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

• Project/Area Number: 16590219
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: The University of Tokyo
• Principal Investigator: ISHII Satoshi The University of Tokyo, Graduate School of Medicine, Lecturer, 大学院・医学系研究科, 講師 (10300815)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,400,000 (Direct Cost: ¥3,400,000); Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: lysophosphatidic acid / LPA / LPA4 / p2y9 / GPR23 / receptor

Research Abstract

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.

Research Products

• [Journal Article] Involvement of the platelet activating factor receptor in host defense against Streptococcus pneumoniae during postinfluenza pneumonia. (2006)
  • Author(s): van der Sluijs, K.F., van Elden, L.J.R., Nijhuis, M., Schuurman, R., Florquin, S., Shimizu, T., Ishii, S., Jansen, H.M., Lutter, R., van der Poll, T.
  • Journal Title: Am. J. Physiol. 290
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Attenuation of folic acid-induced renal inflammatory injury in platelet-activating factor receptor-deficient mice. (2006)
  • Author(s): Doi, K., Okamoto, K., Negishi, K., Suzuki, Y., Nakao, A., Fujita, T., Toda, A., Yokomizo, T., Kita, Y., Kihara, Y., Ishii, S., Shimizu, T., Noiri, E.
  • Journal Title: Am. J. Pathol. (印刷中)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Involvement of the platelet activating factor receptor in host defense against Streptococcus pneumoniae during postinfluenza pneumonia. (2006)
  • Author(s): van der Sluijs, K.F., van Elden, L.J.R., Nijhuis, M., Schuurman, R., Florquin, S., Shimizu, T., Ishii, S., Jansen, H.M., Lutter, R., van der Poll, T.
  • Journal Title: Am.J.Physiol. 290
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Attenuation of folic acid-induced renal inflammatory injury in platelet-activating factor receptor-deficient mice. (2006)
  • Author(s): Doi, K., Okamoto, K., Negishi, K., Suzuki, Y., Nakao, A., Fujita, T., Toda, A., Yokomizo, T., Kita, Y., Kihara, Y., Ishii, S., Shimizu, T., Noiri, E.
  • Journal Title: Am.J.Pathol. (印刷中)
• [Journal Article] Identification of T cell death-associated gene 8 (TDAG8) as a novel acid sensing G-protein-coupled receptor. (2005)
  • Author(s): Ishii, S., Kihara, Y., Shimizu, T.
  • Journal Title: J. Biol. Chem. 280 Pages: 9083-9087
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Stimulation of erythrocyte ceramide formation by platelet activating factor. (2005)
  • Author(s): Lang, P.A., Kempe, D.S., Tanneur, V., Eisele, K., Klarl, B.A., Myssina, S., Jendrossek, V., Ishii, S., Shimizu, T., Waidmann, M., Hessler, G., Huber, S.M., Lang, F., Wieder, T.
  • Journal Title: J. Cell Sci. 118 Pages: 1233-1243
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Priming effect of lipopolysaccharide on acetyl-CoA : lyso-PAF acetyltransferase is MyD88 and TRIF-independent. (2005)
  • Author(s): Shindou, H., Ishii, S., Yamamoto, M., Takeda, K., Akira, S., Shimizu, T.
  • Journal Title: J. Immunol. 175 Pages: 1177-1183
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Staphylococcal lipoteichoic acid inhibits delayed-type hypersensitivity reactions via the platelet-activating factor receptor. (2005)
  • Author(s): Zhang, Q., Mousdicas, N., Yi, Q., Al-Hassani, M., Billings, S.D., Perkins, S.M., Howard, K.M., Ishii, S., Shimizu, T., Travers, J.B.
  • Journal Title: J. Clin. Invest. 115 Pages: 2855-2861
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Recruitment of katanin p60 by phosphorylated NDEL1, a LIS1 interacting protein, is essential for mitotic cell division and neuronal migration. (2005)
  • Author(s): Toyo-oka, K., Sasaki, S., Yano, Y., Mori, D., Kobayashi, T., Toyoshima, Y.Y., Tokuoka, S.M., Ishii, S., Shimizu, T., Muramatsu, M., Hiraiwa, N., Yoshiki, A., Wynshaw-Boris, A., Hirotsune, S.
  • Journal Title: Hum. Mol. Genet. 14 Pages: 3113-3128
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Dual phase regulation of experimental allergic encephalomyelitis by platelet-activating factor. (2005)
