Expression of genes encoding antioxidant enzymes in human and mouse oocytes during the final stages of maturation.
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.
El Mouatassim
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Laboratoire Marcel Mérieux, Cytogénétique, Avenue Tony Garnier BP 7322, 69357 Lyon, France.
Guérin
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Ménézo
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D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D001755:Blastocyst / Q000201:enzymology; D002374:Catalase / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005609:Free Radicals / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D005721:Glutamate-Cysteine Ligase / Q000235:genetics; D005979:Glutathione Peroxidase / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D009865:Oocytes / Q000201:enzymology* / Q000254:growth & development* / Q000378:metabolism; D009866:Oogenesis / Q000235:genetics / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics
D000975:Antioxidants; D017931:DNA Primers; D005609:Free Radicals; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D005721:Glutamate-Cysteine Ligase