1998 Fiscal Year Annual Research Report

モルモット単離小脳を用いたin vitro学習系の開発

Research Project

Project/Area Number	10877356
Research Institution	The University of Tokyo
Principal Investigator	桐野豊東京大学, 大学院・薬学系研究科, 教授 (10012668)
Co-Investigator(Kenkyū-buntansha)	川原茂敬東京大学, 大学院・薬学系研究科, 助手 (10204752)
Keywords	瞬目反射 / 古典的条件付け / 学習 / モルモット / 小脳 / 単離脳
Research Abstract	本研究は、学習記憶の神経回路メカニズムを明らかにする研究の一環としてin vitro学習が可能な単離脳標本を開発することを目標としている。 1. 個体レベルにおける学習パラダイムの検討: 瞬目反射条件付けは音または光を条件刺激(CS)とし,まぶたへの侵害刺激を無条件刺激(US)とする古典的条件付けの一つである。この学習には小脳が必須であることがウサギを用いた研究により明らかになっている。in situおよびin vitro脳標本における条件づけを念頭に、モルモットにおける学習条件を検討した。まず、典型的な遅延パラダイムで連続500回(約4時間)の条件付けを行ったところ、学習可能であることが明らかとなった。次に、CSを1、5、および10kHzにして通常の条件付け(100回×6日間)を行ったところ、どの周波数においても学習は成立するが、高い周波数の方が速く学習する傾向があった。また、CSとして音の代わりに光を用いた場合にも学習が可能であることも明らかとなった。CSとUSの時間間隔に関しては、250msおよび500msの遅延パラダイムで学習可能であり、また、痕跡パラダイムでも100msおよび500msで学習可能であることが明らかとなった。 2. in vitro脳標本作成の条件検討: モルモットより聴神経(第8神経。条件刺激の入力経路),三叉神経(無条件刺激の入力経路),および外転神経(応答の出力経路)をつけた小脳一脳幹血管潅流標本を作成した。ついで、USに対する無条件応答(UR)がこの標本で維持されていることを確認するために、外転神経より細胞外記録をとりながら三叉神経を電気刺激したところ、いくつかの標本においては記録する事ができた。来年度は、標本作成法や記録方法を改善し、URが維持されている標本において第8神経と三叉神経の同時刺激で条件付けが可能かどうか検討する予定である。

Research Products
(20 results)

All Other

All Publications (20 results)

[Publications] Ko-ichi Iijima: "Phosphorylated Form of Alzheimer's Amyloid Precursor Protein Exists in Cholinergic Nerve T ermini from Electric Ray Electric Organ" Biochem.J.330. 29-33 (1998)
[Publications] Susumu Tomita: "Cleavage of Alzheimer's Amyloid Precursor Protein by Secretases Occure after O-glycosyla-tion of APP in Protein Secretory Pathway" J.Biol.Chem.273. 6277-6284 (1998)
[Publications] Hirotasu Kojima: "Development of a Fluorescent Indicator for Nitric Oxide Based on the Fluorescein Chromophore" Chem. Pharm. bull.46. 373-375 (1998)
[Publications] Yasushi Satoh: "Lambert-Eaton Syndrome Antibodies Inhibit Acetylcholine Release and P/Q-type Ca^<2+> Channels in Electric Ray Nerve Endings" J.Physiol.508. 427-438 (1998)
[Publications] Tetsuya Kimura: "Behavioral Modulation Indced 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. 4. 365-375 (1998)
[Publications] Tetsuya Kimura: "Optical Recording Analysis of Olfactory Response of the PC Lobe in the Slug Brain" Learning and Memory. 4. 389-400 (1998)
[Publications] Satoshi Watanabe: "Morphological Characterization of the Bursting and Nonbursting Neurons in the Olfactory Center of the Terrestrial Slug Limax marginatus" J.Exp.Biol.201. 925-930 (1998)
[Publications] Yong I.Kim: "An Autoimmune Animal Model of the Lambert-Eaton Syndrom" Ann.N.Y.Acad.Sci. 841. 670-676 (1998)
[Publications] Shigeo Watanabe: "Differential Roles of Two Types of Voltage-gated Ca^<2+> Channels in the Dendrites of Rat Cerebellar Purkinje Neurons" Brain Res.791. 43-55 (1998)
[Publications] Susumu Tomita: "A Basic Amino Acid in the Cytopolasmic Domain of Altzheimer's β-Amyloid Precursor Protein(APP)Is Essential for Cleavage of APP at the α-site" J.Biol.Chem.273. 19304-19310 (1998)
[Publications] Hirotatsu Kojima: "Detection and Imaging of Nitric Oxide with Novel Fluorescent Indicators:Diaminofluoresceins" Anal.Chem.70. 2446-2453 (1998)
[Publications] 桐野豊: "学習記憶研究の戦略" 放射線医学. 38. 174-177 (1998)
[Publications] Dong-Seok Lee: "Phospholipid Translocation from Outer to Inner Leafled of Synaptic Vescle membranes Isolated from the Electric Organ of Japanese Electric Ray Narke japonica" J.Biochem.124. 798-803 (1998)
[Publications] Tsuyoshi Inoue: "Selective Optical Recording of the Neural Activity in the Olfactory Center of Land Slug Using a Calcium Indicator Dye" Bioimages. 6. 59-67 (1998)
[Publications] Azusa Nakagawa-Inoue: "Odorant-evoked Increase Cytosolic Free Calcium in Cultured Anntenal Neurons of Blowflies" Zool.Sci.15. 661-666 (1998)
[Publications] Yutaka Hirakura: "Membrane Perturbation by the Alzhemer Amyloid Fragment β25-35 Requires Aggregation" Biochem.Mol.Biol.Intnl.46. 787-794 (1998)
[Publications] Susumu Tomita: "Interaction of a Neuron-specific Protein Containing PDZ Domains with Alzhemer's Amyloid Precursor Protein" J.Biol.Chem.274. 2243-2254 (1999)
[Publications] Satoshi Watanabe: "Glutamate Induces Cl^- and K^+ Curents in the Olfacotry Interneurons of a Terrestrial Slug" J.Comp.Physiol.A. (in press). (1999)
[Publications] Kanae Ando: "Role of Phsphorylation of Alzheimer's Amyloid Precursor Protein during Neuronal Differentiation" J.Neurosci.(in press). (1999)
[Publications] 桐野豊: "基礎薬学物理化学(改訂版)" 廣川書店(in press), (1999)

