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Background  Bovine trophoblast binucleate cells (BNC) express a plethora of molecules including bovine placental lactogen (bPL, gene name is  bCSH1 ) and bovine prolactin-related protein-1 ( bPRP1 ).  BCSH1  and  bPRP1  are members of the growth hormone (GH)/prolactin (PRL) gene family, which are expressed simultaneously in BNC and are central to placentation and the progression of pregnancy in cattle. However, there is a paucity of information on the transcriptional regulatory mechanisms of both the  bCSH1  and  bPRP1  genes. Recent studies, however, have demonstrated that the expression of a number of genes is controlled by the methylation status of their promoter region. In the present study, we examined the cell-type-specific epigenetic alterations of the 5'-flanking region of the  bCSH1  and  bPRP1  genes to gain an insight into their regulatory mechanisms.    Results  Analysis of 5-aza-2'-deoxycytidine treatment demonstrated that  bCSH1  expression is moderately induced in fibroblast cultures but enhanced in BT-1 cells. Sodium bisulfite based sequencing revealed that  bCSH1  is hypomethylated in the cotyledonary tissue but not in the fetal skin, and this pattern was not altered with the progression of pregnancy. On the other hand, the methylation status of  bPRP1  was similar between the cotyledon and fetal skin. The  bPRP1  gene was exclusively hypermethylated in a bovine trophoblast cell-derived BT-1 cell-line. While the activity of  bCSH1  was similar in both BT-1 and bovine fibroblast cells, that of  bPRP1  was specific to BT-1. Treatment with a demethylating agent and luciferase assays provided in vitro evidence of the positive regulation of  bCSH1  but not  bPRP1 .    Conclusion  This is the first report to identify the differential regulatory mechanisms of the  bCSH1  and  bPRP1  genes and indicates that  bCSH1  might potentially be the only transcript that is subject to DNA methyltransferase regulation. The data indicates the possibility of novel kinetics of induction of the synchronously expressed BNC-specific  bCSH1  and  bPRP1  transcripts, which may aid the understanding of the intricate regulation and specific role(s) of these important molecules in bovine placentogenesis and the progression of pregnancy.

The characterization of DNA methylation-mediated regulation of bovine placental lactogen and bovine prolactin-related protein-1 genes