Quantitative expression of candidate genes for developmental competence in bovine two-cell embryos.
Only competent oocytes are able to undergo complete maturation and normal embryonic development. Therefore, the identification of genes that are differentially expressed in competent oocytes would contribute to our understanding of the factors controlling competency. It is well known that time of cleavage after insemination in vitro is highly correlated with embryonic developmental potential and this can be used to distinguish between oocytes of different quality. The main objective of this study was to identify genes associated with competency and rapid cleavage. We examined the expression of 16 candidate genes (IDH, YEAF Cathepsin B, RAD50, TCP1 NCOR1, HUEL, STK6, ZNF403, AOP2, EEF1A1, Hsp90, Hsp40, AKR1B1, PGRMC1, and DMRT2) in early and late cleaving embryos, by real time PCR. These transcripts were derived from previous study in our laboratory using cDNA coming from a suppressive subtraction hybridization (SSH) between early cleaving versus late cleaving embryos spotted on a microarray slide. Of the 16 genes evaluated, 3 (IDH, YEAF, and H2A) showed statistical difference (P < 0.05) between early and late cleaving embryos. However, some genes such as Cathepsin B (P = 0.0677), RAD50 (P = 0.0899), and TCP1 (P = 0.0824) tended to show higher expression in the early cleaving than in the late cleaving embryo. In conclusion, we have identified three genes (YEAF, IDH, H2A) that were differentially expressed in the early cleaving embryos, and their expression can be associated with greater developmental competence.
Dode
Margot Alves Nunes
MA
Embrapa Recursos Genéticos e Biotecnologia, Brasília, DF, Brazil.
Dufort
Isabelle
I
Massicotte
Lyne
L
Sirard
Marc-André
MA
D000449:Aldehyde Reductase / Q000235:genetics; D000818:Animals; D002401:Cathepsin B / Q000235:genetics; D002417:Cattle; D002478:Cells, Cultured; D046508:Culture Techniques; D004268:DNA-Binding Proteins / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D047108:Embryonic Development / Q000235:genetics*; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics; D007521:Isocitrate Dehydrogenase / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D056971:Nuclear Receptor Co-Repressor 1; D015513:Oncogene Proteins / Q000235:genetics; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D010544:Peroxidases / Q000235:genetics; D054465:Peroxiredoxin VI; D054464:Peroxiredoxins; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D012313:RNA / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
D004268:DNA-Binding Proteins; D050956:HSP40 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; D056971:Nuclear Receptor Co-Repressor 1; D015513:Oncogene Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D012313:RNA; D000449:Aldehyde Reductase; D007521:Isocitrate Dehydrogenase; D010544:Peroxidases; D054465:Peroxiredoxin VI; D054464:Peroxiredoxins; D017346:Protein Serine-Threonine Kinases; D002401:Cathepsin B