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在人类和小鼠卵母细胞成熟的最后阶段,编码抗氧化酶的基因表达情况。

Expression of genes encoding antioxidant enzymes in human and mouse oocytes during the final stages of maturation.

作者信息

El Mouatassim S, Guérin P, Ménézo Y

机构信息

Laboratoire Marcel Mérieux, Cytogénétique, Avenue Tony Garnier BP 7322, 69357 Lyon, France.

出版信息

Mol Hum Reprod. 1999 Aug;5(8):720-5. doi: 10.1093/molehr/5.8.720.

Abstract

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 MII 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.

摘要

研究了参与自由基防御的五种酶(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))在人和小鼠卵母细胞中的mRNA表达。在小鼠中,收集处于不同成熟阶段的卵母细胞,以确定这些转录本的储存情况。对于人类,分析了在卵胞浆内单精子注射(ICSI)过程中收集的生发泡(GV)卵母细胞以及受精失败的中期II(MII)卵母细胞。比较人和小鼠,以确定小鼠和人类植入前胚胎在培养中的发育能力差异是否可以由这些酶表达模式的变化来解释。使用逆转录聚合酶链反应(RT-PCR)检测这些酶的mRNA表达。在小鼠中,无论成熟阶段如何,均可检测到所有转录本(过氧化氢酶除外)。GV和MII卵母细胞之间未检测到定性差异。在人类中,所有酶(过氧化氢酶除外)均在MII卵母细胞中表达,且Cu-Zn-SOD表达尤为高。GV期未检测到与GPX和Mn-SOD对应的转录本,仅在MII期检测到,这表明在GV和MII期之间可能发生储存。然而,在逆转录反应中使用针对GPX和Mn-SOD的3'端特异性引物而非寡聚(dT)(12-18)引物,已在GV期的人类卵母细胞中检测到这些抗氧化酶的转录本。这表明这些转录本存在成熟特异性多聚腺苷酸化。这些酶可被视为细胞质成熟的标志物。

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