Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning, 530001, China; State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Animal Science and Technology, Guangxi University, Nanning, Guangxi, 530004, China.
Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning, 530001, China.
Theriogenology. 2021 Jul 15;169:14-20. doi: 10.1016/j.theriogenology.2021.03.025. Epub 2021 Apr 13.
This study was performed to investigate the difference in developmental competence of oocytes derived from ovum pick-up (OPU) and slaughterhouse ovaries (SLH), and its underlying mechanisms. The OPU and SLH oocytes were in-vitro maturated and fertilized to produce blastocysts, and these blastoycsts were collected to explore the expression of key genes for developmental potential and telomere (Oct-4, Sox2, Nanog, Cdx2, Gata3, E-cadherin, β-catenin, TERT, TERF1 and TERF2). The results showed that both the cleavage and blastocyst rates were significantly higher for the OPU group (68.31%, 39.48%, respectively) than SLH group (57.59%, 26.50%, respectively) (P < 0.01). The relative mRNA abundances of Sox2, Oct-4, Nanog and E-cadherin were significantly higher in the OPU blastocysts than the SLH ones (P < 0.01). Protein expression analysis by Western blot and immunofluorescence also revealed that the expression of E-cadherin and Sox2 was significantly higher in OPU blastocysts than SLH ones. However, there was no significant differences between the two groups in the expression of Cdx2, β-catenin, Gata3, TERT, TERF1, TERF2. These results imply oocyte sources modify the expression of development and adhesion related genes in blastocysts, which may elucidate a possible reasoning for the low development competence of buffalo SLH embryos.
本研究旨在探讨卵母细胞抽吸(OPU)和屠宰场卵巢(SLH)来源的卵母细胞在发育能力上的差异及其潜在机制。将 OPU 和 SLH 卵母细胞进行体外成熟和受精,以产生囊胚,并收集这些囊胚,以探讨发育潜能和端粒的关键基因的表达(Oct-4、Sox2、Nanog、Cdx2、Gata3、E-cadherin、β-catenin、TERT、TERF1 和 TERF2)。结果表明,OPU 组的卵裂率和囊胚率均显著高于 SLH 组(分别为 68.31%和 39.48%)(P<0.01)。OPU 囊胚中 Sox2、Oct-4、Nanog 和 E-cadherin 的相对 mRNA 丰度明显高于 SLH 囊胚(P<0.01)。Western blot 和免疫荧光蛋白表达分析也表明,E-cadherin 和 Sox2 的表达在 OPU 囊胚中明显高于 SLH 囊胚。然而,两组在 Cdx2、β-catenin、Gata3、TERT、TERF1、TERF2 的表达上无显著差异。这些结果表明,卵母细胞来源改变了囊胚中发育和黏附相关基因的表达,这可能解释了水牛 SLH 胚胎发育能力低的原因。