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Development of Central Neurons System and placental fuction.

中枢神经元系统和胎盘功能的发育。

基本信息

批准号：
07306024
负责人：
SHIOTA Kunio
金额：
$ 2.37万
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 (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

Development of various organs begins immediately after initiation of placentation. We previously showed that rat amniotic fluid contains immunoreactive placental lactogen-like molecules with different molecular sizes and pI values from the authentic placental lactogens (PLs) in the rat. In the present study, immunoreactive PLs residing in the amniotic fluid were characterized further by two-dimensional SDS gel electrophoresis and we confirmed that amniotic fluid collected from rats on day 12 of pregnancy contains two PL-like molecules, tentatively called A1 (MW 75 kDa, pI4.6) and A2 (MW 99-102 kDa, pI 5.3-5.4). PLs have ability to bind to prolactin (PRL) receptor and one of the target tissues for PLs is the brain in adult animals. Therefore, we tried to detect PRL receptor (PRL-R) mRNAs in fetal rat brains, and the gene expression is detected very early in developing fetal brain. To evaluate a role for N-linked oligosaccharides in the molecular action of rat placental lactogen-I mosaic … More (PL-Im), recombinant PL-Im (recPL-Im) and three recPL-Im mutants were produced in COS7 cells. It became evident that PL-Im was N-glycosylated at both Asn79 and Asn128. In addition, the two N-linked oligosaccharides which are involved in its biological activity. Interestingly, the ability of PL-Im to bind PRL-R and activate JAK2 appears to be independent of the N-glycosylation. We also found a novel gene, 4P16, which is related to the proliferation of neural cells. The gene expression is also stimulated by PL-Im in the Nb2 cell line. With the aim of identifying molecules that are expressed specifically in the brain during neurogenesis, we tried to generate monoclonal antibodies which recognize molecules showing unique temporal expression patterns and molecular characteristics. One of the antibodies, Mab3C8, recognized a 100-kDa antigen that is enriched in fetal brain. This 100-kDa antigen was constantly expressed during fetal life (from E12 to E20) and became undetectable two days after birth. The molecule unique to the fetal brain, an O-linked sialoglycoprotein with a molecular weight of 100 kDa (FOG100), was found by generating an antibody. In addition to these results, we also found that the fetal brain express the mRNAs for tree orphan receptors, NGFI-B,Rev-erbAa, Rev-erbAb. Finally, we have idenstified several forms of G protein beta sabunit with different PI values in the placenta and brain. Less

各种器官的发育在胎盘形成开始后立即开始。我们以前表明，大鼠羊水中含有免疫反应性胎盘催乳素样分子，具有不同的分子大小和pI值从真正的胎盘催乳素（PL）在大鼠。在本研究中，免疫反应性PLs驻留在羊水的特征进一步通过二维SDS凝胶电泳，我们证实，从大鼠妊娠第12天收集的羊水含有两个PL样分子，暂时称为A1（分子量75 kDa，pI4.6）和A2（分子量99-102 kDa，pI5.3 -5.4）。PL具有与催乳素（PRL）受体结合的能力，并且PL的靶组织之一是成年动物的脑。因此，我们试图检测胎鼠脑中的PRL受体（PRL-R）mRNA，该基因在发育中的胎脑中很早就被检测到。评价N-连接寡糖在大鼠胎盘催乳素-I嵌合体分子作用中的作用 ...更多信息在C 0 S7细胞中产生重组PL-Im（recPL-Im）、重组PL-Im（recPL-Im）和三种recPL-Im突变体。很明显，PL-Im在Asn 79和Asn 128处均被N-糖基化。此外，这两种N-连接的寡糖也参与其生物活性。有趣的是，PL-Im结合PRL-R和激活JAK 2的能力似乎与N-糖基化无关。我们还发现了一个与神经细胞增殖相关的新基因4P 16。在Nb 2细胞系中，PL-Im也刺激基因表达。为了鉴定在神经发生过程中在大脑中特异性表达的分子，我们试图产生识别显示独特的时间表达模式和分子特征的分子的单克隆抗体。其中一种抗体Mab 3C 8识别在胎儿脑中富集的100 kDa抗原。这种100-kDa抗原在胎儿期（从E12到E20）持续表达，出生后两天检测不到。通过产生抗体发现了胎儿脑特有的分子，分子量为100 kDa的O-连接唾液酸糖蛋白（FOG 100）。除此之外，我们还发现胎脑表达三种孤儿受体NGFI-B、Rev-erbAa、Rev-erbAb的mRNA。最后，我们已经阐明了几种形式的G蛋白β亚单位与不同的PI值在胎盘和大脑。少