Abstract
BackgroundThis study reports the identification of a full-length cDNA sequence for two novel caprine prolactin-related proteins (cPRP1 and cPRP6), and their localization and quantitative expression in the placenta. Caprine PRPs are compared with known bovine PRPs. We examined their evolution and role in the ruminant placenta.ResultsFull-length cPRP1 and cPRP6 cDNA were cloned with a 717- and 720- nucleotide open-reading frame corresponding to proteins of 238 and 239 amino acids. The cPRP1 predicted amino acid sequence shares a 72% homology with bovine PRP1 (bPRP1). The cPRP6 predicted amino acid sequence shares a 74% homology with bovine PRP6 (bPRP6). The two cPRPs as well as bPRPs were detected only in the placentome by RT-PCR. Analysis by in situ hybridization revealed the presence of both cPRPs mRNA in the trophoblast binucleate cells. These mRNA were quantified by real-time RT-PCR analysis of the placentome at 30, 50, 90 and 140 days of pregnancy. Both new cPRP genes were able to translate a mature protein in a mammalian cell-expression system. Western blotting established the molecular sizes of 33 kDa for cPRP1 with FLAG-tag and 45 kDa for cPRP6 with FLAG-tag. The sequence properties and localized expression of cPRP1 and cPRP6 were similar to those of bovine. However, their expression profiles differed from those in bovine placenta. Although this study demonstrated possible roles of PRPs in caprine placenta, PRPs may regulate binucleate-cell functions like those in bovine, but their crucial roles are still unclear.ConclusionWe have identified the novel PRPs in caprine placenta. Localization and quantitative expression of caprine PRPs were compared with bovine PRPs. The data indicate that PRP genes in caprine placenta have coordination functions for gestation, as they do in bovine. This is the first study of PRPs function in caprine placenta.
Highlights
This study reports the identification of a full-length cDNA sequence for two novel caprine prolactin-related proteins, and their localization and quantitative expression in the placenta
Sequences of caprine prolactin-related protein1 (cPRP1) and caprine prolactin-related protein-6 (cPRP6) cDNA and deduced amino acids Sequences of 933 and 957 nucleotides were isolated from caprine placentome and cloned as candidates of cPRP1 and cPRP6
The amino acid sequences deduced from full-length cPRP1 and cPRP6 cDNA are 238 and 239 amino acids
Summary
This study reports the identification of a full-length cDNA sequence for two novel caprine prolactin-related proteins (cPRP1 and cPRP6), and their localization and quantitative expression in the placenta. Various kinds of functional molecules such as steroid hormones, peptides, and prostanoids are expressed. These molecules contribute to a successful pregnancy and to the establishment of placenta. This paper introduces two novel types of cPRP genes that emerged from the cloning of a complete sequence of full-length cDNA and its expression. We named these sequences caprine prolactin-related protein (cPRP1) and caprine prolactin-related protein-6 (cPRP6), based on their similarity to bovine PRPs
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