1997 年度実績報告書

脂質平面膜を用いた膜電位感受性色素の評価系の開発とその応用

研究課題

研究課題/領域番号	07557143
研究機関	東京大学
研究代表者	桐野豊東京大学, 大学院・薬学系研究科, 教授 (10012668)
研究分担者	浜岡勤富士写真フィルム(株), 機器事業本部, 部長(研究職) 平嶋尚英東京大学, 大学院・薬学系研究科, 助手 (10192296) 川原茂敬東京大学, 大学院・薬学系研究科, 助手 (10204752)
キーワード	膜電位感受性蛍光色素 / 異質平面膜 / 冷却CCDカメラ / 高速差分増幅式カメラ / ナメクジ中枢神経 / 嗅覚-味覚連合学習
研究概要	1.システムの改良:これまで、平面脂質二重膜に結合した膜電位感受性色素の蛍光をシングルホトダイオードカメラで測定していたが、これをノイズが小さく、受講面が大きくて高感度な冷却CCDカメラ(アストロカム社、KAF1400)に換えたところ、明確に、蛍光強度の膜電位依存性が測定できるようになった。従来より、神経節等の膜電位測定に経験的によく用いられてきた色素であるDi-4-ANEPPSは、膜電位に対する蛍光強度変化が大きいものであることが確認できた。 2.生体膜への応用:感覚器(嗅覚器としての大小触覚、あるいは、味覚器としての唇)を付けて切り出したナメクジの脳を膜電位感受性蛍光色素Di-4-ANEPPSで染色し、膜電位の時間的空間的パターンの変化を高速差分増幅式カメラ(富士写真フィルム、Deltaron 1700)により測定した。大触角に匂いを与えると、匂いの種類によらず、脳神経節前脳部の膜電位振動の周波数が増大した。小触覚に匂いを与えると、ナメクジが好む食物の匂いの場合には振動数が増大し、ナメクジが忌避する物質の匂いの場合には振動数が減少した。また、ナメクジが忌避する味物質を唇に適用したときには、前脳の膜電位が強く過分極方向にシフトした。更に、小触角に嗅覚-味覚連合学習により忌避性となった食物の匂いを与えた後の数秒間、特異的な興奮性を示す一群の細胞が存在することが見出された。このように、本研究で開発された膜電位感受性蛍光色素を用いる膜電位の光学的測定法は、脳における味覚・嗅覚情報処理機構や学習・記憶の機構に極めて有用な実験手法となることを示すことが出来た。

研究成果

(22件)

すべてその他

すべて文献書誌 (22件)

