Relationship between time of first cleavage and the expression of IGF-I growth factor, its receptor, and two housekeeping genes in bovine two-cell embryos and blastocysts produced in vitro.

Abstract

We have previously demonstrated that there is a clear relationship between the time interval between insemination and first cleavage in vitro and the development to the blastocyst stage of bovine embryos. In addition we have shown that this developmental ability can be linked to the stability of the mRNA for several gene transcripts measured in 2-cell bovine embryos cleaving at different times. The aim of this study was to examine the relationship between bovine embryo developmental competence, assessed in terms of time of first cleavage, and the expression of insulin-like growth factor-I (IGF-I) ligand and receptor, hypoxanthine phosphoribosyl transferase (HPRT) and glucose-6-phosphate dehydrogenase (G6PD). The expression of beta-actin was used as a reference value. No differences were observed in the mRNA expression of G6PD and HPRT genes between male and female 2-cell embryos. However, the expression of these two genes was significantly higher in female blastocysts than in male blastocysts. Moreover, when the relative amount of G6PD and HPRT mRNA detected in these groups of male and female embryos was compared, there was a significant relationship between the time of first cleavage and the relative amount of mRNA: 2-cell embryos and blastocysts derived from oocytes that cleaved at 27 and 30 hr post insemination had higher levels of mRNA for G6PD and HPRT than those that cleaved after 33 hr. IGF-I ligand and receptor was detected in all blastocysts analyzed, irrespective of stage of development or time of first cleavage. In addition, the receptor was detected in all 2-cell embryos examined. In contrast, while IGF-I ligand was found in all 2-cell embryos that cleaved at 27 and 30 hpi, it was only found in some of those cleaving between 33 and 36 hpi and in none of those cleaving after 36 hr. In conclusion, we have demonstrated differences in gene expression in the early embryo that are reflective of differences in developmental competence between early- and late-cleaving zygotes.