Studies on the placental PRL-like molecules and their target cells

胎盘PRL样分子及其靶细胞的研究

• 批准号: 10460121
• 负责人: SHIOTA Kunio
• 金额: $ 8.51万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10460121/
• 关键词: rat; placenta; pregnancy; PRL; PRL-H; SSP; PL-Im; baculovirus; PLP; PLP-H

The placenta acts as a multi-function organ in the maintenance of pregnancy and in fetal growth. Rat placenta produces several members of the placental prolactin (PRL) family that show structural similarities to pituitary PRL. To date more than ten members of the placental PRL family have been cloned, and they are categorized into two subfamilies according to their Structure and function; the placental lactogen (PL) subfamily and like protein (PLP) subfamily. PRL exerts its biological function through the PRL receptor (PRL-R). PL subfamily members are also able to bind to the PRL-R and to stimulate the proliferation of Nb2 cells. Some members of PLP sub family do not bind to PRL-R nor do they stimulate lactogenic activity even though they are structura11 similar to PRL, implying that there may be yet unknown specific receptors for the PRL family members. To further understand the mechanisms underlying the expression of biological activities of PRL family members, we first evaluated the biological activity of recombinant PL-Im (recPL-Im) produced by means of a baculovirus/insect cell expression system The recPL-Im showed not only an ability to bind to PRL-R and express PRL-like activity, demonstrated by the Nb2 cell proliferation assay, but also showed luteotropic hormone (LH)-like activity in vitro. We then evaluated the role of N-linked oligosaccharides in the mo1ecular action of recPL-Im and found that the ability of PL-Im to bind PRL receptor is independent of the N-glycosylation although it appeared to be relevant for expressing PRL-like activity in the Nb2 cell proliferation assay. These results raised the possibility that recPL-Im may also act through unidentified receptor system besides the PRL-R. In this study we also performed of cDNAs for rat placenta-specific cytokines/ hormones, and isolated a novel member of PLP subfamily, named PLP-H, as well as a novel secretory protein, SSP.

胎盘作为维持妊娠和胎儿生长的多功能器官。大鼠胎盘产生胎盘催乳素 (PRL) 家族的几个成员，其结构与垂体 PRL 相似。迄今为止，胎盘PRL家族已有十多个成员被克隆，根据其结构和功能分为两个亚家族：胎盘催乳素 (PL) 亚家族和类似蛋白 (PLP) 亚家族。 PRL通过PRL受体（PRL-R）发挥其生物学功能。 PL 亚家族成员还能够与 PRL-R 结合并刺激 Nb2 细胞的增殖。 PLP亚家族的一些成员不与PRL-R结合，也不刺激泌乳活性，尽管它们的结构与PRL相似，这意味着PRL家族成员可能还存在未知的特异性受体。为了进一步了解 PRL 家族成员生物活性表达的机制，我们首先评估了通过杆状病毒/昆虫细胞表达系统产生的重组 PL-Im (recPL-Im) 的生物活性。recPL-Im 不仅表现出与 PRL-R 结合并表达 PRL 样活性的能力（Nb2 细胞增殖试验证明），而且还表现出促黄体激素 (LH) 样活性。 体外。然后我们评估了 N-连接寡糖在 recPL-Im 分子作用中的作用，发现 PL-Im 结合 PRL 受体的能力独立于 N-糖基化，尽管它似乎与 Nb2 细胞增殖测定中表达 PRL 样活性相关。这些结果提出了除了 PRL-R 之外，recPL-Im 还可能通过未识别的受体系统发挥作用的可能性。在这项研究中，我们还对大鼠胎盘特异性细胞因子/激素进行了 cDNA 分析，并分离了 PLP 亚家族的一个新成员，命名为 PLP-H，以及一个新的分泌蛋白，SSP。

项目成果

K.Iwatsuki et al.: "A novel secretory protein by rat spongiotrophoblast"Biology of Reproduction. (in press). (2000)

K.Iwatsuki 等人：“大鼠海绵滋养层的新型分泌蛋白”生殖生物学。

Iwatsuki, K. et al.: "Molecular cloning and characterization of a new member of the rat placental prolactin (PRL) family, PRL-like protein H"Endocrinology. 139. 4976-83 (1998)

Iwatsuki, K. 等人：“大鼠胎盘催乳素 (PRL) 家族新成员 PRL 样蛋白 H 的分子克隆和表征”内分泌学。

Hirosawa-Takamori, M. et al.: "Characterization of rat mid-pregnancy-specific placental lactogen produced by baculovirus/insect cell expression system."J Reprod Dev. 45. 43-50 (1999)

Hirosawa-Takamori, M. 等人：“杆状病毒/昆虫细胞表达系统产生的大鼠妊娠中期特异性胎盘催乳素的表征。”J Reprod Dev。

Iwatsuki, K. et al.: "A novel secretory protein produced by rat spongiotrophoblast."Biology of Reproduction. (in press). (2000)

Iwatsuki, K. 等人：“大鼠海绵滋养层产生的一种新型分泌蛋白。”生殖生物学。

Iwatsuki, K. et al.: "A novel secretory protein produced by rat spongiotrophoblast"Biology of Reproduction. (in press). (2000)

Iwatsuki, K. 等人：“大鼠海绵滋养层产生的一种新型分泌蛋白”生殖生物学。