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Functional analyses of a seven transmembrane receptor essential for osteoclast differentiation

破骨细胞分化必需的七次跨膜受体的功能分析

基本信息

批准号：
17590253
负责人：
NOMIYAMA Hisayuki
金额：
$ 2.24万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590253/
关键词：
osteoclast seven transmembrane receptor orphan receptor DC--STAMP transmembrane region cDNA membrane fusion cell fusion RAW-D細胞 siRNA スクリーニング酒石酸抵抗性酸性ホスファターゼ

项目摘要

Osteoclasts are bone-resorbing, multinucleated giant cells that are essential for bone remodeling and are formed through cell fusion of mononuclear precursor cells. By cDNA subtraction, we previously isolated a cDNA clone coding for seven-transmembrane receptor DC-STAMP which plays an important role in ostc oclastogenesis (Kukita et al. J. Exp. Med. 200:941, 2004). In order to reveal the function of DC-STAMP, we have tried to identify its ligand using several methods and a mouse cell line which differentiates into osteoclast-like cells upon stimulation. However, positive results have not been obtained so far.Since the zebrafish draft genome sequence does not contain the DC-STAMP gene sequence, we have not been able to do the knock-down experiment. However, we found by database search a novel gene with sequence homology to a portion of the DC-STAMP gene. There are 3 genes containing this DC-STAMP motif in the zebrafish genome. We isolated a cDNA for one of the genes based on the EST data. This gene contains transmembrane regions and may be involved in the cell fusion like DC-STAMP. Recently Drosophila Snky and C. elegans Spe-42 proteins have been shown to have the DC-STAMP motif, although the orthologous relationships with the zebrafish gene are unclear. These transmembrane proteins are shown to be required in egg-sperm membrane fusion. Therefore analyses of the zebrafish genes with the DC-STAMP motifs will provide additional insights into the mechanisms involved in the membrane fusion.

破骨细胞是骨吸收的多核巨细胞，是骨重建所必需的，通过单核前体细胞的细胞融合形成。通过cDNA消减，我们先前分离了编码七跨膜受体DC-STAMP的cDNA克隆，该受体在ostc破细胞发生中起重要作用（Kukita等人，J. Exp. 200：941，2004）。为了揭示DC-STAMP的功能，我们尝试使用几种方法和一种在刺激下分化为破骨细胞样细胞的小鼠细胞系来鉴定其配体。但至今没有得到阳性结果，由于斑马鱼基因组草图序列中不含DC-STAMP基因序列，我们一直没能做敲除实验。然而，我们通过数据库搜索发现了一个新的基因与DC-STAMP基因的一部分序列同源性。在斑马鱼基因组中有3个基因含有这种DC-STAMP基序。我们根据EST数据分离了其中一个基因的cDNA。该基因含有跨膜区，可能与DC-STAMP一样参与细胞融合。最近，果蝇Snky和C.尽管与斑马鱼基因的同源性关系尚不清楚，但已显示秀丽线虫Spe-42蛋白具有DC-STAMP基序。这些跨膜蛋白被证明是卵-精膜融合所必需的。因此，对具有DC-STAMP基序的斑马鱼基因的分析将为膜融合所涉及的机制提供额外的见解。