A study for role of PGE2 on adhesion of mast cells to fibronectin

• Project/Area Number: 15390024
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Mukogawa Women's University
• Principal Investigator: ICHIKAWA Atsushi Mukogawa Women's University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (10025695)
• Co-Investigator(Kenkyū-buntansha): SENMA Masanori Mukogawa Women's University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (90098533) ; SUGIMOTO Yukihiko Kyoto University, Associate Professor, 薬学研究科, 助教授 (80243038) ; TANAKA Satoshi Kyoto University, Lecturer, 薬学研究科, 助手 (40303846)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥15,300,000 (Direct Cost: ¥15,300,000); Fiscal Year 2004: ¥7,400,000 (Direct Cost: ¥7,400,000); Fiscal Year 2003: ¥7,900,000 (Direct Cost: ¥7,900,000)
• Keywords: mast cells / cell adhesion / prostaglandin E2 / Extracellular matrix / cAMP / Epac / Ca2+ / PGE2 / 細胞外マトリックス / PKA / mastocytoma P-815 / Epac-Rapシグナル / EP4サブタイプ / フィブロネクチン / Ca^<2+> / アレルギー

Research Abstract

The purpose of this project is to determine the mechanism of mast cell adhesion to fibronectin. Mastocytoma P-815 cells were cultured on a plate pre-coated with or without fibronectin in 5%-FCS containing Fisher medium, and incubated with PGE2 for various times at 37℃ in 5% C02-incubator. Obtained results are shown below ;
1.P-815 cells were adhered to fibronectin in response to 10^<-8> M-10^<-6> M PGE2. The adherence activity of PGE2 was appeared through its binding to EP4 receptors expressed on P-815 cells, followed by a stimulated cAMP formation and cAMP-dependent protein kinase A activity. These responses were not demonstrated up to 4-5 hrs after PGE2 addition, indicating involvement of newly formed unknown proteins, which have been searching thereafter.
2.After initiation of PGE2-induced P-815 cell adhesion to fibronectin, being inhibited by PKA inhibitor H-89, adhered P-815 cells did not leak out by H89 treatment. However, PGE2-adhered P-815 cells were easily detached by a decrease of increasing intracellular Ca2+ level by treatment with EGTA,BAPTA/AM, and SKF96365, a Ca2+ channel ROCC inhibitor. Really, adhered P-815 cells were found to consist of high intracellular Ca2+ level.
3.PGE2-activated P-815 cell adhesion to fibronectin were mostly suppressed by incubation with CaM kinase inhibitor.
4.CAMP/PKA-independent signals were extensively searched for, and as a result have determined involvement of Epac signal (exchange protein directly activated by cAMP) as important candidatet molecules.
From these results, we have presented that PGE2-activated P-815 cell adhesion might be comprised of two signals, first during binding initiation phase cAMP/PKA dependent reactions involved in while during late binding maintenance phase Epac reactions did. These results are surely important to resolve mast cell adhesion ability to extra-cellular matrix like ibronectin in local inflammatory tissues.

