Expression of Apoptosis Regulatory Genes and Incidence of Apoptosis in Different Morphological Quality Groups of In Vitro-produced Bovine Pre-implantation Embryos
Expression of Apoptosis Regulatory Genes and Incidence of Apoptosis in Different Morphological Quality Groups of In Vitro-produced Bovine Pre-implantation Embryos
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DOI：
10.1111/j.1439-0531.2009.01463.x
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发表时间：
2010-10-01
影响因子：
1.7
通讯作者：
Tesfaye, D.
中科院分区：
文献类型：
--
作者：
Melka, M. G.;Rings, F.;Tesfaye, D.
细胞凋亡发生在体内和体外产生的胚胎的早期发育过程中，被认为是胚胎丢失的原因之一。本研究的目的是，因此，调查阶段特异性表达谱的凋亡调控基因在三个质量组的体外生产的牛植入前胚胎，并分析细胞数量和DNA片段与这些基因的表达之间的关系。分析9种促凋亡基因（Bax、caspase-9、Bcl-xs、P53、Caspase-3和Fas）和抗凋亡基因（Bcl-w和Mcl-1）mRNA的相对丰度。分别采用细胞差异染色和末端脱氧核苷酸转移酶介导的dUTP-生物素缺口末端标记法分析细胞数量的变化和检测凋亡细胞核。植入前胚胎质量差的Bax和Caspase-3基因表达显著高于相同发育阶段形态学良好的胚胎（p < 0.05）。质量好的未成熟卵母细胞中Mcl-1的表达显著高于质量差的未成熟卵母细胞。此外，较高的DNA片段被证明在形态质量差的囊胚。总之，我们的研究表明，Bax，caspase-3和Mcl-1可以作为胚胎质量的潜在标志物，以评估在体外生产的牛胚胎。需要进一步的研究来调查可用于评估体内衍生胚胎的特定分子特征。
Apoptosis occurs during early development in both in vivo- and in vitro-produced embryos, and is considered as one of the causes of embryonic loss. The objectives of this study were, therefore, investigating stage-specific expression profiles of apoptosis regulatory genes in three quality groups of in vitro-produced bovine pre-implantation embryos; and analysing the relationship between cell number and DNA fragmentation with expressions of those genes. The relative abundance of mRNA of 9 pro- (Bax, caspase-9, Bcl-xs, P53, Caspase-3 and Fas) and anti- (Bcl-w and Mcl-1) apoptotic genes was analysed. Differential cell staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling were performed to analyse the variation in cell numbers and detect apoptotic nuclei respectively. Expression of Bax and Caspase-3 genes was significantly (p < 0.05) higher in poor quality preimplantation embryos as compared with that of morphologically good quality embryos of the same developmental stages. Moreover, Mcl-1 expression was significantly higher in good quality immature oocytes than that in the poor quality group. Moreover, higher DNA fragmentation was evidenced in morphologically poor quality blastocysts. In conclusion, our study demonstrates that Bax, caspase-3 and Mcl-1 can be used as potential markers of embryo quality to evaluate in vitro-produced bovine embryos. Further studies are required to investigate specific molecular signatures that can be used in evaluating in vivo-derived embryos.