Suppressed expression of genes involved in transcription and translation in in vitro compared with in vivo cultured bovine embryos.
In vivo-derived bovine embryos are of higher quality than those derived in vitro. Many of the differences in quality can be related to culture environment-induced changes in mRNA abundance. The aim of this study was to identify a range of mRNA transcripts that are differentially expressed between bovine blastocysts derived from in vitro versus in vivo culture. Microarray (BOTL5) comparison between in vivo- and in vitro-cultured bovine blastocysts identified 384 genes and expressed sequence tags (ESTs) that were differentially expressed; 85% of these were down-regulated in in vitro cultured blastocysts, showing a much reduced overall level of mRNA expression in in vitro- compared with in vivo-cultured blastocysts. Relative expression of 16 out of 23 (70%) differentially expressed genes (according to P value) were verified in new pools of in vivo- and in vitro-cultured blastocysts, using quantitative real-time PCR. Most (10 out of 16) are involved in transcription and translation events, suggesting that the reason why in vitro-derived embryos are of inferior quality compared with in vivo-derived embryos is due to a deficiency of the machinery associated with transcription and translation.
Corcoran
D
D
School of Agriculture, Food Science and Veterinary Medicine and Centre for Integrative Biology, Conway Institute for Biomolecular and Biomedical Research, College of Life Sciences, University College Dublin, Belfield, Dublin 4, Ireland.
Fair
T
T
Park
S
S
Rizos
D
D
Patel
O V
OV
Smith
G W
GW
Coussens
P M
PM
Ireland
J J
JJ
Boland
M P
MP
Evans
A C O
AC
Lonergan
P
P
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000502:physiology*; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D047108:Embryonic Development / Q000235:genetics*; D020224:Expressed Sequence Tags; D005260:Female; D005307:Fertilization in Vitro; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization / Q000379:methods; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D011247:Pregnancy; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D017931:DNA Primers; D012333:RNA, Messenger