Gjørret Jakob O, Knijn Hiemke M, Dieleman Steph J, Avery Birthe, Larsson Lars-Inge, Maddox-Hyttel Poul
Department of Anatomy and Physiology, Royal Veterinary and Agricultural University, 1870 Frederiksberg C, Denmark.
Biol Reprod. 2003 Oct;69(4):1193-200. doi: 10.1095/biolreprod.102.013243. Epub 2003 May 28.
The postimplantation developmental potential of embryos can be affected by various forms of cell death, such as apoptosis, at preimplantation stages. However, correct assessment of apoptosis is needed for adequate inference of the developmental significance of this process. This study is the first to investigate the independent chronological occurrence of apoptotic changes in nuclear morphology and DNA degradation (detected by the TUNEL reaction) and incidences of nuclei displaying these features at various preimplantation stages of bovine embryos produced both in vivo and in vitro. Different elements of apoptosis were observed at various developmental stages and appeared to be differentially affected by in vitro production. Nuclear condensation was observed from the 6-cell stage in vitro and the 8-cell stage in vivo, whereas the TUNEL reaction was first observed at the 6-cell stage in vitro and the 21-cell stage in vivo. Morphological signs of other forms of cell death were also observed in normally developing embryos produced both in vivo and in vitro. The onset of apoptosis seems to be developmentally regulated in a stage-specific manner, but discrete features of the apoptotic process may be differentially regulated and independently modulated by the mode of embryo production. Significant differences in indices of various apoptotic features were not evident between in vivo- and in vitro-produced embryos at the morula stage, but such differences could be observed at the blastocyst stage, where in vitro production was associated with a higher degree of apoptosis in the inner cell mass.
胚胎植入后的发育潜能可能会受到植入前阶段各种形式的细胞死亡(如凋亡)的影响。然而,为了充分推断这一过程的发育意义,需要对凋亡进行正确评估。本研究首次调查了核形态和DNA降解(通过TUNEL反应检测)中凋亡变化的独立时间顺序发生情况,以及在体内和体外产生的牛胚胎的各个植入前阶段显示这些特征的细胞核发生率。在不同发育阶段观察到凋亡的不同要素,并且似乎受到体外生产的不同影响。在体外6细胞阶段和体内8细胞阶段观察到核浓缩,而TUNEL反应在体外6细胞阶段和体内21细胞阶段首次观察到。在体内和体外产生的正常发育胚胎中也观察到其他形式细胞死亡的形态学迹象。凋亡的起始似乎以阶段特异性方式受到发育调控,但凋亡过程的离散特征可能受到胚胎生产方式的不同调控和独立调节。在桑椹胚阶段,体内和体外产生的胚胎之间各种凋亡特征指标的显著差异不明显,但在囊胚阶段可以观察到这种差异,体外生产与内细胞团中更高程度的凋亡相关。