1996 Fiscal Year Annual Research Report

アルコール依存症の分子機構:神経系の細胞内シグナル伝達機構へのエタノールの影響

Research Project

Project/Area Number	08044264
Research Institution	Hamamatsu University School of Medicine
Principal Investigator	鈴木修浜松医科大学, 医学部, 教授 (70093044)
Co-Investigator(Kenkyū-buntansha)	GORDON Adrie カリフォルニア大学, 神経学教室, 教授 DIAMOND Ivan カリフォルニア大学, 神経学教室, 教授浅井清文名古屋市立大学, 分子医学研究所, 助手 (70212462) 西川正信浜松医科大学, 医学部, 助手 (60262812) 石井晃浜松医科大学, 医学部, 助手 (30252175) 妹尾洋浜松医科大学, 医学部, 助教授 (50236113)
Keywords	アルコール / 培養神経細胞 / プロモーター / アデノシンレセプター / cAMP / プロテインキナーゼA / プロテインキナーゼC / 細胞内局在
Research Abstract	アルコール依存症の動物モデルでの研究は広く行われているが、今回、我々はモデルとして培養神経細胞を用いた。エタノール(EtOH)は種々のレセプターなどに作用し、複数のシグナル伝達経路に影響を及ぼすが、培養細胞の場合、各経路への影響や関与する分子の解析が容易となる。またレセプターcDNAを細胞に導入することで、薬物依存の形成に関与していると思われるレセプターと、EtOHとの相互作用の解析も可能となるまず、我々はレセプター発現用のウイルスプロモーターのEtOHによる影響を検討した。3種類のウイルスプロモーターを用い、レセプターのリガンドとの結合やCATアッセイでEtOHの慢性投与の影響を評価した。一時的にCATを発現させた場合、慢性EtOH刺激はどのプロモーターでもCAT活性を上昇させたが、cDNAが安定に発現するクローンでは、慢性EtOH刺激によるプロモーター活性の変化は、クローン間で著しい差があった。このことは、EtOHによるプロモーター活性の変化が、付近のシスエレメントによって影響を受けることを示す。次に、上記の実験で選択されたクローンを用いて、慢性EtOH刺激によるA_1及びA_2レセプターのcAMP産生に対する影響を検討した。その結果、A_1レセプターは、A_2レセプターと違い、慢性EtOH投与による脱感作を受けないことが明かとなった。次いで、EtOHのシグナル伝達系に対する関与をさらに明らかにするために、プロテインキナーゼの細胞内局在の変化を検討した。慢性EtOH投与は、細胞質内に局在するAキナーゼを核内に移動させた。核内に移動したAキナーゼは、CREBなどの種々の転写因子を活性化すると考えられ、このことが、細胞へのEtOHの多彩な影響を説明する現象の一つと考えられる。また、Cキナーゼのあるアイソザイムは、核周囲に存在していたが、EtOH刺激により細胞質へ移動した、他の乱用薬物のレセプターを刺激した時も、Cキナーゼは同じ挙動を示したことから、この現象は、複数の乱用薬物に共通する重要な現象と考えられ、今後Cキナーゼの細胞内局在の変化を中心に研究を進める予定である。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] O.Suzuki.,et al: "Analytical Toxicology" Forensic Science International. 80・1,2. 137-146 (1996)
[Publications] T.Kumazawa.,et al: "Reduction of pyrroloquinoline quinone by dt-diaphorase from clostridium kluyveri" Biochemical Archives. 12. 201-206 (1996)
[Publications] T.Kumazawa.,et al: "Extraction of local anaesthetics from human blood by direct immersion-solid phase micro extraction(SPME)" Chromatographia. 43・1,2. 59-62 (1996)
[Publications] T.Kumazawa.,et al: "Detection of ethanol in human body fluids by headspace solid-phase micro extraction(SPME)/capillary gas chromatography" Chromatographia. 43・7,8. 393-398 (1996)
[Publications] K.MInakata.,et al: "Arch Toxicol" Effect of dietary paraquat on a rat mutant unable to synthesize. 70. 256-258 (1996)
[Publications] H.Seno.,et al: "Detection of some phenothiazines by headspace solid phase microextraction and gas chromatography" Jpn.J.Forensic Toxicol. 14・1. 30-34 (1996)
[Publications] H.Seno.,et al: "Determination of some carbamate pesticides in human body fluids by headspacesolid phase microextraction and gas chromatography" Jpn.J.Forensic Toxicol. 14・3. 199-203 (1996)
[Publications] H.Seno.,et al: "Detection of benzodiazepines in human urine by direct immersion solid phase micro extraction and gas chromatography" Jpn.J.Forensic Toxicol. 15・1. 16-20 (1997)
[Publications] Akira.Ishii.,et al: "Siple and sensitive detection of phencyclidine in body fruids by gas chromatography with surface ionization detection" Int J Legal Med. 108・5. 244-247 (1996)
