Romek M, Gajda B, Rolka M, Smorąg Z
Department of Cytology and Histology, Institute of Zoology, Jagiellonian University, Kraków, Poland.
Reprod Domest Anim. 2011 Jun;46(3):471-80. doi: 10.1111/j.1439-0531.2010.01691.x. Epub 2010 Oct 19.
Mitochondria are important determinants of developmental competence for oocytes and embryos owing to their central role in cellular metabolism, yet mitochondrial activity and morphometry during early porcine development have not been quantified. In this study, we examined the membrane potential Δψ(m) and the surface density Sv(in,m) of the inner mitochondrial membrane in pig oocytes and pre-implantation embryos using fluorescent probes and confocal microscopy. Mitochondria and their cristae were also examined by transmission electron microscope. Δψ(m) was consistently low from immature oocytes up to morulae and increased significantly in the early blastocyst before decreasing at the expanded blastocyst stage. This stage-dependent pattern of Δψ(m) changes differs from that reported for other mammals. We also determined that Δψ(m) is lower in cultured when compared to non-cultured porcine early blastocysts. Sv(in,m) was higher in immature oocytes than mature oocytes and remained constant up to the 4- to 8-cell embryo stage. It increased significantly at morula and early blastocyst stages. No differences in Sv(in,m) were found between developmentally matched non-cultured and cultured embryos. These results indicate that the inner mitochondrial membrane potential and surface density change significantly during pre-implantation porcine development in relation to metabolic alterations of the embryo. It is possible that modification of Δψ(m) by manipulating culture conditions may improve the performance of embryos that develop in vitro.
线粒体因其在细胞代谢中的核心作用,是卵母细胞和胚胎发育能力的重要决定因素,但猪早期发育过程中的线粒体活性和形态计量学尚未得到量化。在本研究中,我们使用荧光探针和共聚焦显微镜检查了猪卵母细胞和植入前胚胎中线粒体膜电位Δψ(m)和线粒体内膜的表面密度Sv(in,m)。还通过透射电子显微镜检查了线粒体及其嵴。从未成熟卵母细胞到桑椹胚,Δψ(m)一直较低,在早期囊胚中显著增加,然后在扩张囊胚阶段下降。这种依赖阶段的Δψ(m)变化模式与其他哺乳动物的报道不同。我们还确定,与未培养的猪早期囊胚相比,培养的囊胚中Δψ(m)较低。未成熟卵母细胞中的Sv(in,m)高于成熟卵母细胞,直到4至8细胞胚胎阶段保持恒定。在桑椹胚和早期囊胚阶段显著增加。在发育匹配的未培养和培养胚胎之间未发现Sv(in,m)有差异。这些结果表明,在猪植入前发育过程中,线粒体内膜电位和表面密度随胚胎代谢改变而显著变化。通过控制培养条件改变Δψ(m)可能会改善体外发育胚胎的性能。