Lazzari Giovanna, Wrenzycki Christine, Herrmann Doris, Duchi Roberto, Kruip Theo, Niemann Heiner, Galli Cesare
Laboratorio di Tecnologie della Riproduzione, Cremona, Italy.
Biol Reprod. 2002 Sep;67(3):767-75. doi: 10.1095/biolreprod.102.004481.
The large offspring syndrome (LOS) is observed in bovine and ovine offspring following transfer of in vitro-produced (IVP) or cloned embryos and is characterized by a multitude of pathologic changes, of which extended gestation length and increased birthweight are predominant features. In the present study, we used bovine blastocysts to analyze cellular parameters, i.e., the number of cells in Day 7 blastocysts and the size of Day 12 elongating blastocysts, and molecular parameters, i.e., the relative abundance of developmentally important genes: glucose transporter (Glut) 1, Glut-2, Glut-3, Glut-4, heat shock protein (Hsp) 70.1, Cu/Zn-superoxide dismutase (SOD), histone H4.1, basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF) I receptor (R), and IGFII-R. Some blastocysts were produced by in vitro maturation and fertilization followed by in vitro culture in synthetic oviduct fluid medium supplemented with BSA or human serum or by in vivo culture in the sheep oviduct. Other blastocysts were derived in vivo from the uterine horns of superovulated donors. The findings made in the early embryos were related to a representative number of calves obtained from each production system and from artificial insemination (AI). In vitro culture of bovine embryos in the presence of high concentrations of serum or BSA significantly increased the number of cells in Day 7 blastocysts, the size of blastocysts on Day 12, and the relative abundance of the transcripts for Hsp70.1, Cu/Zn-SOD, Glut-3, Glut-4, bFGF, and IGFI-R when compared with embryos from the in vivo production groups. Birthweights of calves derived from IVP embryos were significantly higher than those of calves derived from sheep oviduct culture, superovulation, or AI. The results support the hypothesis that persistence of early deviations in development is causally involved in the incidence of LOS, in particular in increased birthweights. The cellular and molecular parameters analyzed in this study can be considered early markers of LOS in cattle.
在体外生产（IVP）或克隆胚胎移植后的牛和羊后代中观察到巨大后代综合征（LOS），其特征是有多种病理变化，其中妊娠期延长和出生体重增加是主要特征。在本研究中，我们使用牛囊胚来分析细胞参数，即第7天囊胚中的细胞数量和第12天伸长囊胚的大小，以及分子参数，即发育重要基因的相对丰度：葡萄糖转运蛋白（Glut）1、Glut - 2、Glut - 3、Glut - 4、热休克蛋白（Hsp）70.1、铜/锌超氧化物歧化酶（SOD）、组蛋白H4.1、碱性成纤维细胞生长因子（bFGF）、胰岛素样生长因子（IGF）I受体（R）和IGFII - R。一些囊胚是通过体外成熟和受精，然后在补充有牛血清白蛋白或人血清的合成输卵管液培养基中进行体外培养产生的，或者是在绵羊输卵管中进行体内培养产生的。其他囊胚是从超排供体的子宫角体内获得的。在早期胚胎中得到的结果与从每个生产系统以及人工授精（AI）获得的代表性数量的犊牛有关。与体内生产组的胚胎相比，在高浓度血清或牛血清白蛋白存在下对牛胚胎进行体外培养显著增加了第7天囊胚中的细胞数量、第12天囊胚的大小以及Hsp70.1、铜/锌 - SOD、Glut - 3、Glut - 4、bFGF和IGFI - R转录本的相对丰度。来自IVP胚胎的犊牛出生体重显著高于来自绵羊输卵管培养、超排或人工授精的犊牛。结果支持这样的假设，即发育早期偏差的持续存在与LOS的发生率有因果关系，特别是与出生体重增加有关。本研究中分析的细胞和分子参数可被视为牛LOS的早期标志物。
