Neural recognition molecules, NrCAM and Contactin, during development and aging.

发育和衰老过程中的神经识别分子 NrCAM 和 Contactin。

基本信息

批准号：
10832010
负责人：
HOSOYA Hiroko
金额：
$ 2.11万
依托单位：
Tokyo Metropolitan Institute of Gerontology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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-10832010/
关键词：
NrCAM contactin adhesion molecule splicing isoform Ig-superfamily Nr-CAM 糖鎖脳の発達接着分子

项目摘要

Various cell adhesion molecules participate in the cell recognition events not only during development, but also in adulthood and during aging. We have been studying the biological roles of contactin, a neural adhesion molecule in the immunoglobulin superfamily, in the aged brain. NrCAM is one of the immunoglobulin superfamily molecules, which is known to interact with contactin and TAG-1 of the contactin subgroup. In this study, we first investigated the expression of NrCAM during development and aging. In the Western blot analysis, we found two bands of 135-kDa and 130-kDa in the adult rat brain. The 130-kDa band was not detected until postnatal day 10, whereas the 135-kDa band was detected from embryo to adulthood. The cDNA that we found encoded a splicing isoform of NrCAM, which has additional 19 amino acid residues in the N-terminal region comparing with the NrCAM reported previously. In order to know whether the two bands on the Western blot analysis were the splicing isoforms or not, we performed RT-PCR. The developmental expression pattern of these two isoform transcripts detected by the RT-PCR conformed to the appearance of the two bands on the Western blot. These results have suggested that the two splicing isoforms play different roles form each other in the brain function. Next, we examined the interaction of NrCAM with the other contactin subgroup molecules. By the immunoprecipitation, it was demonstrated that NrCAM interacted with contactin and TAG-1 in the hippocampus, whereas the interaction with NB-3 of the another contactin subgroup molecule was not detected. Taken together, NrCAM do not interact with all the contactin subgroup molecules. The results obtained here will be of great use for understanding the roles of NrCAM during aging.

各种细胞粘附分子不仅在发育过程中，而且在成年期和衰老期间都参与细胞识别事件。我们一直在研究接触蛋白的生物学作用，这是衰老大脑中免疫球蛋白超家族中神经粘附分子的生物学作用。 NRCAM是免疫球蛋白超家族分子之一，已知与接触蛋白亚组的接触蛋白和TAG-1相互作用。在这项研究中，我们首先研究了在开发和衰老过程中NRCAM的表达。在Western印迹分析中，我们在成年大鼠大脑中发现了两个135 kDa和130 kDa的频段。直到产后第10天才检测到130 kDa频段，而从胚胎到成年后，检测到135 kDa频段。我们发现的cDNA编码了NRCAM的剪接同工型，在N末端区域中还有19个氨基酸残基与先前报道的NRCAM相比。为了知道Western印迹分析上的两个频段是否是剪接同工型，我们进行了RT-PCR。 RT-PCR检测到的这两个同工型转录本的发育表达模式符合蛋白质印迹上两个频段的外观。这些结果表明，两种剪接同工型在大脑功能中彼此之间起着不同的作用。接下来，我们检查了NRCAM与其他接触蛋白亚组分子的相互作用。通过免疫沉淀，证明NRCAM与海马中的接触蛋白和TAG-1相互作用，而与另一个接触素亚组分子的NB-3相互作用。综上所述，NRCAM并未与所有接触蛋白亚组分子相互作用。此处获得的结果将极大地用于理解NRCAM在衰老过程中的作用。