Mamo S, Ponsuksili S, Wimmers K, Gilles M, Schellander K
Institute of Animal Breeding Sciences, University of Bonn, Bonn, Germany.
Reprod Domest Anim. 2005 Apr;40(2):177-83. doi: 10.1111/j.1439-0531.2005.00579.x.
Retinoid X receptors are frequently implicated in their role in embryonic development. However, there is no report regarding their expression in embryos with different qualities. In this study, expression pattern and levels of retinoid X receptor transcripts in different qualities and stages of in vitro-produced pre-implantation bovine embryos were examined using real-time polymerase chain reaction (PCR) techniques. Four independent cDNA pools per sample were prepared from mRNAs isolated from different developmental-stage embryos, after classifying them to be of good and bad quality. Primers were also designed to amplify products with PCR. Based on analysis, except retinoid X receptor gamma (RXRG) transcripts, the remaining two types (alpha and beta forms) were expressed in all samples with significantly higher (p < or = 0.05) levels until the eight-cell stage compared with the stages thereafter. However, the transcript levels for bad-quality embryos were consistently lower in all cases in comparison with good-quality embryos. RXRG was identified in 39% of the samples, but all in the pre-embryonic genome activation development stage. Therefore, strong expression patterns of these transcripts in earlier stages indicates their possible role during the maternal phase of embryo development, and the variations of copy number in embryos with different qualities substantiates their possible candidacy as potential quality markers. Moreover, identifying the transcript variations in different qualities and expression of RXRG at these stages is a novel input to the pre-implantation-stage knowledge.
维甲酸X受体在胚胎发育中的作用经常被提及。然而,关于它们在不同质量胚胎中的表达尚无报道。在本研究中,使用实时聚合酶链反应(PCR)技术检测了体外生产的植入前牛胚胎在不同质量和阶段的维甲酸X受体转录本的表达模式和水平。在将不同发育阶段的胚胎分为优质和劣质后,从分离的mRNA中为每个样本制备了四个独立的cDNA池。还设计了引物用于通过PCR扩增产物。基于分析,除了维甲酸X受体γ(RXRG)转录本外,其余两种类型(α和β形式)在所有样本中均有表达,直到八细胞期,其水平与之后的阶段相比显著更高(p≤0.05)。然而,与优质胚胎相比,劣质胚胎的转录水平在所有情况下始终较低。在39%的样本中鉴定出了RXRG,但均处于胚胎前基因组激活发育阶段。因此,这些转录本在早期阶段的强表达模式表明它们在胚胎发育的母源阶段可能发挥的作用,不同质量胚胎中拷贝数的变化证实了它们作为潜在质量标记的可能候选资格。此外,确定不同质量下的转录本变化以及这些阶段RXRG的表达是对植入前阶段知识的新补充。