Evaluation of the quality of porcine somatic cell nuclear transfer embryo by gene transcription profiles.
This study aimed to evaluate the quality of porcine somatic cell nuclear transfer (SCNT) embryos by examining its gene transcription patterns. Embryos were produced by SCNT, intracytoplasmic sperm injection (ICSI) or under different conditions, and transcripts of genes for fibroblast growth factor receptor (FGFr) 2IIIc, FGFr72IIIb, X inactive-specific transcript (Xist), interleukin 6 (IL6), IL6 receptor (IL6r) alpha and c-kit ligand, were detected by real-time RT-PCR. The percentages of embryos in which these transcripts were detected were similar in SCNT and ICSI embryos. On the other hand, the transcriptional levels of the FGFr72IIIb and IL6ralpha genes were 0.5 times less and 2 times more, respectively, in SCNT blastocysts than those of ICSI blastocysts (p<0.05). When nuclear transfer was performed before or after activation of oocytes, embryos in the latter case showed significantly lower frequencies of having FGFr72IIIb (74% vs. 90%) and Xist (3% vs. 33%) transcripts compared to the former case embryos (p<0.05). When two lines of nuclear donor cells with different developmental potencies were used, the transcriptional profiles in the reconstructed embryos did not show any significant differences. Our study suggests that expression profiles of FGFr72IIIb, IL6ralpha, and Xist can be used as markers for the diagnosis of the developmental potency of porcine nuclear transfer embryos.
Miyazaki
Koji
K
R&D Laboratory, Nipro Corporation, Shiga, Japan.
Tomii
Ryo
R
Kurome
Mayuko
M
Ueda
Hideto
H
Hirakawa
Kazumasa
K
Ueno
Satoshi
S
Hiruma
Katsumi
K
Nagashima
Hiroshi
H
D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D019976:Cloning, Organism; D017931:DNA Primers / Q000737:chemistry; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000378:metabolism; D047108:Embryonic Development; D004628:Embryonic Structures / Q000378:metabolism; D005307:Fertilization in Vitro; D009865:Oocytes / Q000378:metabolism; D012313:RNA / Q000737:chemistry; D027724:Reproductive Techniques, Assisted; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods; D013552:Swine; D013997:Time Factors; D014158:Transcription, Genetic
D017931:DNA Primers; D012313:RNA