Functional clarification of the bombesin-like peptide system.

铃蟾肽样肽系统的功能澄清。

基本信息

批准号：
12680769
负责人：
OHKI-HAMAZAKI Hiroko
金额：
$ 2.56万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

1. Generation and analysis of mice lacking NMB-R and GRP-R : We obtained male double-knocked out mice as well as male wild-type mice by mating female NMB-R +/-, GRP-R +/- mice and male NMB-R +/- mice. The mutant mice did not show great difference as compared with wild-type mice. The possible alterations observed were the points already noticed in the GRP-R or NMB-R single mutant mice.2. Development of the chick embryos over-expressing NMB-R, GRP-R or BRS-3 : We constructed the retrovirus vectors containing cDNA coding for each subtype of mouse receptpr with EGFP cDNA fused at its C-terminal end. Infection of virus particles prepared using these vectors to the brain region of the chick embryos, resulted in the strong and wide-spread expression of EGFP. We are currently analyzing the phenotypes of these embryos by histological methods.3. Analysis of bombesin systems in birds : In order to elucidate the bombesin like peptides system in birds, we successfully cloned two bombesin-like peptide receptors from chick brain by using RT-PCR with degenerate primers. We named these clones as chGRP-R which showed high similarity with GRP-R of other species, and chBRS-3.5 which showed similarity with BRS-3 and BB4. In situ hybridization analysis showed that chGRP-R was expressed in the hypothalamus and in discrete regions of forebrain, and chBRS-3.5 was widely expressed in the forebrain including striatum. None of these two receptors was expressed in the cerebellum. These results demonstrated that the bombesin-like peptides system exists also in birds and this suggests that this system plays an important role in birds like in mammals.

1。缺乏NMB-R和GRP-R的小鼠的产生和分析：我们通过交配雌性NMB-R +/-，GRP-R +/-小鼠和雄性NMB-R +/-小鼠获得了雄性双敲出小鼠以及雄性野生型小鼠。与野生型小鼠相比，突变小鼠没有显示出很大的差异。观察到的可能改变是在GRP-R或NMB-R单个突变体小鼠中已经注意到的点。2。过表达NMB-R，GRP-R或BRS-3的雏鸡胚胎的开发：我们构建了用EGFP cDNA的每个亚型在其C末端融合的小鼠接收器的cDNA载体。使用这些载体到雏鸡胚胎的大脑区域制备的病毒颗粒的感染导致EGFP的强和广泛表达。我们目前正在通过组织学方法分析这些胚胎的表型3。分析鸟类中的孟买系统：为了阐明鸟类中孟买的肽系统，我们通过将RT-PCR与退化的底漆使用RT-PCR成功克隆了两个bombesin样肽受体。我们将这些克隆命名为CHGRP-R，它与其他物种的GRP-R相似，而CHBRS-3.5与BRS-3和BB4相似。原位杂交分析表明，CHGRP-R在下丘脑和前脑的离散区域中表达，而CHBRS-3.5在包括纹状体在内的前脑中广泛表达。这两个受体都没有在小脑中表达。这些结果表明，鸟类样肽系统也存在于鸟类中，这表明该系统在哺乳动物中的鸟类中起着重要作用。