| Project/Area Number |
13558114
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Nagoya City University |
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Principal Investigator |
UGAWA Shinya Nagoya City University, Graduate School of Medical Sciences, Research Associate., 大学院・医学研究科, 助手 (20326135)
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| Co-Investigator(Kenkyū-buntansha) |
HIRABAYASHI Yoshifumi Nagoya City University, Graduate School of Medical Sciences, Assistant Professor., 大学院・医学研究科, 講師 (30181184)
FUJIMORI Osamu Nagoya City University, Graduate School of Medical Sciences, Associate Professor., 大学院・医学研究科, 助教授 (30128350)
SHIMADA Shoichi Nagoya City University, Graduate School of Medical Sciences, Professor., 大学院・医学研究科, 教授 (20216063)
UEDA Takashi Nagoya City University, Graduate School of Medical Sciences, Research Associate., 大学院・医学研究科, 助手 (90244540)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2003: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2002: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2001: ¥7,300,000 (Direct Cost: ¥7,300,000)
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| Keywords | Sour receptor / Taste sensation / Cloning / MDEG / Adaptation / Gene / Heteromer / ASIC / 味覚受容体 / アミロライド感受性陽イオンチャンネル / T2Rfamily / Oocyte Electrophysiology / カルシウムイメージング / 味覚修飾物質 |
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| Research Abstract |
Acid-sensing ion channel-2a (ASIC2a) is an amiloride-blockable proton-gated action channel, probably contribution to sour-taste detection in rat taste cells. To isolate another subtype of the sour-taste receptor, we screened a rat circumvallate papilla cDNA library and identified ASIC2b, an N-terminal splice variant of ASICA2a. Reverse transcription-polymerase chain reaction analyses confirmed the expression of ASIC2b transcripts in the circumvallate papilla and, moreover, demonstrated its expression in the foliate and fungiform papillae. Immunohistochemical analyses revealed that ASIC2b, as well as ASIC2a, was expressed in a subpopulation of taste cells in the circumvallate, foliate, and fungiform papillae, and some of the cells displayed both ASIC2a-and ASIC2b-immunoreactivities. Subsequent co-immunoprecipitation studies with circumvallate papillae extracts indicated that ASIC2b associated with ASIC2a to form assemblies, and, together with our immunohistochemical findings, strongly suggested that both ASIC2 subunits formed heteromeric channels in taste cells in the circumvallate, foliate, and fungiform papillae. Oocyte electro-physiology demonstrated that the ASIC2a/ASIC2b channel generated maximal inward currents at pH 2.0, which is in good agreement with the in vivo pH-sensitivity of rat taste cells, and that the amiloride-sensitivity of the heteromer decreased with decreasing pH and was almost completely abolished at pH 2.0. These findings provide persuasive explanations for the amiloride-insensitivity of acid-induced responses of rat taste cells.
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