  • Author(s): Kihara, Y., Ishii, S., Kita, Y., Toda, A., Shimada, A., Shimizu, T.
  • Journal Title: J. Exp. Med. 202 Pages: 853-863
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of T cell death-associated gene 8 (TDAG8) as a novel acid sensing G-protein-coupled receptor. (2005)
  • Author(s): Ishii.S., Kihara, Y., Shimizu, T.
  • Journal Title: J.Biol.Chem. 280 Pages: 9083-9087
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Stimulation of erythrocyte ceramide formation by platelet activating factor. (2005)
  • Author(s): Lang, P.A., Kempe, D.S., Tanneur, V., Eisele, K., Klarl, B.A., Myssina, S., Jendrossek, V., Ishii.S., Shimizu, T., Waidmann, M., Hessler, G., Huber, S.M., Lang, F., Wieder, T.
  • Journal Title: J.Cell Sci. 118 Pages: 1233-1243
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Priming effect of lipopolysaccharide on acetyl-CoA:lyso-PAF acetyltransferase is MyD88 and TRIF-independent. (2005)
  • Author(s): Shindou, H., Ishii, S., Yamamoto, M., Takeda, K., Akira, S., Shimizu, T.
  • Journal Title: J.Immunol. 175 Pages: 1177-1183
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Staphylococcal lipoteichoic acid inhibits delayed-type hypersensitivity reactions via the platelet-activating factor receptor. (2005)
  • Author(s): Zhang, Q., Mousdicas, N., Yi, Q., Al-Hassani, M., Billings, S.D., Perkins, S.M., Howard, K.M., Ishii, S., Shimizu, T., Travers, J.B.
  • Journal Title: J.Clin.Invest. 115 Pages: 2855-2861
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Recruitment of katanin p60 by phosphorylated NDEL1, a LIS1 interacting protein, is essential for mitotic cell division and neuronal migration. (2005)
  • Author(s): Toyo-oka, K., Sasaki, S., Yano, Y., Mori, D., Kobayashi, T., Toyoshima, Y.Y., Tokuoka, S.M., Ishii.S., Shimizu, T., Muramatsu, M., Hiraiwa, N., Yoshiki, A., Wynshaw-Boris, A., Hirotsune, S.
  • Journal Title: Hum.Mol.Genet. 14 Pages: 3113-3128
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dual phase regulation of experimental allergic encephalomyelitis by platelet-activating factor. (2005)
  • Author(s): Kihara, Y., Ishii, S., Kita, Y., Toda, A., Shimada, A., Shimizu, T.
  • Journal Title: J.Exp.Med. 202 Pages: 853-863
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Priming effect of lipopolysaccharide on acetyl-CoA : lyso-PAF acetyltransferase is MyD88 and TRIF-independent. (2005)
  • Author(s): Shindou, H., Ishii.S., Yamamoto, M., Takeda, K., Akira, S., Shimizu, T.
  • Journal Title: J.Immunol. 175 Pages: 1177-1183
• [Journal Article] Recruitment of katanin p60 by phosphorylated NDEL1, a LIS1 interacting protein, is essential for mitotic cell division and neuronal migration. (2005)
  • Author(s): Toyo-oka, K., Sasaki, S., Yano, Y., Mori, D., Kobayashi, T., Toyoshima, Y.Y., Tokuoka, S.M., Ishii, S., Shimizu, T., Muramatsu, M., Hiraiwa, N., Yoshiki, A., Wynshaw-Boris, A., Hirotsune, S.
  • Journal Title: Hum.Mol.Genet. 14 Pages: 3113-3128
• [Journal Article] Identification of T cell death-associated gene 8 (TDAG8) as a novel acid sensing G-protein coupled receptor. (2005)
  • Author(s): Ishii, S., Kihara, Y., Shimizu, T.
  • Journal Title: J.Biol.Chem. 280 Pages: 9083-9087
• [Journal Article] Stimulation of erythrocyte ceramide formation by platelet activating factor. (2005)
  • Author(s): Lang, P.A., Kempe, D.S., Tanneur, V., Eisele, K., Klarl, B.A., Myssina, S., Jendrossek, V., Ishii, S., Shimizu, T., Waidmann, M., Hessler, G., Huber, S.M., Lang, F., Wieder, T.
  • Journal Title: J.Cell Sci. 118 Pages: 1233-1243
• [Journal Article] Platelet-activating factor receptor develops airway hyperresponsiveness independently of airway inflammation in a murine asthma model. (2004)
  • Author(s): Ishii, S., Nagase, T., Shindou, H., Takizawa, H., Ouchi, Y., Shimizu, T.
  • Journal Title: J. Immunol. 172 Pages: 7095-7102
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Platelet activating factor receptor deficient mice have an improved host defense against pneumococcal pneumonia. (2004)
