Characterization of the Placenta Specific Bovine Mammalian Achaete Scute-like Homologue 2 (Mash2) Gene

Mash2, a basic helix-loop-helix transcription factor, stimulates mononucleate trophoblast cell proliferation and inhibits giant/binucleate cell formation. In mice, Mash2 is a maternally expressed imprinted gene. Regulation of bovine Mash2 is unclear due to limited genetic knowledge. Our objectives were to clone and characterize bovine Mash2 and evaluate its imprinting status by utilizing Bos taurus taurus and Bos taurus indicus interspecies crossing. Bovine Mash2 mRNA shares 78% and 70% homology with human and mouse Mash2, with the DNA binding domain (88%) and bHLH region (95%) being highly conserved. Expression of Mash2 mRNA was seen exclusively in cotyledonary areas of the placenta. The greatest abundance of Mash2 mRNA was in day 17 filamentous embryos, during the time of rapid trophoblast proliferation. Reduction in Mash2 mRNA abundance was detected in day 8 parthenogenetic blastocysts suggesting paternal regulation of gene expression. Prior to implantation (days 8 and 17), Mash2 mRNA appears to have biallelic expression, but is paternally silenced after implantation (days 40 and 60). In conclusion, the Mash2 is highly conserved across species and is specifically expressed in the bovine placenta. Bovine Mash2 appears to be maternally expressed after implantation, but the paternal genome plays a role in regulating expression.