[Publications] Akira.Ishii.,et al: "Sensitive quantitation of clozapine in body fruids by gas chromatography with surface ionization detection" Jpn.J.Forensic Toxicol. 14・3. 215-220 (1996)
[Publications] Akira.Ishii.,et al: "Simple clean-up methamphetamine and amphetamine in human urine by direct-immersion solid phase micro extraction(DI-SPME)" Jpn.J.Forensic Toxicol. 14・3. 228-232 (1996)
[Publications] Akira.Ishii.,et al: "Simple extraction of phencyclidine from human body fruids by headspace solid-phase microextraction(SPME)" Chromatographia. 43・5,6. 331-334 (1996)
[Publications] Akira.Ishii.,et al: "A simple and sensitive quantitation of N,N-Dimethyltryptamine by gas chromatography with surfase ionization detection" Journal of Analytical Toxicology. 21・1. 36-40 (1997)
[Publications] X.-P.Lee.,et al: "Detection of organophosphate pesticides in human body fruids by headspace solid-phasemicroextraction(SPME) and capillary gas chromatography with nitrogen-phosphorus detection" Chromatographia. 42・3,4. 135-140 (1996)
[Publications] M.Nishikawa.,et al: "The ARS-independent synthesis of DNA complementary to transfer strand in arecipient yeast during transkingdomconjugation between Escherichia coli and Saccharomyces cerevisiae" Nucleic Acids Symposium Series. 35. 273-274 (1996)
[Publications] K.Watanabe.,et al: "Simultaneous determination of cocaethylene and cocaine in blood gas chromatography with surface ionization detection" Chromatographia. 44・1,2. 55-58 (1997)
[Publications] I.Isobe.,et al: "Astrocytic contributions to blood-brain barrier(BBB) formation by endothelial cells:A possible use of aortic endothelial cell for in vitro BBB model" Neurochem Int. 28・5. 523-533 (1996)
[Publications] T.Fuse.,et al: "Heat shock-meditated cell cycle arrest is accompanied by induction of p21 CKI" Biochem. Biophys. Res. Commum. 225・3. 759-763 (1996)
[Publications] K.Matsukawa.,et al: "Tissue distribution of human gliostatin/platelet-derived endothelial cell growth factor(PD-ECGF) and its drug-induced expression" Biochemica et Biophysica Acta. 1371. 71-82 (1996)
[Publications] H.Mizno.,et al: "Glucocorticoid induced the expression of mRNA and the secretion of lipocortin 1 in rat astrocytoma cells" Brain Reserch. 746. 256-268 (1997)
[Publications] Ivan Diamond.,et aal: "Cellular and molecular neuroscience of alcoholism" Physiological Reviews. 77・1. (1997)
[Publications] Douglas P. Dohrman.,et al: "Ethanol causes translocation of cAMP-dependent protein kinase catalytic subunit to nucleus" Cell Biology. 93. 10217-10221 (1996)
[Publications] Imogen R. Coe.,et al: "The role of protein kinase C in cellular tolerance to ethanol" The Journal of Biological Chemistry. 271・46. 29468-29274 (1996)
[Publications] Imogen R. Coe.,et al: "Activation of a cyclic AMP-Dependent protein kinase reverses tolerance of nucleoside transporter to ethanol^1" The Journal of Pharmacology and Experimental Therapeutics. 276・2. 365-369 (1996)

