Maternal nutrient restriction and over-feeding during gestation alter expression of key factors involved in placental development and vascularization

Poor maternal nutrition can negatively affect fetal and placental growth and development. However, the mechanism(s) that contribute to altered placenta growth and function are not well understood. We hypothesized that poor maternal diet would impact signaling through the C-X-C motif chemokine ligand (CXCL) 12-CXCL4 axis and/or placental expression of the insulin-like growth factor (IGF) axis. Using our established sheep model of poor maternal nutrition, we examined the effects of restricted- and over-feeding on ewe placentome gene and protein expression. Specifically, ewes were fed a control (CON; 100%), restricted (RES; 60%), or over (OVER; 140%) diet beginning at day 30.2 +/- 0.02 of gestation, and samples were collected at days 45, 90, and 135 of gestation, representing periods of active placentation, peak placental growth, and near term, respectively. Placentomes were separated into cotyledon and caruncle, and samples snap frozen. Protein was determined by western blot and mRNA expression by real-time PCR. Data were analyzed by ANOVA and significance determined at P <= 0.05. Ewes fed a RES diet had decreased CXCL12 and vascular endothelial growth factor (VEGF), and increased tumor necrosis factor (TNF)alpha protein compared with CON ewes in caruncle at day 45 (P <= 0.05). In day 45 cotyledon, CXCR7 protein was increased and mTOR was decreased in RES relative to CON (P <= 0.05). At day 90, CXCR4 and CXCR7 were reduced in RES caruncle compared with CON, whereas VEGF was reduced and mTOR increased in cotyledon of RES ewes relative to CON (P <= 0.05). In OVER caruncle, at day 45 CXCR4 and VEGF were reduced and at day 90 CXCR4, CXCR7, and TNF alpha were reduced in caruncle compared with CON (P <= 0.05). There was no observed effect of OVER diet on protein abundance in the cotyledon (P > 0.05). Expression of IGF-II mRNA was increased in OVER at day 45 and IGFBP-3 was reduced in RES at day 90 in caruncle relative to CON (P <= 0.05). Maternal diet did not alter placentome diameter or weight (P > 0.05). These findings suggest that restricted- and over-feeding negatively impact protein and mRNA expression of key chemokines and growth factors implicated in proper placenta development and function. Restricted and over-feeding during gestation negatively alter protein expression of key chemokines in the placenta and expression of growth factors that are required for placental vascularization. Lay Summary Too little or too much food during gestation can lead to poor growth and health of the resulting offspring. The placenta is an important source of nutrient supply for the fetus and poor maternal diet can impair placenta growth and function. Although placental development and function are well studied, the mechanisms by which maternal diet can affect placental growth and fetal development are not well understood. Based on our previous findings that specific proteins are important regulators of placental growth and function, we used a sheep model of poor maternal nutrition to demonstrate that protein abundance of these factors is altered in the placenta. These findings demonstrate potential mechanism by which maternal diet can affect the placenta and thereby impact fetal growth.