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

胎盘启动后，各种器官的开发立即开始。我们先前表明，大鼠羊水含有来自大鼠真实的lopenal乳糖量（PLS）的不同分子大小和PI值的免疫反应性位置类乳酸分子样分子。 In the presentation 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, tenantly called A1 (MW 75 kDa, pI4.6) and A2 (MW 99-102 kDa, pI 5.3-5.4). PL具有与催乳素（PRL）受体结合的能力，而PLS的目标时间之一是成年动物的大脑。因此，我们试图检测胎儿大鼠大脑中的PRL受体（PRL-R）mRNA，并且在发育中很早就检测到基因表达。胎儿大脑。为了评估N-连接的寡糖在大鼠lopenal乳酸乳元I-I马赛克的分子作用中的作用……更多（PL-IM），重组PL-IM（RECPL-IM）和三个RecpL-IM突变体在COS7细胞中产生。证据表明，PL-IM在ASN79和ASN128中均被N-糖基化。此外，与其生物活性有关的两种N连接的寡糖。有趣的是，PL-IM结合PRL-R和激活JAK2的能力似乎与N-糖基化无关。我们还发现了一个新型基因4p16，该基因与神经元细胞的增殖有关。 NB2细胞系中的PL-IM也刺激了基因表达。为了鉴定神经发生过程中在大脑中特异性表达的分子，我们试图产生单克隆抗体，这些抗体识别显示出独特的临时表达模式和分子特征的分子。其中一种抗体MAB3C8识别出富含胎儿大脑的100 kDa抗原。这种100 kDa的抗原在胎儿寿命（从E12到E20）中一直表达，并在出生两天后变得无法检测到。通过产生抗体，可以发现胎儿脑特有的分子，一种分子量为100 kDa（FOG100）的O连接的唾液酸糖蛋白（FOG100）。除这些结果外，我们还发现，胎儿大脑对树孤儿受体（NGFI-B，Rev-Erbaa，Rev-erbab）表示mRNA。最后，我们幻想了几种形式的G蛋白βsabunit，其plapeta和脑中具有不同的PI值。较少的