Harvey M B, Arcellana-Panlilio M Y, Zhang X, Schultz G A, Watson A J
Department of Medical Biochemistry, University of Calgary, AB, Canada.
Biol Reprod. 1995 Sep;53(3):532-40. doi: 10.1095/biolreprod53.3.532.
Preimplantation embryos from a variety of mammalian species contrast markedly in their response to culture in vitro. Murine preimplantation embryos display a wider tolerance than other mammalian species to culture environments, and this has contributed to the development of several effective defined culture media. Embryo coculture on somatic cells remains the most effective method of supporting reasonable rates of bovine preimplantation development in vitro. The patterns of gene expression for several antioxidant enzymes during preimplantation murine and bovine development were examined by use of the reverse transcription-polymerase chain reaction technique to determine whether the differential developmental capacity of mammalian preimplantation embryos in culture may reflect variations in the patterns of expression for a series of antioxidant enzymes. Transcripts for catalase, CuZn-containing superoxide dismutase (CuZn-SOD), Mn-SOD, glutathione peroxidase (GPX), and glutamylcysteine synthetase (GCS) were detected in mouse embryos at all stages of development regardless of in vivo or in vitro development. Preimplantation cow embryos produced by in vitro procedures expressed mRNAs for catalase, CuZn-SOD and GPX, whereas transcripts for Mn-SOD were not detected at any stage. GCS transcripts, although present in stages up to the morula, were not detected in cow blastocysts. Analysis of antioxidant gene expression in both bovine primary oviductal cell monolayer cultures and nonattached, ciliated oviductal cell vesicle cultures revealed a constitutive pattern of expression of all five enzymes for the 8-day culture interval. These experiments suggest that differences in gene expression may contribute to the variation in the ability of embryos to develop in vitro with respect to levels of oxygen and dependence on coculture.
来自多种哺乳动物物种的植入前胚胎在体外培养反应方面存在显著差异。与其他哺乳动物物种相比,小鼠植入前胚胎对培养环境具有更广泛的耐受性,这促进了几种有效的限定培养基的开发。体细胞上的胚胎共培养仍然是支持牛体外植入前发育达到合理比率的最有效方法。利用逆转录-聚合酶链反应技术检测了小鼠和牛植入前发育过程中几种抗氧化酶的基因表达模式,以确定哺乳动物植入前胚胎在培养中发育能力的差异是否可能反映了一系列抗氧化酶表达模式的变化。无论体内还是体外发育,在小鼠胚胎发育的所有阶段都检测到了过氧化氢酶、含铜锌超氧化物歧化酶(CuZn-SOD)、锰超氧化物歧化酶(Mn-SOD)、谷胱甘肽过氧化物酶(GPX)和谷氨酰半胱氨酸合成酶(GCS)的转录本。体外培养产生的植入前牛胚胎表达了过氧化氢酶、CuZn-SOD和GPX的mRNA,而在任何阶段都未检测到Mn-SOD的转录本。GCS转录本虽然在桑椹胚之前的阶段存在,但在牛囊胚中未检测到。对牛原代输卵管细胞单层培养物和未附着的纤毛输卵管细胞囊泡培养物中的抗氧化基因表达分析表明,在8天的培养间隔内,所有五种酶的表达呈组成型模式。这些实验表明,基因表达的差异可能导致胚胎在体外发育能力在氧气水平和对共培养的依赖性方面的差异。