Identification of maternal mRNAs in porcine parthenotes at the 2-cell stage: a comparison with the blastocyst stage.
Successful embryonic development is dependent on the temporal and stage-specific expression of appropriate genes. Currently, information on specific gene expression during early cleavage-stage embryos before zygotic gene activation (ZGA) is limited. In the present study, we compare gene expression between porcine 2-cell and blastocyst stage parthenotes to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based GeneFishing PCR. Using 60 ACPs, we identified and sequenced nine differentially expressed genes (DEGs). A BLAST search revealed that cloned genes or ESTs (GDI-2, MTMR3, MKLN1, NUP88, ePAD, CIRHIM, UPF3B, ITGA2, and CGI-140) had significant sequence similarities with known genes (78-95%) of other species in the GenBank/EMBL database. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) data disclosed that these genes were regulated upstream in metaphase II (MII) oocyte, 1-cell, and 2-cell stage embryos during early pre-implantation. Similarly, upregulation was observed in MII mouse oocytes and 1-cell stage embryos before ZGA, suggesting that these nine differentially expressed orthologous genes play important roles during early cleavage before ZGA. Further analysis of the differentially expressed genes identified in this report should provide the basis for research on early cleavage and activation of the embryonic genome.
Hwang
Kyu-Chan
KC
Department of Animal Sciences, Chungbuk National University, Gaesin-dong, Cheongju, Chungbuk 361-763, South Korea.
Lee
Hwa-Young
HY
Cui
Xiang-Shun
XS
Kim
Jin-Hoi
JH
Kim
Nam Hyung
NH
D000818:Animals; D001483:Base Sequence; D019295:Computational Biology; D017931:DNA Primers; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005796:Genes / Q000235:genetics; D016014:Linear Models; D008431:Maternal-Fetal Exchange / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D010312:Parthenogenesis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D034421:Sus scrofa / Q000196:embryology / Q000378:metabolism*; D015533:Transcriptional Activation
D017931:DNA Primers; D012333:RNA, Messenger