Department of Animal Science, Division of Applied Life Science (BK21), Graduate School of Gyeongsang National University, Jinju 660-701, Republic of Korea.
Anim Reprod Sci. 2013 Nov 1;142(1-2):19-27. doi: 10.1016/j.anireprosci.2013.08.012. Epub 2013 Sep 9.
The aim of this study was to investigate the developmental rate, lipid and mitochondrial distribution and gene expression in oocyte-derived embryos selected on the basis of brilliant cresyl blue (BCB) staining. Lipid content and mitochondrial distribution in Day 8 blastocysts were evaluated by fluorescence intensity, while gene expression was analyzed by real-time PCR. The proportion of blastocysts (30.9%) was greater (P<0.05) in BCB+ than in BCB- oocytes (13%) but not different (P>0.05) from the control group (28.2%). Total cell number was also greater in BCB+ (155.1 ± 36.2) than in BCB- (116.6 ± 40.5) and control (127.5 ± 35.7) blastocysts. Furthermore, the apoptotic cell number was less in BCB+ (3.7 ± 4.4) than in BCB- blastocysts (8.7 ± 8.7) but not different from the control group (5.9 ± 3.9). BCB+ embryos contained more mitochondria compared to BCB- embryos (P<0.05). There was no significant difference in intercellular lipid accumulation in embryos from all groups. Interferon-τ (IFNτ), transforming growth factor β1 (TGFB1) and secreted seminal-vesicle Ly-6 protein 1 (SSLP1) gene expression was greater in BCB+ than in BCB- blastocysts. By contrast, Bcl2-associated X protein (BAX) and heterogeneous nuclear ribonucleoprotein A2/B1 (HNRNPA2B1) gene expression was greater in BCB- than in BCB+ and control embryos. In conclusion, oocyte-derived embryos selected on the basis of BCB staining showed differences in developmental rate, quality, mitochondrial content and target gene expression compared to control embryos.
本研究旨在探讨根据亮蓝（BCB）染色选择的卵母细胞衍生胚胎的发育速度、脂质和线粒体分布以及基因表达。通过荧光强度评估第 8 天囊胚的脂质含量和线粒体分布，同时通过实时 PCR 分析基因表达。BCB+卵母细胞的囊胚比例（30.9%）高于 BCB-卵母细胞（13%）（P<0.05），但与对照组（28.2%）无差异（P>0.05）。BCB+囊胚的总细胞数也多于 BCB-（116.6±40.5）和对照组（127.5±35.7）囊胚（P<0.05）。此外，BCB+囊胚的凋亡细胞数（3.7±4.4）少于 BCB-囊胚（8.7±8.7），但与对照组无差异（5.9±3.9）。与 BCB-胚胎相比，BCB+胚胎含有更多的线粒体（P<0.05）。三组胚胎间细胞内脂质积累无显著差异。BCB+胚胎的干扰素-τ（IFNτ）、转化生长因子β1（TGFB1）和分泌性精囊 Ly-6 蛋白 1（SSLP1）基因表达高于 BCB-胚胎。相反，BCB-胚胎的 Bcl2 相关 X 蛋白（BAX）和异质性核核糖核蛋白 A2/B1（HNRNPA2B1）基因表达高于 BCB+和对照组胚胎。综上所述，与对照组胚胎相比，根据 BCB 染色选择的卵母细胞衍生胚胎在发育速度、质量、线粒体含量和靶基因表达方面存在差异。
