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Angiogenesis in day-30 bovine pregnancies derived from nuclear transfer

机译:来自核移植的第30天牛妊娠的血管生成

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Impaired placental angiogenesis during early pregnancy may result in placental defects that adversely affect development of nuclear-transfer (NT) embryos later in pregnancy. These experiments were designed to quantify and compare development of placental microvasculature and expression of genes associated with angiogenesis, including members of the VEGF and angiopoietin (Ang) families, in maternal and embryonic placental tissues of day 30 bovine concepti derived from NT or in vitro fertilization (IVF) followed by in vivo development to the blastocyst stage in the sheep oviduct. Microvascular volume density (MVD) within the caruncular tissues, as determined using Periodic Acid-Schiff's staining as well as immunohistochemical staining for von Willebrand's factor, was not different between NT- and IVF- derived pregnancies. Expression of genes implicated in angiogenic mechanisms, including VEGF-A and -C, placental growth factor (PlGF), VEGF receptors (Flt-1, Flk-1, and Flt-4), angiopoietin-1 (Ang1), Ang2, Tie1, Tie2, and hypoxia-inducible factor (HIF), were determined. In chorio-allantoic membranes, levels of PlGF transcripts were significantly lower in NT- than IVF-derived tissues (p < 0.05), whereas HIF-1alpha transcription in chorio-allantoic membranes of cloned concepti was higher at p < 0.10. Caruncular expression of HIF-1alpha and Ang1 also was increased in NT-derived pregnancies at p
机译:妊娠早期胎盘血管生成受损可能会导致胎盘缺陷,从而不利地影响妊娠后期核移植(NT)胚胎的发育。设计这些实验的目的是量化和比较第30天来自NT或体外受精的母胎和胚胎胎盘组织中胎盘微血管的发育以及与血管生成相关的基因的表达,包括VEGF和血管生成素(Ang)家族的成员(IVF),然后体内发育到绵羊输卵管的胚泡期。使用高碘酸希夫氏染色法和冯·威布兰德因子的免疫组化染色法测定的肾小管组织内的微血管体积密度(MVD)在NT和IVF妊娠之间没有差异。涉及血管生成机制的基因的表达,包括VEGF-A和-C,胎盘生长因子(PlGF),VEGF受体(Flt-1,Flk-1和Flt-4),血管生成素-1(Ang1,Ang2,Tie1) ,Tie2和缺氧诱导因子(HIF)被确定。在绒毛膜尿膜中,NT-组织中PlGF转录物水平显着低于IVF来源的组织(p <0.05),而克隆概念的绒毛膜尿酸膜中的HIF-1alpha转录水平更高,p <0.10。在NT衍生的妊娠中,HIF-1alpha和Ang1的圆盘状表达也增加了,p = 0.10。 VEGF-A和Flt-1受体对肾小管组织的免疫组织化学染色显示,NT和IVF孕妇的蛋白质表达差异很小。这些结果表明,在妊娠第30天,大多数血管生成因子的表达并未因NT程序而改变;然而,鉴于NT来源的牛胚胎中胎盘血管发育受损的报道,在胎盘早期发育和整个妊娠过程中,可能随后发生血管新生的紊乱。 HIF-1alpha基因的表达在克隆的概念的母体和绒毛膜尿道组织中升高可能表明NT衍生的概念的早期胎盘组织中存在普遍的低氧状况,这可能对胎盘的后续发育产生不利影响。

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