National Foundation for Fertility Research, Lone Tree, CO 80124, USA.
J Anim Sci. 2011 Nov;89(11):3561-71. doi: 10.2527/jas.2011-4193. Epub 2011 Jun 16.
The developmental competence of oocytes is progressively attained as females approach puberty. The poor quality of prepubertally derived oocytes suggests that essential processes during cytoplasmic maturation have not been completed. The objective of this experiment was to identify genes in oocytes that are associated with good (cyclic females) and poor (prepubertal females) developmental competence. Development to the blastocyst stage in vitro was significantly decreased in oocytes derived from prepubertal females compared with cyclic females (5.26 and 12.86%, respectively). Approximately 10% of the oocyte transcriptome was differentially expressed between in vitro-matured oocytes derived from cyclic and prepubertal females (P < 0.05); 58% of differentially expressed genes had increased transcript abundance in oocytes derived from cyclic females. Genes involved in the metabolism and regulation of biological processes had increased transcript abundance in oocytes derived from cyclic females, whereas genes involved in translation were increased in prepubertally derived oocytes. Quantitative PCR confirmed differential expression (P < 0.05) for 6 out of 11 selected genes [DPYD (dihydropyrimidine dehydrogenase), RDH11 (retinol dehydrogenase 11), SFRS4 (serine/arginine-rich splicing factor 4), SFRS7 (serine/arginine-rich splicing factor 7), TL4 (transcribed loci 4), and TOP2B (topoisomerase II β)] that were differentially expressed with greater than a 2-fold change by microarray, although 3 of these genes, DPYD, TL4, and TOP2B, were in opposing directions by the 2 methods. In conclusion, expression of multiple genes involved in metabolism and translation was significantly altered in oocytes from prepubertal females compared with cyclic females, which was associated with reduced in vitro development to the blastocyst stage. These genes may represent important cellular mechanisms that regulate oocyte quality.
卵母细胞的发育能力随着女性接近青春期而逐渐获得。未成熟来源的卵母细胞质量较差,表明细胞质成熟过程中的基本过程尚未完成。本实验的目的是鉴定与良好(循环女性)和不良(青春期前女性)发育能力相关的卵母细胞中的基因。与循环女性相比,来自青春期前女性的卵母细胞体外发育至囊胚阶段的能力显著降低(分别为 5.26%和 12.86%)。在体外成熟的卵母细胞中,来自循环和青春期前女性的卵母细胞之间有大约 10%的卵母细胞转录组存在差异表达(P<0.05);在循环女性来源的卵母细胞中,58%差异表达的基因转录丰度增加。参与代谢和生物过程调节的基因在循环女性来源的卵母细胞中转录丰度增加,而参与翻译的基因在青春期前来源的卵母细胞中增加。定量 PCR 证实了 11 个选定基因中的 6 个(DPYD(二氢嘧啶脱氢酶)、RDH11(视黄醇脱氢酶 11)、SFRS4(丝氨酸/精氨酸丰富剪接因子 4)、SFRS7(丝氨酸/精氨酸丰富剪接因子 7)、TL4(转录基因 4)和 TOP2B(拓扑异构酶 IIβ))的差异表达(P<0.05),这些基因在微阵列中差异表达超过 2 倍,但这 3 个基因,DPYD、TL4 和 TOP2B,两种方法的方向相反。总之,与循环女性相比,青春期前女性的卵母细胞中涉及代谢和翻译的多个基因的表达明显改变,这与体外发育至囊胚阶段的能力降低有关。这些基因可能代表调节卵母细胞质量的重要细胞机制。