Guillomot M, Campion E, Prézelin A, Sandra O, Hue I, Le Bourhis D, Richard C, Biase F H, Rabel C, Wallace R, Lewin H, Renard J-P, Jammes H
INRA, UMR1198 Biologie du Développement et Reproduction, F-78350 Jouy-en-Josas, France.
INRA, UMR1198 Biologie du Développement et Reproduction, F-78350 Jouy-en-Josas, France.
Placenta. 2014 Sep;35(9):737-47. doi: 10.1016/j.placenta.2014.06.366. Epub 2014 Jun 26.
Alteration of expression of various genes including extracellular matrix components, have been suggested to play major role in the placental pathologies after somatic cloning in mammals. The objectives of the present study were to analyze pattern of expression (mRNA and protein) of the small leucine-rich proteoglycan, Decorin in association with Type I Collagen and Fibronectin in bovine placental tissues from normal and clone pregnancies.
Genotyping and allelic expression of Decorin were determined by Sanger sequencing. The expression patterns of Decorin, Type I collagen and Fibronectin 1 were analyzed by quantitative RT-qPCR and combined in situ hybydization (ISH) and immunohistochemistry (IHC) in endometrial and placental tissues from D18 to term from artificially inseminated and somatic cloning pregnancies.
The expression levels of DCN increased in the AI endometrial stroma and chorionic mesenchyme during implantation and declined during placentome growth until term. Combined ISH and IHC revealed an unexpected discrepancy mRNA and protein tissue distribution. Moreover, Decorin was maintained in the placentome tissues from SCNT pregnancies while both mRNA and protein were absent in AI derived placenta.
In bovine, the pattern of expression of Decorin exhibits significant changes during placental formation. Downregulation of Decorin is associated with proliferation, remodeling and vascularization of placental tissues. These observations reinforces the putative role of Decorin in these processes.
These observations suggest that Decorin is involved in placental growth and that dysregulation of its expression is associated with placental abnormalities in SCNT derived pregnancy.
包括细胞外基质成分在内的各种基因表达的改变,被认为在哺乳动物体细胞克隆后的胎盘病理过程中起主要作用。本研究的目的是分析正常妊娠和克隆妊娠的牛胎盘组织中,富含亮氨酸的小分子蛋白聚糖核心蛋白聚糖(Decorin)与I型胶原蛋白和纤连蛋白的表达模式(mRNA和蛋白质)。
通过桑格测序确定核心蛋白聚糖的基因分型和等位基因表达。采用定量逆转录-定量聚合酶链反应(RT-qPCR)分析核心蛋白聚糖、I型胶原蛋白和纤连蛋白1的表达模式,并结合原位杂交(ISH)和免疫组织化学(IHC),对人工授精和体细胞克隆妊娠从妊娠18天到足月的子宫内膜和胎盘组织进行分析。
在人工授精妊娠中,植入期间子宫内膜基质和绒毛膜间充质中核心蛋白聚糖(DCN)的表达水平升高,在胎盘小叶生长至足月期间下降。ISH和IHC联合检测显示,mRNA和蛋白质的组织分布存在意外差异。此外,体细胞核移植(SCNT)妊娠的胎盘小叶组织中维持有核心蛋白聚糖,而人工授精来源的胎盘中mRNA和蛋白质均不存在。
在牛中,核心蛋白聚糖的表达模式在胎盘形成过程中表现出显著变化。核心蛋白聚糖的下调与胎盘组织的增殖、重塑和血管生成有关。这些观察结果强化了核心蛋白聚糖在这些过程中的假定作用。
这些观察结果表明,核心蛋白聚糖参与胎盘生长,其表达失调与体细胞核移植妊娠中的胎盘异常有关。