Effect of speed of development on mRNA expression pattern in early bovine embryos cultured in vivo or in vitro

Abstract

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.

Increasing of blastocyst rate and gene expression in co-culture of bovine embryos with adult adipose tissue-derived mesenchymal stem cells

“…One of the genes positively influenced by co-culture with MSCs was POU5F1 which is considered a transcriptional regulator necessary for the maintenance of pluripotency in the developing embryo and embryonic stem cells of mammals [34,35]. Similarly, we also observed an increase in G6PDH expression, a gene related to carbohydrate metabolism and NADPH availability for steroid and fatty acid biosynthesis, as well as responsible for increasing the ribose 5-phosphate supply for nucleotide and nucleic acid synthesis [36][37][38]. Both genes are considered good markers of embryo quality short of the possibility of performing transfer and evaluating postimplantation development [39][40][41][42][43][44].…”

Resveratrol treatment during goat oocytes maturation enhances developmental competence of parthenogenetic and hand-made cloned blastocysts by modulating intracellular glutathione level and embryonic gene expression

“…In case of HMC-derived blastocysts reconstructed from mature oocytes of different resveratrol treatment groups, expression of a set of genes which are having relevance in early embryonic development [13,25,35,42,43] were checked. The genes included in the present study are involved in pre-and postnatal growth (IGF -1 ), metabolism (Glut1 ), pluripotency (Oct4), morphogenesis and biogenesis of the trophoectoderm (E -cad), lineage determination and cavitation (Stat3), gap junction, cavitation and cryosurvival (Cx43), growth arrest (CHOP-10), oxidative stress (MnSOD), and DNA methylation (DNMT).…”

Temporal detection of caspase-3 and -7 in bovine in vitro produced embryos of different developmental capacity

“…Fast cleaving bovine embryos have an altered gene expression (Lonergan et al 2000, Ward et al 2001, Gutiérrez-Adán et al 2004, Dode et al 2006 or altered polyadenylation status of several developmentally important gene transcripts (Pocar et al 2001, Brevini-Gandolfi et al 2002 and a higher chance of reaching advanced developmental stages in comparison with late cleaving embryos (Van Soom et al 1992, Dinnyés et al 1999, Lequarré et al 2003, Favetta et al 2004, Gutiérrez-Adán et al 2004. Studies investigating the effect of developmental kinetics on the incidence of apoptosis during preimplantation embryo development in bovine are scarce.…”