  • Author(s): Rijneveld, A.W., Weijer, S., Florquin, S., Speelman, P., Shimizu, T., Ishii, S., van der Poll, T.
  • Journal Title: J.Infect.Dis. 189 Pages: 711-716
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Trophic signals acting via phosphatidylinositol-3 kinase are required for normal pre-implantation mouse embryo development. (2004)
  • Author(s): Lu, P., Chandrakanthan, V., Cahana, A., Ishii, S., O'Neill, C.
  • Journal Title: J. Cell Sci. 117 Pages: 1567-1576
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Absence of platelet-activating factor receptor protects mice from osteoporosis following ovariectomy. (2004)
  • Author(s): Hikiji, H., Ishii, S., Shindou, H., Takato, T., Shimizu, T.
  • Journal Title: J. Clin. Invest. 114 Pages: 85-93
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Dyslipidemia associated with atherosclerotic disease systemically alters dendritic cell mobilization. (2004)
  • Author(s): Angeli, V., Llodra, J., Rong, J.X., Satoh, K., Ishii, S., Shimizu, T., Fisher, E.A., Randolph, G.J.
  • Journal Title: Immunity 21 Pages: 561-574
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Platelet-activating factor receptor deficient mice show an unaltered clearance of Haemophilus influenzae from their respiratory tract. (2004)
  • Author(s): Branger, J., Wieland, C.W., Florquin, S., Maris, N.A., Pater, J.M., Speelman, P., Shimizu, T., Ishii, S., van der Poll, T.
  • Journal Title: Shock 22 Pages: 543-547
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Platelet-activating factor receptor develops airway hyperresponsiveness independently of airway inflammation in a murine asthma model. (2004)
  • Author(s): Ishii.S., Nagase, T., Shindou, H., Takizawa, H., Ouchi, Y., Shimizu, T.
  • Journal Title: J.Immunol. 172 Pages: 7095-7102
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Trophic signals acting via phosphatidylinositol-3 kinase are required for normal pre-implantation mouse embryo development. (2004)
  • Author(s): Lu, D.P., Chandrakanthan, V., Cabana, A., Ishii, S., O'Neill, C.
  • Journal Title: J.Cell Sci. 117 Pages: 1567-1576
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Absence of platelet-activating factor receptor protects mice from osteoporosis following ovariectomy. (2004)
  • Author(s): Hikiji, H., Ishii.S., Shindou, H., Takato, T., Shimizu, T.
  • Journal Title: J.Clin.Invest. 114 Pages: 85-93
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dyslipidemia associated with atherosclerotic disease systemically alters dendritic cell mobilization. (2004)
  • Author(s): Angeli, V, Llodra, J., Rong, J.X., Satoh, K, Ishii, S., Shimizu, T., Fisher, E.A., Randolph, G.J
  • Journal Title: Immunity 21 Pages: 561-574
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Trophic signals acting via phosphaidylinositol-3 kinase are required for normal pre-implantation mouse embryo development. (2004)
  • Author(s): Lu, P., Chandrakanthan, V., Cahana, A., Ishii, S., O'Neill, C.
  • Journal Title: J.Cell Sci. 117 Pages: 1567-1576
• [Journal Article] Platelet-activating factor receptor develops airway hyperresponsiveness independently of airway inflammation in a murine asthma model. (2004)
  • Author(s): Ishii, S., Nagase, T., Shindou, H., Takizawa, H., Ouchi, Y., Shimizu, T.
  • Journal Title: J.Immunol. 172 Pages: 7095-7102
• [Journal Article] Absence of platelet-activating factor receptor protects mice from osteoporosis following ovariectomy. (2004)
  • Author(s): Hikiji, H., Ishii, S., Shindou, H., Takato, T., Shimizu, T.
  • Journal Title: J.Clin.Invest. 114 Pages: 85-93
• [Journal Article] Attenuation of folic acid-induced renal inflammatory injury in platelet-activating factor receptor-deficient mice.
  • Author(s): Doi, K., Okamoto, K., Negishi, K., Suzuki, Y., Nakao, A., Fujita, T., Toda, A., Yokomizo, T., Kita, Y., Kihara, Y., Ishii, S., Shimizu, T., Noiri, E.
  • Journal Title: Am.J.Pathol. (in press)
  • Description: 「研究成果報告書概要(欧文)」より