Research Products

• [Journal Article] A cluster of aromatic amino acids in the i2 loop plays a key role for Gs coupling in prostaglandin EP2 and EP3 receptors (2004)
  • Author(s): Sugimoto, S., Nakato, T., Kita, A., Tabara, H., Tanaka, S., Ichikawa, A.
  • Journal Title: J.Biol.Chem. 279 Pages: 11016-11026
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Functional domains essential for Gs activity in prostaglandin EP2 and EP3 receptors (2004)
  • Author(s): Sugimoto, S., Nakato, T., Kita, A., Hatae, S., Ichikawa, A.
  • Journal Title: Life Science 74 Pages: 135-141
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Insight into prostanoid functions : Lession from receptor-knockout mice (2004)
  • Author(s): Sugimoto, Y., Narumiya, S., Ichikawa, A.
  • Journal Title: The eicosanoids(ed by P.Curitis-Pior, John Willy, Sons) Pages: 219-225
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression of L-histidine decarboxykase in granules of elicited mouse mastocytoma P-815 cells (2004)
  • Author(s): Tanaka, S., Deai, K., Takahashi, K., Yamane, H., Sugimoto, Y., Ichikawa, A.
  • Journal Title: Eur.J.Immunol 34 Pages: 1472-1482
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A cluster of aromatic amino acids in the i2 loop plays a key role for Gs coupling in prostaglandin EP2 and EP3 receptors. (2004)
  • Author(s): Sugimoto, Y., Nakato, T., Kita, A., Takahashi, Y., Hatae, N., Tabata, H., Tanaka, S., Ichikawa, A.
  • Journal Title: J.Biol.Chem. 279 Pages: 11016-11026
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Functional domains essential for Gs activity in prostaglandin EP2 and EP3 receptors. (2004)
  • Author(s): Sugimoto, Y., Nakato, T., Kita, A., Hatae, N., Tabata, H., Tanaka, S., Ichikawa, A.
  • Journal Title: Life Sci. 74 Pages: 135-141
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Insight into prostanoid functions : Lession from receptor-knockout mice (2004)
  • Author(s): Sugimoto, Y., Narumiya, S., Ichikawa, A.
  • Journal Title: The eicosanoids (ed by Pior, John Willy & Sons) Pages: 219-225
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A cluster of aromatic amino acids in the i2 loop plays a key role for Gs coupling in prostglandin EP2 and EP3 receptors (2004)
  • Author(s): Sugimoto, Y., Nakato, T., Kita, A., Takahshi, Y., Hatae, N., Tabata, H., Tanaka, S., Ichikawa, A
  • Journal Title: J.Biol.Chem. 279 Pages: 11016-11026
• [Journal Article] Prostanoid receptor is involved in suppression of 3T3-L1 adipocyte differentiation (2004)
  • Author(s): Tsuboi, H., Sugimoto, Y., Kainoh, T., Ichikawa, A
  • Journal Title: Biochem.Biophys.Res.Commun. 322 Pages: 1066-1072
• [Journal Article] Microarray evaluation of EP4 receptor-mediated prostagalandin E(2) suppression of 3T3-L1 adipocyte differentiation (2004)
  • Author(s): Sugimoto, Y., Tsuboi, H., Okuno, Y., Tamba, S, Tsuchiya, S., Tsujimoto, C., Ichikawa, A.
  • Journal Title: Biochem.Biophys.Res, Commun. 322 Pages: 911-917
• [Journal Article] Role of cellular Ca2+ for PGE2-stimulated activity of mastocytoma P-815 cells for fibronectin (2004)
  • Author(s): Sakanaka, M., Hatae, N., Tanaka, S., Senma, M., Ichikawa, A.
  • Journal Title: Seikagaku 76 Pages: 1071-1071
• [Journal Article] Induction of adherent activity in mastocytoma P-815 cells by the cooperation of two prostaglandin E2 receptor subtypes, EP3 and EP4 (2003)
  • Author(s): Hatae, N., Kita, A., Sugimoto, Y., Tanaka, S., Ichikawa, A.
  • Journal Title: J.Biol.Chem. 278 Pages: 17977-17981
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characteristics of thermoregulatory and febrile responses in mice deficient in prostaglandin EP1 and EP3 receptors (2003)
  • Author(s): Oka, T., Oka, K., Kobayashi, T., Ichikawa, A., Ushikubi, F., Narumiya, S.
  • Journal Title: J.Physiol. 551 Pages: 945-954
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction of adherent activity in nastocytoma P-815 cells by the cooperation of two prostaglandin E2 receptor subtypes, EP3 and EP4. (2003)
  • Author(s): Hatae, N., Kita, A., Tanaka, A., Sugimoto, Y., Ichikawa, A.
  • Journal Title: J.Biol.Chem. 278 Pages: 17977-17981
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characteristics of thermoregulatory and febrile responses in mice deficient in prostaglandin EP1 and EP3 receptors. (2003)
  • Author(s): Oka, T., Oka, K., Kobayashi, T., Sugimoto, Y., Ichikawa, A., Ushikubi, F., Narumiya, S., Saper, C.B.
  • Journal Title: J.Physiol. 551 Pages: 945-954
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification and characterization of a novel progesterone receptor-binding element in the mouse prostaglandin E receptor subtype EP2 gene. (2003)
  • Author(s): Tsuchiya, S., Tanaka, S., Sugimoto, Y., Katsuyama, M., Ikegami, R., Ichikawa, A.
  • Journal Title: Genes to Cells 8 Pages: 747-758
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction of adherent activity in mastocytoma P-815 cells by the cooperation of two prostaglandin E2 receptor subtypes EP3 and EP4 (2003)
  • Author(s): Hatae, N., Kita, A., tanaka, S., Sugimoto, Y., Ichikawa, A.
  • Journal Title: J.Biol.Chem. 278 Pages: 17977-17981
• [Journal Article] Identification and characterization of a novel progesterone receptor-binding element in the mouse prostagiandin E receptor subtypes EP2 gene (2003)
  • Author(s): Tsuchiya, S., Takana, S.Sugimoto, Y., Katsuyama, M., Ikegami, R., Ichikawa, A.
  • Journal Title: Genes to Cells 8 Pages: 747-758
• [Publications] Sugimoto, Y., et al.: "A cluster of aromatic amino acids in the i2 loop plays a key role for Gs coupling in prostaglandin EP2 and EP3 receptors."J.Biol.Chem. 279. 11016-11026 (2004)
• [Publications] Sugimoto, Y., et al.: "Functional domains essential for Gs activity in prostaglandin EP2 and EP3 receptors."Life Sci.. 74. 135-141 (2003)
• [Publications] Tsuchiya, S., et al.: "Identification and characterization of a novel progesterone receptor-binding element in the mouse prostaglandin E receptor subtype EP2 gene."Genes to Cells. 8. 747-758 (2003)
• [Publications] Oka, T., et al.: "Characteristics of thermoregulatory and febrile responses in mice deficient in prostaglandin EP1 and EP3 receptors."J.Physiol.. 551. 945-954 (2003)
• [Publications] Hatae, N., et al.: "Induction of adherent activity in mastocytoma P-815 cells by the Cooperation of two prostaglandin E2 receptor subtypes, EP3 and EP4."J.Biol.Chem. 278. 17977-17981 (2003)
• [Publications] Amano, H., et al.: "Host prostaglandin E2-EP3 signaling regulates tumor-associated angiogenesis and tumor growth."J.Exp.Med.. 197. 221-232 (2003)