Functional analysis of a novel receptor for lysophosphatidic acid (LPA)

溶血磷脂酸（LPA）新型受体的功能分析

• 批准号: 16590219
• 负责人: ISHII Satoshi
• 金额: $ 2.18万
• 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 (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590219/
• 关键词: lysophosphatidic acid; LPA; LPA4; p2y9; GPR23; receptor

B103 rat neuroblastoma cells, which are unable to respond to lysophosphatidic acid (LPA), were stably transfected, with the expression vector for either LPA1 or LPA4 (also known as p2y9/GPR23). I found that, in response to LPA, LPA4 mediated Ca^<2+> responses in B103-LPA4 cells. Furthermore, LPA4 was shown to activate Rho small GTPase, resulting in the morphological changes including cell rounding and cell aggregation. Although B103-LPA1 cells also became rounded upon LPA stimulation, their degree of cell rounding was significantly less than that of B103-LPA4 cells. As for cell aggregation, LPA1 activation did not induce any morphological changes. My observation that, unlike LPA1, LPA4 did not couple to pertussis toxin-sensitive Gi/o proteins may account for these functional differences between LPA1 and LPA4. Because LPA1 plays critical roles in the nervous system such as brain formation and neuropathic pain, my data raise the possibility that LPA4 also may have neuronal functions in vivo.A rabbit polyclonal antiserum was raised against a synthetic peptide derived from mouse LPA4. Actually, LPA4 protein was successfully detected by the antiserum in Western analysis of mouse brain at embryonic day 12, which was rich in LPA4 mRNA, and flow cytometry analysis of the LPA4-expressing B103 cells.The LPA4 gene was disrupted in marine embryonic stem (ES) cells by homologous recombination. Three clones of ES cells carrying the proper homologous recombination events without random integration of the targeting vector were injected into blastocysts. Chimeras from one of the three clones resulted in germline transmission of the targeted allele, which was confirmed by Southern analysis.

对溶血磷脂酸(LPA)不敏感的B103大鼠神经母细胞瘤细胞，用LPA1或LPA4(又称p2y9/GPR23)表达载体稳定转染。我发现，作为对LPA的反应，LPA4介导了B103-LPA4细胞的钙离子反应。此外，LPA4还激活了Rho小GTP酶，导致细胞的形态变化，包括细胞圆形和细胞聚集。虽然B103-LPA1细胞在LPA刺激下也变圆，但其细胞圆化程度明显低于B103-LPA4细胞。在细胞聚集方面，LPA1的激活没有引起细胞形态的改变。我观察到，与LPA1不同，LPA4不与百日咳毒素敏感的Gi/o蛋白偶联，这可能是LPA1和LPA4之间功能差异的原因。由于LPA1在神经系统，如脑形成和神经病理性疼痛中起着关键作用，我的数据增加了LPA4在活体中也可能具有神经元功能的可能性。事实上，在胚胎第12天的小鼠脑组织的Western分析和表达LPA4的B103细胞的流式细胞仪分析中，均能成功检测到LPA4蛋白的表达。将携带适当同源重组事件的三个克隆的ES细胞注射到囊胚中，而不是随机整合靶向载体。来自三个克隆之一的嵌合体导致了目标等位基因的种系传播，这一点得到了Southern分析的证实。

项目成果

Attenuation of folic acid-induced renal inflammatory injury in platelet-activating factor receptor-deficient mice

• DOI: 10.2353/ajpath.2006.050634
• 发表时间: 2006-05-01
• 期刊: AMERICAN JOURNAL OF PATHOLOGY
• 影响因子: 6
• 作者: Doi, K;Okamoto, K;Noiri, E
• 通讯作者: Noiri, E

Platelet-activating factor receptor deficient mice show an unaltered clearance of Haemophilus influenzae from their respiratory tract.

血小板激活因子受体缺陷小鼠的呼吸道中流感嗜血杆菌的清除率没有改变。

• 发表时间: 2004
• 期刊: Shock 22
• 作者: Branger;J.;Wieland;C.W.;Florquin;S.;Maris;N.A.;Pater;J.M.;Speelman;P.;Shimizu;T.;Ishii;S.;van der Poll;T.
• 通讯作者: T.

Identification of T cell death-associated gene 8 ( TDAG8) as a novel acid sensing G-protein-coupled receptor

• DOI: 10.1074/jbc.m407832200
• 发表时间: 2005-03-11
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Ishii, S;Kihara, Y;Shimizu, T
• 通讯作者: Shimizu, T

Platelet activating factor receptor deficient mice have an improved host defense against pneumococcal pneumonia.

血小板激活因子受体缺陷小鼠的宿主对肺炎球菌肺炎的防御能力有所改善。

• 发表时间: 2004
• 期刊: J.Infect.Dis. 189
• 作者: Rijneveld;A.W.;Weijer;S.;Florquin;S.;Speelman;P.;Shimizu;T.;Ishii;S.;van der Poll;T.
• 通讯作者: T.

Absence of platelet-activating factor receptor protects mice from osteoporosis following ovariectomy.

• DOI: 10.1172/jci20504
• 发表时间: 2004-07
• 期刊: The Journal of clinical investigation
• 作者: H. Hikiji;S. Ishii;H. Shindou;T. Takato;Takao Shimizu