[文献書誌] Atsuhisa Tomita: "Removal after Addition of NO-generating Agents and 8-Bromo-cyclic GMP Cause Morphological Change of Cultured Cerebellar Astrocytes : a New Mode of NO Action" Brain Res.744. 344-346 (1997)
[文献書誌] Toshiharu Suzuki: "Phosphorylation of Alzheimer's β-Amyloid Precursor-like Proteins" Biochemistry. 36. 4643-4649 (1997)
[文献書誌] Hiroshi Tokumaru: "Involvement of Calciseptine-sensitive Calcium Channels in Acetylcholine Release from Electric Organ Synaptosomes" Biol.Pharma.Bull.20. 641-645 (1997)
[文献書誌] Shigenori Kawahara: "Comparative Study on Neural Oscillation in the Procerebrum of the Terrestrial Slugs Incilaria bilineata and Limax marginatus" J.Ex.Biol.200. 1851-1861 (1997)
[文献書誌] J.de.Barry: "Time-resolved Imaging of Protein Kinase C Activation during Sea Urchin Egg Fertilization" Exp.Cell Res.234. 115-124 (1997)
[文献書誌] Katsumi Matsuzaki: "Membrane Permeabilization Mechanisms of a Cyclic Antimicrobial Peptide,Tachyplesin I,and Its Linear Analog" Biochemistry. 36. 9799-9806 (1997)
[文献書誌] Toru Ishizuka: "Molecular Cloning of Narke japonica Electric Organ Synaphins/Complexins Involved in Transmitter Release" Neurosci.Lett.232. 107-110 (1997)
[文献書誌] Yosushi Kishimoto: "Conditioned Eyeblink Response Is Impaired in Mutant Mice Lacking NMDA Receptor Subunit NR2A" NeuroReport. 8. 3717-3721 (1997)
[文献書誌] Hirotatsu Kojima: "Development of a Fluorescent Indicator for the Bioimaging of Nitric Oxide" Biol.Pharm.Bull.20. 1229-1232 (1997)
[文献書誌] 川原茂敬: "神経細胞のカルシウムイメージング" 細胞. 30. 7-10 (1997)
[文献書誌] Yong I.Kim: "An Autoimmune Animal Model of the Lambert-Eaton Syndrom" Ann.N.Y.Acad.Sci. (in press). (1998)
[文献書誌] Yasushi Satoh: "Activation of Adenosine A1 and A2 Receptors Differentially Affects Acetylcholine Release from Electric Organ Synaptosomes through Modulation of Calcium Channels" Neurosci.Res.(in press). (1998)
[文献書誌] Ko-ichi Iijima: "Phosphorylated Form of Alzheimer's Amyloid Precursor Protein Exists in Cholinergic Nerve Termini from Electric Ray Electric Organ" Biochem.J.(in press). (1998)
[文献書誌] Satoshi Watanabe: "Morphological Characterization of the Bursting and Nonbursting Neurons in the Olfactory Center of the Terrestrial Slug Limax marginatus" J.Exp.Biol.(in press). (1998)
[文献書誌] Yasushi Satoh: "Lambert-Eaton Syndrome Antibodies Inhibit Acetylcholine Release and P/Q-type Ca^<2+> Channels in Electric Ray Nerve Endings" J.Physiol.(in press). (1998)
[文献書誌] Susumu Tomita: "Cleavage of Alzheimer's Amyloid Precursor Protein by Secretases Occurs after O-glycosylation of APP in Protein Secretory Pathway" J.Biol.Chem.(in press). (1998)
[文献書誌] Shigeo Watanabe: "Differential Roles of Two Types of Voltage-gated Ca^<2+> Channels in the Dendrites of Rat Cerebellar Purkinje Neurons" Brain Res.(in press). (1998)
[文献書誌] Tetsuya Kimura: "Behavioral Modulation Induced by Food-Odor Aversive Conditioning,and Its Influence on the Olfactory Responses of an Oscillatory Brain Network in the Slug,Limax marginatus" Learning and Memory. (in press). (1998)
[文献書誌] Tetsuya Kimura: "Optical Recording Analysis of Olfactory Response of the PC Lobe in the Slug Brain" Leaming and Memory. (in press). (1998)
[文献書誌] Yutaka Kirino: "The Role of Adenosine in the Nervous System" Elsevier, 352 (1997)
[文献書誌] 桐野豊: "カルシウムシグナリング" 培風館, 173 (1997)
[文献書誌] 桐野豊: "基礎薬学物理化学" 廣川書店, 342 (1997)

1997 年度 実績報告書

脂質平面膜を用いた膜電位感受性色素の評価系の開発とその応用

研究代表者

桐野 豊 東京大学, 大学院・薬学系研究科, 教授 (10012668)

研究成果

[文献書誌] Atsuhisa Tomita: "Removal after Addition of NO-generating Agents and 8-Bromo-cyclic GMP Cause Morphological Change of Cultured Cerebellar Astrocytes : a New Mode of NO Action" Brain Res.744. 344-346 (1997)

[文献書誌] Toshiharu Suzuki: "Phosphorylation of Alzheimer's β-Amyloid Precursor-like Proteins" Biochemistry. 36. 4643-4649 (1997)

[文献書誌] Hiroshi Tokumaru: "Involvement of Calciseptine-sensitive Calcium Channels in Acetylcholine Release from Electric Organ Synaptosomes" Biol.Pharma.Bull.20. 641-645 (1997)

