Mohammadi-Sangcheshmeh Abdollah, Veshkini Arash, Hajarizadeh Athena, Jamshidi-Adegani Fatemeh, Zhandi Mahdi, Abazari-Kia Amir Hossein, Cinar Mehmet Ulas, Soleimani Masoud, Gastal Eduardo L
Department of Animal and Poultry Science, College of Aburaihan, University of Tehran, Pakdasht, Tehran, Iran,
J Assist Reprod Genet. 2014 Aug;31(8):1089-98. doi: 10.1007/s10815-014-0264-6. Epub 2014 Jun 10.
To evaluate associations of glucose-6-phosphate dehydrogenase (G6PDH) activity in sheep oocytes with cytoplasmic lipid content, maturational competence, developmental competence to the blastocyst stage, and gene expression of certain molecular markers.
Before brilliant cresyl blue (BCB) staining test, oocytes were classified as high, middle, and low cytoplasmic lipid content (HCLC, MCLC, and LCLC) and after the test as having low or high G6PDH-activity (BCB(+) and BCB(-), respectively). After maturation in vitro, a group of oocytes were subjected to IVF followed by in vitro embryo culture and another group was used for evaluation of expression of candidate genes.
The cleavage and blastosyst rates were lowest (P < 0.05) in LCLC group, intermediate (P < 0.05) in MCLC group and highest (P < 0.05) in HCLC group. More (P < 0.05) oocytes in HCLC group were BCB(+), and higher (P < 0.05) maturation, cleavage, and blastocyst rates were seen for BCB(+) oocytes than the BCB(-) oocytes. Our gene expression data indicated that mRNA transcript abundance of ITGB2, pZP3, BMP15, and GDF9 genes was similar between BCB oocytes groups. However, the expression of ATP1A1 was higher (P < 0.05) for BCB(+) oocytes compared to BCB(-) oocytes. In addition, BAX transcript abundance was similar (P > 0.05) among BCB(+), BCB(-), and control groups, before and after maturation in vitro.
Activity of G6PDH in sheep oocytes is highly associated with lipid content, and compared with the morphological parameters might be a more precise and objective predictor for subsequent developmental competence in vitro.
评估绵羊卵母细胞中葡萄糖-6-磷酸脱氢酶（G6PDH）活性与细胞质脂质含量、成熟能力、发育至囊胚阶段的能力以及某些分子标志物的基因表达之间的关联。
在亮甲酚蓝（BCB）染色试验前，将卵母细胞分为高、中、低细胞质脂质含量组（分别为HCLC、MCLC和LCLC），试验后分为G6PDH活性低或高的组（分别为BCB(+)和BCB(-)）。体外成熟后，一组卵母细胞进行体外受精，随后进行体外胚胎培养，另一组用于评估候选基因的表达。
LCLC组的卵裂率和囊胚率最低（P < 0.05），MCLC组居中（P < 0.05），HCLC组最高（P < 0.05）。HCLC组中更多（P < 0.05）的卵母细胞为BCB(+)，BCB(+)卵母细胞的成熟率、卵裂率和囊胚率高于（P < 0.05）BCB(-)卵母细胞。我们的基因表达数据表明，ITGB2、pZP3、BMP15和GDF9基因的mRNA转录本丰度在BCB卵母细胞组之间相似。然而，与BCB(-)卵母细胞相比，BCB(+)卵母细胞中ATP1A1的表达更高（P < 0.05）。此外，体外成熟前后，BCB(+)、BCB(-)和对照组之间BAX转录本丰度相似（P > 0.05）。
绵羊卵母细胞中G6PDH的活性与脂质含量高度相关，与形态学参数相比，可能是后续体外发育能力更精确和客观的预测指标。
