Temporal expression of transcripts related to embryo quality in bovine embryos cultured from the two-cell to blastocyst stage in vitro or in vivo.
The post-fertilization embryo culture environment can have a dramatic effect on the pattern of gene expression in the embryo and it is widely acknowledged that bovine embryos derived from in vitro culture are of inferior quality to those derived in vivo. The objective of this study was to examine temporal variation in the mRNA abundance of several transcription and translation factors known to differ between blastocysts produced following culture in vitro and in vivo. Embryos were recovered from two in vitro culture systems SOF1 or SOF2 at five developmental stages: 2- to 4-cell, 8-cell, 16-cell, morula, and blastocyst. In vivo embryos were produced from superovulated and artificially inseminated heifers and recovered at approximately 40 hr or 3, 4, 5, and 7 days postinsemination. Blastocysts were also produced following in vitro maturation, in vitro fertilization and culture in the ewe oviduct. Analysis of relative transcript abundance for FOXO3A, EEF1G, HMG2, and REA was performed using quantitative real-time PCR. Irrespective of culture environment each transcript followed, approximately the same general pattern of expression where relative abundance decreased dramatically from the 2- to 4-cell stage to 8-cell stage and increased from the morula to blastocyst stage (P < 0.05). Transcripts for GNBL2 were not observed between the 2- and 16-cell stage of development. Relatively high expression at the 2- to 4-cell indicated that these transcripts are most likely of maternal origin produced in the oocyte during growth and final maturation. A culture-induced change in mRNA abundance of transcription and translation factors was evident in embryos that were produced not only between in vivo and in vitro culture environments but also between different in vitro culture systems.
Corcoran
Deirdre
D
School of Agriculture, Food Science and Veterinary Medicine, Conway Institute for Biomolecular and Biomedical Research, College of Life Sciences, University College Dublin, Belfield, Dublin, Ireland.
Rizos
Dimitrios
D
Fair
Trudee
T
Evans
Alex C O
AC
Lonergan
Patrick
P
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D005260:Female; D018507:Gene Expression Regulation, Developmental