LAUSR

A novel, non-terminal surgical procedure to remove a single placentome from the pregnant ewe for gene expression and histological analyses was recently developed in our laboratory. This technique allows for evaluation of nutritional insults on placental development at more than one stage of gestation using a single animal. Early attempts to develop a similar technique in cattle were met with complications due to inaccessibility of the gravid uterine horn because of its location and mass. One alternative is to collect a placentome from the contralateral uterine horn: however, the question remains as to whether gene expression varies among placentomes based on location relative to the fetus. Pregnant heifers were maintained on forage during early gestation and later moved into pens with a Calan gate system (American Calan, Northwood, NH). On gestational day (GD) 158, five heifers were assigned to receive a hay-based diet formulated to meet 100% of NRC requirements, and five heifers were fed 70% of NRC requirements until necropsy on GD 244. At necropsy, a single representative placentome was selected for analysis from the antimesometrial side: 1) of the gravid uterine horn central to the amnion, 2) over the allantois immediately adjacent to the amnion, 3) in the tip of the gravid uterine horn, and 4) in the tip of the contralateral uterine horn. Mean placentome weight was greater (P<0.05) for locations central to the amnion and allantois compared to locations within the tips of the ipsilateral and contralateral horns, respectively. Gene expression for angiogenic factors (FGF2, ODC1, VEGFA,FLT1), nutrient transporters (SLC7A1, SLC2A1), and factors associated with hormone action (ESR1, IGF1, IGFBP3, CSH1,PAG1) were unaffected (P>0.05) by dietary treatment or location of the placentome. Results indicate that location of the placentome in relation to the fetus does not impact gene expression, enhancing the efficacy of non-terminal methodologies for sampling gene expression in placentomes.