Effect of speed of development on mRNA expression pattern in early bovine embryos cultured in vivo or in vitro.
Recent data have demonstrated that fast-cleaving embryos produced in vitro are more likely to develop to blastocyst stage, and that the postfertilization culture system used impacts considerably on the mRNA expression and quality of blastocysts produced. The present study is the first to investigate the relationship between the developmental speed of embryos produced in vivo or in vitro and the temporal transcription pattern. Genes related to important preimplantation events are monitored during the first 4 days of embryo development in embryos with fast or slow development. The set of genes analyzed in the present study characterizes several important physiological processes including: transport and metabolism of fructose (Glut-5), stress (SOX), mitochondrial activity and detoxification of reactive oxygen species (MnSOD), cell communication (Cx43), maternal recognition of pregnancy (IFN-tau), imprinting (IGF-II), apoptosis (Bax), growth factor binding and metabolism (IGF-IR), and oxidative stress (G6PD). Using real time PCR, we have found that for all the genes analyzed there are differences in mRNA expression between embryos with fast and slow developmental speed produced both in vitro and in vivo. Frequently, genes that may be stress induced such as SOX, MnSOD, BAX, IFtau, and G6PD were highly transcribed in in vitro produced embryos and in embryos with slow developmental speed. On the other side, transcripts from genes related with metabolism, growth, and differentiation (Glut-5, Cx 43, IGF-II, and IGF-IR) were detected in higher amounts in in vivo produced embryos and in embryos with fast developmental speed. Moreover, it is interesting to stand out that for some genetic markers (such as SOX and G6PD) there are in vivo and in vitro differences that can be observed even before materno-zygotic transition, which probably reflects a differential mRNA degradation. These transcription patterns reflects the embryonic response to the adverse in vitro culture conditions, and connect the low quality of embryos which slow developmental speed produced in vivo and in vitro, with the mRNA expression pattern of some embryonic genes.
Gutiérrez-Adán
A
A
Dpto. de Reproducción Animal y Conservación de Recursos Zoogenéticos, INIA, Madrid, Spain. agutierr@inia.es
Rizos
D
D
Fair
T
T
Moreira
P N
PN
Pintado
B
B
de la Fuente
J
J
Boland
M P
MP
Lonergan
P
P
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002417:Cattle; D017931:DNA Primers; D047108:Embryonic Development / Q000502:physiology*; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D009924:Organ Culture Techniques; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015053:Zygote / Q000502:physiology
D017931:DNA Primers; D011506:Proteins; D012333:RNA, Messenger