Differential expression of two genes located on the X chromosome between male and female in vitro-produced bovine embryos at the blastocyst stage

Abstract

The potentially unbalanced expression at preimplantation developmental stages of X-linked genes might be responsible of the faster development of male than female embryos in vitro. Two genes located on the X chromosome, glucose-6-phosphate dehydrogenase (G6PD) and hypoxanthine phosphoribosyl transferase (HPRT), are involved in controlling the amount of oxygen radicals, and hence they might have influence in embryo development. We have quantified mRNA expression of these two genes, using in vitro fertilized-in vitro cultured male and female bovine embryos. In vitro-produced early blastocysts obtained at days 7 and 8 were collected and biopsied for gender determination, and the remaining embryos were kept in LN(2) until RNA purification. After sex determination, embryos were pooled in groups of 3 males or 3 females, and mRNA was purified. Using a semiquantitative sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) assay, we detected G6PD and HPRT mRNA expression at the early blastocyst stage in all bovine embryos analyzed. Moreover, mRNA expression of both genes studied was significantly higher in female embryos than in male embryos. The differential expression of G6PD and HPRT at these early stages confirm that sex differences are evident prior to gonadal differentiation and that preimplantation bovine embryos have sexually dimorphic gene expression at least with respect to G6PD and HPRT transcripts. These differences might be responsible of the faster development in culture of in vitro-produced male bovine that has been reported. Mol. Reprod. Dev. 55:146-151, 2000.