Characterization of functional P2X1 receptors in mouse megakaryocytes
Abstract Introduction Although accumulating evidence within the past 5 years strongly supports the importance of platelet P2X1 receptors in hemostasis and thrombosis, P2X1 receptors of platelet and/or its progenitor cell, megakaryocyte, have not been fully characterized. The aim of this study was to...
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| Published in | Thrombosis Research Vol. 119; no. 3; pp. 343 - 353 |
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| Main Author | |
| Format | Journal Article |
| Language | English |
| Published |
New York, NY
Elsevier BV
2007
Elsevier Science |
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| Online Access | Get full text |
| ISSN | 0049-3848 |
| DOI | 10.1016/j.thromres.2006.03.007 |
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| Abstract | Abstract Introduction Although accumulating evidence within the past 5 years strongly supports the importance of platelet P2X1 receptors in hemostasis and thrombosis, P2X1 receptors of platelet and/or its progenitor cell, megakaryocyte, have not been fully characterized. The aim of this study was to electrophysiologically and pharmacologically characterize the functional P2X1 receptors on mouse megakaryocytes. Materials and methods The currents in response to nucleotides were examined using the patch-clamp whole-cell recording. Results The agonist profile of megakaryocyte P2X1 receptors was ATP > α,β-methylene ATP > β,γ-methylene ATP. The P2X1 receptors exhibited substantial monovalent as well as divalent cation permeability and the ratios of Na+ to Cs+ and Ca2+ to Cs+ permeability were 1 and 2.5, respectively. P2X receptor antagonists except suramin significantly inhibited the P2X1 responses with an IC50 values of 0.4 μM for pyridoxal-phosphate-6-azophenyl-2′,4′-disulfonate (PPADS), 0.3 μM for 2′,3′- O -(2,4,6-trinitophenyl)-adenosine 5′-triphosphate (TNP-ATP), 20 μM for reactive blue 2 (RB2), or 160 μM for 8,8′-(carbonylbis(imino-3,1-phenylene carbonylimino)bis(1,3,5-naphthalenetrisulfonic acid) (NF023), respectively. Suramin had no significant effect on the P2X1 responses. In rat megakaryocytes, suramin similarly had no significant effect on the P2X1 responses, but abolished the P2Y receptor-mediated responses, indicating that the suramin was active under present experimental condition. Conclusions These results provide the basic properties of mouse megakaryocyte P2X1 receptors and would be helpful to examine the P2 receptor signaling in platelets and megakaryocytes. |
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| AbstractList | Abstract Introduction Although accumulating evidence within the past 5 years strongly supports the importance of platelet P2X1 receptors in hemostasis and thrombosis, P2X1 receptors of platelet and/or its progenitor cell, megakaryocyte, have not been fully characterized. The aim of this study was to electrophysiologically and pharmacologically characterize the functional P2X1 receptors on mouse megakaryocytes. Materials and methods The currents in response to nucleotides were examined using the patch-clamp whole-cell recording. Results The agonist profile of megakaryocyte P2X1 receptors was ATP > α,β-methylene ATP > β,γ-methylene ATP. The P2X1 receptors exhibited substantial monovalent as well as divalent cation permeability and the ratios of Na+ to Cs+ and Ca2+ to Cs+ permeability were 1 and 2.5, respectively. P2X receptor antagonists except suramin significantly inhibited the P2X1 responses with an IC50 values of 0.4 μM for pyridoxal-phosphate-6-azophenyl-2′,4′-disulfonate (PPADS), 0.3 μM for 2′,3′- O -(2,4,6-trinitophenyl)-adenosine 5′-triphosphate (TNP-ATP), 20 μM for reactive blue 2 (RB2), or 160 μM for 8,8′-(carbonylbis(imino-3,1-phenylene carbonylimino)bis(1,3,5-naphthalenetrisulfonic acid) (NF023), respectively. Suramin had no significant effect on the P2X1 responses. In rat megakaryocytes, suramin similarly had no significant effect on the P2X1 responses, but abolished the P2Y receptor-mediated responses, indicating that the suramin was active under present experimental condition. Conclusions These results provide the basic properties of mouse megakaryocyte P2X1 receptors and would be helpful to examine the P2 receptor signaling in platelets and megakaryocytes. |
| Author | Ikeda, Masahiro |
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| Keywords | RB2 8,8′-(carbonylbis(imino-3,1-phenylene carbonylimino)bis(1,3,5-naphthalenetrisulfonic acid) Patch-clamp αβ-meATP ethylene glycol bis(β-aminoethylether)- N, N, N′, N′-tetraacetic acid NF023 pyridoxal-phosphate-6-azophenyl-2′,4′-disulfonic acid Megakaryocytes PPADS β,γ-methylene ATP βγ-meATP TNP-ATP α,β-methylene ATP 2′,3′- O-(2,4,6-trinitrophenyl)-adenosine 5′-triphosphate reactive blue 2 IS ES internal solution S.E.M EGTA external solution P2X 1 standard error of the mean Platelets ATP Purine nucleoside Rat Megakaryocyte Cardiovascular disease Suramin sodium Vascular disease Antiprotozoal agent Hemostasis Antiviral P2X1 Adenosine Blue Rodentia Triphosphates Permeability Thrombosis Vertebrata Platelet Mammalia Clamping(surgery) Mouse Animal Parasiticide Patch Progenitor cell |
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| SubjectTerms | Adenosine Triphosphate Animals Antineoplastic Agents Biological and medical sciences Blood and lymphatic vessels Blood coagulation Blood vessels and receptors Cardiology. Vascular system Cations, Divalent Cations, Monovalent Cell Membrane Permeability Diseases of the peripheral vessels. Diseases of the vena cava. Miscellaneous Fundamental and applied biological sciences. Psychology Hematology, Oncology and Palliative Medicine Investigative techniques, diagnostic techniques (general aspects) Ion Transport Medical sciences Megakaryocytes Membrane Potentials Mice Purinergic P2 Receptor Agonists Purinergic P2 Receptor Antagonists Rats Receptors, Purinergic P2 Receptors, Purinergic P2X Suramin Vertebrates: cardiovascular system |
| Title | Characterization of functional P2X1 receptors in mouse megakaryocytes |
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