Miles Jeremy R, Farin Charlotte E, Rodriguez Karina F, Alexander Joseph E, Farin Peter W
Department of Population Health and Pathobiology, North Carolina State University, Raleigh, NC 27606-1499, USA.
Biol Reprod. 2004 Dec;71(6):1919-26. doi: 10.1095/biolreprod.104.031427. Epub 2004 Jul 30.
The objective of this study was to determine the effects of in vitro embryo production on angiogenesis and morphometry of the bovine placenta during late gestation. Blastocysts produced in vivo were recovered from superovulated Holstein cows. Blastocysts produced in vitro were obtained after culture of in vitro-matured and -fertilized Holstein oocytes. Single blastocysts from each production system were transferred into heifers. Fetuses and placentas were recovered on Day 222 of gestation (in vivo, n=12; in vitro, n=12). Cotyledonary and caruncular tissues were obtained for quantification of vascular endothelial growth factor (VEGF) and peroxisome proliferator-activated receptor-gamma (PPARgamma) mRNA and protein. Tissue sections of placentomes were prepared for morphometric analysis. Fetuses and placentas were heavier from embryos produced in vitro than from embryos produced in vivo. More placentas from embryos produced in vitro had an excessive volume of placental fluid. There was no effect of treatment on the expression of mRNA for VEGF and PPARgamma in either cotyledonary or caruncular tissues. The expression of VEGF protein in cotyledons and caruncles as well as the expression of PPARgamma protein in cotyledons were not different between the in vitro and in vivo groups. However, caruncles from the in vitro group had increased expression of PPARgamma protein. The total surface area of endometrium was greater for the in vitro group compared with controls. In contrast, the percentage placentome surface area was decreased in the in vitro group. Fetal villi and binucleate cell volume densities were decreased in placentomes from embryos produced in vitro. The proportional tissue volume of blood vessels in the maternal caruncles was increased in the in vitro group. Furthermore, the ratios of blood vessel volume density-to-placentome surface area were increased in the in vitro group. In conclusion, these findings are consistent with the concept that compensatory mechanisms exist in the vascular beds of placentas from bovine embryos produced in vitro.
本研究的目的是确定体外胚胎生产对妊娠后期牛胎盘血管生成和形态学的影响。体内产生的囊胚从超排的荷斯坦奶牛中回收。体外产生的囊胚是在体外成熟和受精的荷斯坦卵母细胞培养后获得的。将来自每个生产系统的单个囊胚移植到小母牛体内。在妊娠第222天回收胎儿和胎盘(体内组,n = 12;体外组,n = 12)。获取子叶和肉阜组织以定量血管内皮生长因子(VEGF)和过氧化物酶体增殖物激活受体γ(PPARγ)的mRNA和蛋白质。制备胎盘小叶的组织切片用于形态学分析。体外产生的胚胎的胎儿和胎盘比体内产生的胚胎更重。体外产生的胚胎的更多胎盘有过量的胎盘液。处理对子叶或肉阜组织中VEGF和PPARγ的mRNA表达没有影响。体外组和体内组之间,子叶和肉阜中VEGF蛋白的表达以及子叶中PPARγ蛋白的表达没有差异。然而,体外组肉阜中PPARγ蛋白的表达增加。与对照组相比,体外组子宫内膜的总表面积更大。相反,体外组胎盘小叶表面积百分比降低。体外产生的胚胎的胎盘小叶中胎儿绒毛和双核细胞体积密度降低。体外组母体肉阜中血管的比例组织体积增加。此外,体外组血管体积密度与胎盘小叶表面积的比率增加。总之,这些发现与以下概念一致,即体外产生的牛胚胎的胎盘血管床中存在补偿机制。