A comparative expression analysis of gene transcripts in post-fertilization developmental stages of bovine embryos produced in vitro or in vivo.
This study was carried out to examine the temporal variation in the relative abundance of transcripts during the post-fertilization stages of bovine embryos derived from in vitro or in vivo culture. For this purpose, cumulus-oocyte complexes obtained from ovaries from slaughterhouses were matured, fertilized and cultured in vitro. The in vitro culture was carried out using CR1 medium. In vivo embryos were derived from superovulated and artificially inseminated Simmental heifers and cows. Embryos were recovered from both systems approximately at day 1 (two-cell), day 2 (four-cell), day 3 (eight-cell), day 4 (16-cell), day 5 (morula) and day 7 (blastocyst) after insemination. Relative abundance of target transcripts was performed using real-time PCR. The transcripts studied were the nucleosome assembly protein, mRNA for alivin 1, Pleckstrin homology Sec7 coiled domain, polyadenylate binding protein, NADH dehydrogenase subunit 2, high-mobility group transcription factor, cytokine-like nuclear factor, NY-REN-58 antigen mRNA, mRNA for KIAA1764 and one novel transcript. These transcripts were derived from our previous study conducted using mRNA differential display reverse transcription-polymerase chain reaction (DDRT-PCR) between in vitro-produced bovine eight-, 16-cell, morula and blastocyst-stage embryos. The results of the present study have demonstrated the temporal variation in the relative abundance of these transcripts between in vitro and in vivo culture systems. Such variation in the relative abundance of transcripts during the post-fertilization developmental stages of in vitro and in vivo embryos may reflect the effect of the in vitro culture condition on the transcriptional activity thereby on the developmental competence of the resulting embryos.
Tesfaye
D
D
Institute of Animal Breeding Science, University of Bonn, Bonn, Germany. tesfaye@itz.uni-bonn.de
Ponsuksili
S
S
Wimmers
K
K
Gilles
M
M
Schellander
K
K
D000818:Animals; D002417:Cattle / Q000196:embryology*; D047108:Embryonic Development / Q000235:genetics*; D005260:Female; D005306:Fertilization; D005307:Fertilization in Vitro / Q000662:veterinary*; D015870:Gene Expression; D007315:Insemination, Artificial / Q000662:veterinary; D009419:Nerve Tissue Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D013480:Superovulation; D013997:Time Factors; D046509:Tissue Culture Techniques; D020858:Tissue and Organ Harvesting / Q000662:veterinary; D014157:Transcription Factors / Q000235:genetics
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D014157:Transcription Factors