[文献書誌] Shigenori Kawahara: "Comparative Study on Neural Oscillation in the Procerebrum of the Terrestrial Slugs Incilaria bilineata and Limax marginatus" J.Ex.Biol.200. 1851-1861 (1997)

[文献書誌] J.de.Barry: "Time-resolved Imaging of Protein Kinase C Activation during Sea Urchin Egg Fertilization" Exp.Cell Res.234. 115-124 (1997)

[文献書誌] Katsumi Matsuzaki: "Membrane Permeabilization Mechanisms of a Cyclic Antimicrobial Peptide,Tachyplesin I,and Its Linear Analog" Biochemistry. 36. 9799-9806 (1997)

[文献書誌] Toru Ishizuka: "Molecular Cloning of Narke japonica Electric Organ Synaphins/Complexins Involved in Transmitter Release" Neurosci.Lett.232. 107-110 (1997)

[文献書誌] Yosushi Kishimoto: "Conditioned Eyeblink Response Is Impaired in Mutant Mice Lacking NMDA Receptor Subunit NR2A" NeuroReport. 8. 3717-3721 (1997)

[文献書誌] Hirotatsu Kojima: "Development of a Fluorescent Indicator for the Bioimaging of Nitric Oxide" Biol.Pharm.Bull.20. 1229-1232 (1997)

[文献書誌] 川原茂敬: "神経細胞のカルシウムイメージング" 細胞. 30. 7-10 (1997)

[文献書誌] Yong I.Kim: "An Autoimmune Animal Model of the Lambert-Eaton Syndrom" Ann.N.Y.Acad.Sci. (in press). (1998)

[文献書誌] Yasushi Satoh: "Activation of Adenosine A1 and A2 Receptors Differentially Affects Acetylcholine Release from Electric Organ Synaptosomes through Modulation of Calcium Channels" Neurosci.Res.(in press). (1998)

[文献書誌] Ko-ichi Iijima: "Phosphorylated Form of Alzheimer's Amyloid Precursor Protein Exists in Cholinergic Nerve Termini from Electric Ray Electric Organ" Biochem.J.(in press). (1998)

[文献書誌] Satoshi Watanabe: "Morphological Characterization of the Bursting and Nonbursting Neurons in the Olfactory Center of the Terrestrial Slug Limax marginatus" J.Exp.Biol.(in press). (1998)

[文献書誌] Yasushi Satoh: "Lambert-Eaton Syndrome Antibodies Inhibit Acetylcholine Release and P/Q-type Ca^<2+> Channels in Electric Ray Nerve Endings" J.Physiol.(in press). (1998)

[文献書誌] Susumu Tomita: "Cleavage of Alzheimer's Amyloid Precursor Protein by Secretases Occurs after O-glycosylation of APP in Protein Secretory Pathway" J.Biol.Chem.(in press). (1998)

[文献書誌] Shigeo Watanabe: "Differential Roles of Two Types of Voltage-gated Ca^<2+> Channels in the Dendrites of Rat Cerebellar Purkinje Neurons" Brain Res.(in press). (1998)

[文献書誌] Tetsuya Kimura: "Behavioral Modulation Induced by Food-Odor Aversive Conditioning,and Its Influence on the Olfactory Responses of an Oscillatory Brain Network in the Slug,Limax marginatus" Learning and Memory. (in press). (1998)

[文献書誌] Tetsuya Kimura: "Optical Recording Analysis of Olfactory Response of the PC Lobe in the Slug Brain" Leaming and Memory. (in press). (1998)

[文献書誌] Yutaka Kirino: "The Role of Adenosine in the Nervous System" Elsevier, 352 (1997)

[文献書誌] 桐野 豊: "カルシウムシグナリング" 培風館, 173 (1997)

[文献書誌] 桐野 豊: "基礎薬学 物理化学" 廣川書店, 342 (1997)

1997 年度実績報告書

桐野豊東京大学, 大学院・薬学系研究科, 教授 (10012668)

[文献書誌] 桐野豊: "カルシウムシグナリング" 培風館, 173 (1997)

[文献書誌] 桐野豊: "基礎薬学物理化学" 廣川書店, 342 (1997)