1998 Fiscal Year Annual Research Report

モルモット単離小脳を用いたin vitro学習系の開発

Principal Investigator

桐野 豊 東京大学, 大学院・薬学系研究科, 教授 (10012668)

Research Products

[Publications] Ko-ichi Iijima: "Phosphorylated Form of Alzheimer's Amyloid Precursor Protein Exists in Cholinergic Nerve T ermini from Electric Ray Electric Organ" Biochem.J.330. 29-33 (1998)

[Publications] Susumu Tomita: "Cleavage of Alzheimer's Amyloid Precursor Protein by Secretases Occure after O-glycosyla-tion of APP in Protein Secretory Pathway" J.Biol.Chem.273. 6277-6284 (1998)

[Publications] Hirotasu Kojima: "Development of a Fluorescent Indicator for Nitric Oxide Based on the Fluorescein Chromophore" Chem. Pharm. bull.46. 373-375 (1998)

[Publications] Yasushi Satoh: "Lambert-Eaton Syndrome Antibodies Inhibit Acetylcholine Release and P/Q-type Ca^<2+> Channels in Electric Ray Nerve Endings" J.Physiol.508. 427-438 (1998)

[Publications] Tetsuya Kimura: "Behavioral Modulation Indced 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. 4. 365-375 (1998)

[Publications] Tetsuya Kimura: "Optical Recording Analysis of Olfactory Response of the PC Lobe in the Slug Brain" Learning and Memory. 4. 389-400 (1998)

[Publications] Satoshi Watanabe: "Morphological Characterization of the Bursting and Nonbursting Neurons in the Olfactory Center of the Terrestrial Slug Limax marginatus" J.Exp.Biol.201. 925-930 (1998)

[Publications] Yong I.Kim: "An Autoimmune Animal Model of the Lambert-Eaton Syndrom" Ann.N.Y.Acad.Sci. 841. 670-676 (1998)

[Publications] Shigeo Watanabe: "Differential Roles of Two Types of Voltage-gated Ca^<2+> Channels in the Dendrites of Rat Cerebellar Purkinje Neurons" Brain Res.791. 43-55 (1998)

[Publications] Susumu Tomita: "A Basic Amino Acid in the Cytopolasmic Domain of Altzheimer's β-Amyloid Precursor Protein(APP)Is Essential for Cleavage of APP at the α-site" J.Biol.Chem.273. 19304-19310 (1998)

[Publications] Hirotatsu Kojima: "Detection and Imaging of Nitric Oxide with Novel Fluorescent Indicators:Diaminofluoresceins" Anal.Chem.70. 2446-2453 (1998)

[Publications] 桐野 豊: "学習記憶研究の戦略" 放射線医学. 38. 174-177 (1998)

[Publications] Dong-Seok Lee: "Phospholipid Translocation from Outer to Inner Leafled of Synaptic Vescle membranes Isolated from the Electric Organ of Japanese Electric Ray Narke japonica" J.Biochem.124. 798-803 (1998)

[Publications] Tsuyoshi Inoue: "Selective Optical Recording of the Neural Activity in the Olfactory Center of Land Slug Using a Calcium Indicator Dye" Bioimages. 6. 59-67 (1998)

[Publications] Azusa Nakagawa-Inoue: "Odorant-evoked Increase Cytosolic Free Calcium in Cultured Anntenal Neurons of Blowflies" Zool.Sci.15. 661-666 (1998)

[Publications] Yutaka Hirakura: "Membrane Perturbation by the Alzhemer Amyloid Fragment β25-35 Requires Aggregation" Biochem.Mol.Biol.Intnl.46. 787-794 (1998)

[Publications] Susumu Tomita: "Interaction of a Neuron-specific Protein Containing PDZ Domains with Alzhemer's Amyloid Precursor Protein" J.Biol.Chem.274. 2243-2254 (1999)

[Publications] Satoshi Watanabe: "Glutamate Induces Cl^- and K^+ Curents in the Olfacotry Interneurons of a Terrestrial Slug" J.Comp.Physiol.A. (in press). (1999)

[Publications] Kanae Ando: "Role of Phsphorylation of Alzheimer's Amyloid Precursor Protein during Neuronal Differentiation" J.Neurosci.(in press). (1999)

[Publications] 桐野 豊: "基礎薬学 物理化学(改訂版)" 廣川書店(in press), (1999)

桐野豊東京大学, 大学院・薬学系研究科, 教授 (10012668)

[Publications] 桐野豊: "学習記憶研究の戦略" 放射線医学. 38. 174-177 (1998)

[Publications] 桐野豊: "基礎薬学物理化学(改訂版)" 廣川書店(in press), (1999)