Ushizawa Koichi, Takahashi Toru, Hosoe Misa, Kizaki Keiichiro, Abe Yasuyuki, Sasada Hiroshi, Sato Eimei, Hashizume Kazuyoshi
Reproductive Biology Research Unit, Division of Animal Sciences, National Institute of Agrobiological Sciences, 2 Ikenodai, Tsukuba, Ibaraki 305-8602, Japan.
BMC Dev Biol. 2007 Mar 15;7:16. doi: 10.1186/1471-213X-7-16.
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. Caprine PRPs are compared with known bovine PRPs. We examined their evolution and role in the ruminant placenta.
Full-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.
We 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.
本研究报告了两种新型山羊催乳素相关蛋白(cPRP1和cPRP6)全长cDNA序列的鉴定,以及它们在胎盘中的定位和定量表达。将山羊PRP与已知的牛PRP进行比较。我们研究了它们在反刍动物胎盘中的进化和作用。
克隆了全长cPRP1和cPRP6 cDNA,其开放阅读框分别为717和720个核苷酸,对应于238和239个氨基酸的蛋白质。cPRP1预测的氨基酸序列与牛PRP1(bPRP1)具有72%的同源性。cPRP6预测的氨基酸序列与牛PRP6(bPRP6)具有74%的同源性。通过RT-PCR仅在胎盘小叶中检测到这两种cPRP以及bPRP。原位杂交分析显示滋养层双核细胞中存在两种cPRP的mRNA。通过实时RT-PCR分析妊娠30、50、90和140天时胎盘小叶中的这些mRNA进行定量。这两个新的cPRP基因都能够在哺乳动物细胞表达系统中翻译出成熟蛋白。蛋白质印迹法确定带有FLAG标签的cPRP1分子大小为33 kDa,带有FLAG标签的cPRP6分子大小为45 kDa。cPRP1和cPRP6的序列特性和定位表达与牛的相似。然而,它们的表达谱与牛胎盘不同。尽管本研究证明了PRP在山羊胎盘中可能的作用,但PRP可能像在牛中一样调节双核细胞功能,但其关键作用仍不清楚。
我们在山羊胎盘中鉴定出了新型PRP。将山羊PRP的定位和定量表达与牛PRP进行了比较。数据表明山羊胎盘PRP基因在妊娠中具有协调功能,就像在牛中一样。这是首次对PRP在山羊胎盘中的功能进行研究。