonstrates that an exposure to high plasmaglucose during the short periconceptional period reduces fetal growth andalters the lipid profiles of placenta and fetal tissues.

P1.61-N.INVESTIGATING FATTY ACID TRANSPORT AND b- FATTY ACIDOXIDATION (FAO) IN PLACENTAS EXPOSED TO HYPERGLYCEMIA

Charlotte Hulme a,b, Melissa Westwood a,b, Alexander EP. Heazell a,b, JennyMyers a,b aMaternal and Fetal Health Research Centre, Manchester, UK;bManchester Academic Health Sciences Centre, University of Manchester, UK

Objective: Pregnancy complications associated with Diabetes Mellitus(DM) are likely to result from placental dysfunction. Previous studies of

gestational DM show that in hyperglycemia, malonyl-CoA, produced in areaction catalysed by acetyl-CoA carboxylase (ACC1), inhibits placentalfatty acid oxidation (FAO) leading to increased triglyceride levels. Networkanalysis of ‘omic data from BeWo cells cultured in different glucose con-centrations suggested consequent alterations in lipid metabolism. There-fore, this study aimed to characterise fatty acid transport and oxidation interm placental explants exposed to high glucose levels in vitro anddetermine whether any detrimental effects might be countered by stra-tegies to manipulate FAO.

Methods:Normal placental villous explants (n¼6) were cultured for 18h inDMEM:F12 containing 5mM or 25mM D-glucose, with and without anACC1 inhibitor (5-(Tetradecyloxy)-2-furoic acid (TOFA); 5mg/ml) or agonistof PPARa, (Clofibrate; 10-40mM), which up-regulates FAO enzymes, for afurther 3 or 24h. Expression of the fatty acid transporters, fatty acidtranslocase, fatty acid transport protein (FATP) 2 and FATP4 was assessedusing immunohistochemistry. The level of triglyceride in cultured explantswas measured using a commercial assay (Caymann).Result: Placental explants cultured with 25mM glucose had significantlygreater triglyceride levels. Addition of TOFA, but not Clofibrate, reversedthe effects of high glucose, reducing triglycerides to levels similar to thosemeasured in explants cultured in 5mM D-glucose (p<0.05). Culture in ahigh glucose environment did not alter trophoblast, endothelial or stromalexpression of fatty acid transporters (p>0.05).Conclusion: Fatty acid transporter expression is unaltered in placentaltissue exposed to high glucose in vitro. These data suggest that maternalhyperglycemia might affect placental fatty acid metabolism rather thantransfer across the placenta. Indeed glucose-induced attenuation of FAOcan be reversed by inhibition of ACC1. Subsequent studies will explorewhether this pathway is altered in placentas from pregnancies compli-cated by type1/2 DM.

P1.62.SOLUBLE PLACENTAL FRACTION SIGNALS VIA CHEMOKINE RECEPTOR 1AND 3 ON TROPHOBLAST STEM-CELL-LIKE POPULATION FOR RAPIDAND GUIDED MIGRATION TO DISTRESSED AREAS

Geraldine Gascoin a,b, Denise Fernandez-Twinn a, Daniella Duque a, SarahCarr a, Barbara Musial a, Heather Blackmore a, Suzan Ozanne a a Instituteof Metabolic Sciences, University of Cambridge, Cambridge, UK;bDepartment of Neonatal Medicine, Angers University Hospital, Angers,France

Background: Overweight or obesity prevalence in pregnant women hasincreased in line with the global obesity epidemic: up to 50% of womenof reproductive age and 20-25% of pregnant women at first antenatalvisits in Europe and the USA. Epidemiological and animal data suggestthat maternal obesity during pregnancy adversely affects offspringhealth. The underlying mechanisms may involve maternal and fetaldysregulation of glucose, insulin, lipid and amino acid metabolism. Theplacenta develops to support fetal growth, and therefore plays a key rolein the aetiology of developmental programming by impacting onnutrient transfer.

Objective: The objective of this study was to analyse the effect of exerciseon placenta in obesogenic diet exposed mouse dams.Methods: The study consisted of 3 groups: control (n¼5), obese (n¼5) andobese + exercise (n¼5). The obesogenic diet was for 6 weeks before firstmating in the 2 obese groups. After weaning, the obese + exercise group,commenced training for a week (20 min/day, 5 days/week) before matingfor second pregnancy and exercised until day 17 of gestation. The twoother groups were also mated for second pregnancies. Placentas werecollected at day 19 for morphometry and staining and frozen for proteinand gene expression analyses.Results: Exercise intervention reduced placenta lipid storage and transferto the fetal trophoblast. It also normalised expression of some insulinsignalling components that were dysregulated in the placentas of unex-ercised obese dams.Conclusion: Exercise starting before and maintained during pregnancymay improve fetal outcome by restoring placental nutrient transfer.