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Adam J Watkins Centre for Biological Sciences, School of Life and Health Sciences, Faculty of Life Sciences, Southampton General Hospital, University of Southampton, Southampton SO16 6YD, UK
Centre for Biological Sciences, School of Life and Health Sciences, Faculty of Life Sciences, Southampton General Hospital, University of Southampton, Southampton SO16 6YD, UK

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Emma S Lucas Centre for Biological Sciences, School of Life and Health Sciences, Faculty of Life Sciences, Southampton General Hospital, University of Southampton, Southampton SO16 6YD, UK

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Stephanie Marfy-Smith Centre for Biological Sciences, School of Life and Health Sciences, Faculty of Life Sciences, Southampton General Hospital, University of Southampton, Southampton SO16 6YD, UK

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Nicola Bates Centre for Biological Sciences, School of Life and Health Sciences, Faculty of Life Sciences, Southampton General Hospital, University of Southampton, Southampton SO16 6YD, UK

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Susan J Kimber Centre for Biological Sciences, School of Life and Health Sciences, Faculty of Life Sciences, Southampton General Hospital, University of Southampton, Southampton SO16 6YD, UK

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Tom P Fleming Centre for Biological Sciences, School of Life and Health Sciences, Faculty of Life Sciences, Southampton General Hospital, University of Southampton, Southampton SO16 6YD, UK

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Mammalian placentation is dependent upon the action of trophoblast cells at the time of implantation. Appropriate fetal growth, regulated by maternal nutrition and nutrient transport across the placenta, is a critical factor for adult offspring long-term health. We have demonstrated that a mouse maternal low-protein diet (LPD) fed exclusively during preimplantation development (Emb-LPD) increases offspring growth but programmes adult cardiovascular and metabolic disease. In this study, we investigate the impact of maternal nutrition on post-implantation trophoblast phenotype and fetal growth. Ectoplacental cone explants were isolated at day 8 of gestation from female mice fed either normal protein diet (NPD: 18% casein), LPD (9% casein) or Emb-LPD and cultured in vitro. We observed enhanced spreading and cell division within proliferative and secondary trophoblast giant cells (TGCs) emerging from explants isolated from LPD-fed females when compared with NPD and Emb-LPD explants after 24 and 48 h. Moreover, both LPD and Emb-LPD explants showed substantial expansion of TGC area during 24–48 h, not observed in NPD. No difference in invasive capacity was observed between treatments using Matrigel transwell migration assays. At day 17 of gestation, LPD- and Emb-LPD-fed conceptuses displayed smaller placentas and larger fetuses respectively, resulting in increased fetal:placental ratios in both groups compared with NPD conceptuses. Analysis of placental and yolk sac nutrient signalling within the mammalian target of rapamycin complex 1 pathway revealed similar levels of total and phosphorylated downstream targets across groups. These data demonstrate that early post-implantation embryos modify trophoblast phenotype to regulate fetal growth under conditions of poor maternal nutrition.

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