Gene expression profiles of novel caprine placental prolactin-related proteins similar to bovine placental prolactin-related proteins
Gene expression profiles of novel caprine placental prolactin-related proteins similar to bovine placental prolactin-related proteins
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DOI：
10.1186/1471-213x-7-16
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发表时间：
2007-03-15
影响因子：
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通讯作者：
Hashizume, Kazuyoshi
中科院分区：
文献类型：
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作者：
Ushizawa, Koichi;Takahashi, Toru;Hashizume, Kazuyoshi
背景：本研究克隆了两个新的山羊催乳素相关蛋白(cPRP1和cPRP6)的全长cDNA序列，并对其在胎盘中的定位和定量表达进行了研究。并与已知的牛PRPS进行了比较。结果：克隆到全长的cPRP1和cPRP6基因，其开放阅读框分别为717和720个核苷酸，分别对应于238和239个氨基酸的蛋白质。所预测的氨基酸序列与牛PRP1(BPRP1)的同源性为72%。CPRP6的氨基酸序列与牛PRP6(BPRP6)的同源性为74%。RT-PCR仅在胎盘中检测到这两个cPRPs和bprp。原位杂交分析表明，在滋养层双核细胞中均存在cPRPs基因的表达。在妊娠30天、50天、90天和140天对胎盘组织进行实时RT-PCR分析，以定量检测这些基因的表达。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成熟的蛋白质。Western blotting确定带有标志标签的cPRP1的分子大小为33 kDa，带有标志标签的cPRP6的分子大小为45 kDa。CPRP1和cPRP6的序列性质和定位表达与牛相似。然而，它们在牛胎盘中的表达谱不同。虽然本研究证实了PRPS在山羊胎盘中的可能作用，但PRPS可能像牛一样调节双核细胞的功能，但其关键作用尚不清楚。结论：我们在山羊胎盘中发现了新的PRPS。比较了山羊PRPS和牛PRPS的定位和定量表达。这些数据表明，山羊胎盘中的PRP基因与牛胎盘中的PRP基因一样，对妊娠具有协调功能。这是首次对山羊胎盘中PRPS功能的研究。
Background: 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.Results: 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.Conclusion: 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.