Expression of genes encoding antioxidant enzymes in human and mouse oocytes during the final stages of maturation
Expression of genes encoding antioxidant enzymes in human and mouse oocytes during the final stages of maturation
复制标题
DOI：
10.1093/molehr/5.8.720
复制
发表时间：
1999-08-01
影响因子：
4
通讯作者：
Ménézo, Y
中科院分区：
文献类型：
--
作者：
El Mouatassim, S;Guérin, P;Ménézo, Y
在人类和小鼠卵母细胞中研究了五种酶的 mRNA 表达：过氧化氢酶、铜锌超氧化物歧化酶 (Cu-Zn-SOD)、锰超氧化物歧化酶 (Mn-SOD)、谷胱甘肽过氧化物酶 (GPX) 和 γ-谷氨酰半胱氨酸合成酶 (GCS)，每种酶都参与对抗自由基的保护。在小鼠中，收集不同成熟阶段的卵母细胞以确定这些转录物的储存。对于人类，分析了胞浆内单精子注射 (ICSI) 过程中收获的生发囊 (GV) 卵母细胞和失败的受精中期 II (MII) 卵母细胞。为了确定培养物中小鼠和人类植入前胚胎的发育能力差异是否可以通过这些酶的表达模式的变化来解释，对人和小鼠进行了比较，使用逆转录聚合酶链式反应（RT-PCR）检查了这些酶的 mRNA 表达。在小鼠中，无论成熟阶段如何，所有转录物（过氧化氢酶除外）均被检测到。 GV 和 MII 卵母细胞之间没有检测到质量差异。在人类中，所有酶（除过氧化氢酶外）均在 MII 卵母细胞中表达，其中 Cu-Zn-SOD 的表达尤其高。与 GPX 和 Mn-SOD 相对应的转录本在 GV 阶段未检测到，而仅在 Mil 阶段检测到，表明储存可能发生在 GV 和 MII 阶段之间。然而，在逆转录反应中使用 GPX 和 Mn-SOD 的 3' 末端特异性引物，而不是oligo(dT)(12-18) 引物，已在 GV 阶段的人类卵母细胞中检测到了这些抗氧化剂酶的转录本。这表明这些转录物存在成熟特异性多腺苷酸化。这些酶可以被认为是细胞质成熟的标志物。
The mRNA expression of five enzymes: catalase, Cu-Zn-superoxide dismutase (Cu-Zn-SOD), Mn-superoxide dismutase (Mn-SOD), glutathione peroxidase (GPX), and gamma-glutamylcysteine synthetase (GCS) each involved in protection against free radicals was studied in human and mouse oocytes. In the mouse, oocytes were collected at different stages of maturation in order to determine the storage of these transcripts. For the human, germinal vesicle (GV) oocytes harvested during intracytoplasmic sperm injection (ICSI) procedures and failed fertilized metaphase II (MII) oocytes were analysed. Human and mouse were compared in order to determine whether the differential developmental capacity of mouse and human preimplantation embryos in culture could be explained by the variations in the patterns of expression for these enzymes, mRNA expression for these enzymes was examined using reverse transcription-polymerase chain reaction (RT-PCR). In the mouse, all transcripts (except for catalase) were detected, whatever the maturation stage. No qualitative differences were detected between GV and MII oocytes. In human, all the enzymes (except for catalase) were expressed in MII oocytes and Cu-Zn-SOD was particularly highly expressed. Transcripts corresponding to GPX and Mn-SOD were not detected at GV stage but only at Mil stage, suggesting that storage could occur between GV and MII stages. However, using 3' end-specific primers for GPX and Mn-SOD, instead of the oligo(dT)(12-18) primer, for the reverse transcription reaction, the transcripts for these antioxidants enzymes have been detected in human oocytes at the GV stage. This suggests the presence of maturation-specific polyadenylation of these transcripts. These enzymes can be considered as markers of cytoplasmic maturation.