1996 Fiscal Year Annual Research Report

アルコール依存症の分子機構:神経系の細胞内シグナル伝達機構へのエタノールの影響

Principal Investigator

鈴木 修 浜松医科大学, 医学部, 教授 (70093044)

Research Products

[Publications] O.Suzuki.,et al: "Analytical Toxicology" Forensic Science International. 80・1,2. 137-146 (1996)

[Publications] T.Kumazawa.,et al: "Reduction of pyrroloquinoline quinone by dt-diaphorase from clostridium kluyveri" Biochemical Archives. 12. 201-206 (1996)

[Publications] T.Kumazawa.,et al: "Extraction of local anaesthetics from human blood by direct immersion-solid phase micro extraction(SPME)" Chromatographia. 43・1,2. 59-62 (1996)

[Publications] T.Kumazawa.,et al: "Detection of ethanol in human body fluids by headspace solid-phase micro extraction(SPME)/capillary gas chromatography" Chromatographia. 43・7,8. 393-398 (1996)

[Publications] K.MInakata.,et al: "Arch Toxicol" Effect of dietary paraquat on a rat mutant unable to synthesize. 70. 256-258 (1996)

[Publications] H.Seno.,et al: "Detection of some phenothiazines by headspace solid phase microextraction and gas chromatography" Jpn.J.Forensic Toxicol. 14・1. 30-34 (1996)

[Publications] H.Seno.,et al: "Determination of some carbamate pesticides in human body fluids by headspacesolid phase microextraction and gas chromatography" Jpn.J.Forensic Toxicol. 14・3. 199-203 (1996)

[Publications] H.Seno.,et al: "Detection of benzodiazepines in human urine by direct immersion solid phase micro extraction and gas chromatography" Jpn.J.Forensic Toxicol. 15・1. 16-20 (1997)

[Publications] Akira.Ishii.,et al: "Siple and sensitive detection of phencyclidine in body fruids by gas chromatography with surface ionization detection" Int J Legal Med. 108・5. 244-247 (1996)

[Publications] Akira.Ishii.,et al: "Sensitive quantitation of clozapine in body fruids by gas chromatography with surface ionization detection" Jpn.J.Forensic Toxicol. 14・3. 215-220 (1996)

[Publications] Akira.Ishii.,et al: "Simple clean-up methamphetamine and amphetamine in human urine by direct-immersion solid phase micro extraction(DI-SPME)" Jpn.J.Forensic Toxicol. 14・3. 228-232 (1996)

[Publications] Akira.Ishii.,et al: "Simple extraction of phencyclidine from human body fruids by headspace solid-phase microextraction(SPME)" Chromatographia. 43・5,6. 331-334 (1996)

[Publications] Akira.Ishii.,et al: "A simple and sensitive quantitation of N,N-Dimethyltryptamine by gas chromatography with surfase ionization detection" Journal of Analytical Toxicology. 21・1. 36-40 (1997)

[Publications] X.-P.Lee.,et al: "Detection of organophosphate pesticides in human body fruids by headspace solid-phasemicroextraction(SPME) and capillary gas chromatography with nitrogen-phosphorus detection" Chromatographia. 42・3,4. 135-140 (1996)

[Publications] M.Nishikawa.,et al: "The ARS-independent synthesis of DNA complementary to transfer strand in arecipient yeast during transkingdomconjugation between Escherichia coli and Saccharomyces cerevisiae" Nucleic Acids Symposium Series. 35. 273-274 (1996)

[Publications] K.Watanabe.,et al: "Simultaneous determination of cocaethylene and cocaine in blood gas chromatography with surface ionization detection" Chromatographia. 44・1,2. 55-58 (1997)

[Publications] I.Isobe.,et al: "Astrocytic contributions to blood-brain barrier(BBB) formation by endothelial cells:A possible use of aortic endothelial cell for in vitro BBB model" Neurochem Int. 28・5. 523-533 (1996)

[Publications] T.Fuse.,et al: "Heat shock-meditated cell cycle arrest is accompanied by induction of p21 CKI" Biochem. Biophys. Res. Commum. 225・3. 759-763 (1996)

[Publications] K.Matsukawa.,et al: "Tissue distribution of human gliostatin/platelet-derived endothelial cell growth factor(PD-ECGF) and its drug-induced expression" Biochemica et Biophysica Acta. 1371. 71-82 (1996)

[Publications] H.Mizno.,et al: "Glucocorticoid induced the expression of mRNA and the secretion of lipocortin 1 in rat astrocytoma cells" Brain Reserch. 746. 256-268 (1997)

[Publications] Ivan Diamond.,et aal: "Cellular and molecular neuroscience of alcoholism" Physiological Reviews. 77・1. (1997)

[Publications] Douglas P. Dohrman.,et al: "Ethanol causes translocation of cAMP-dependent protein kinase catalytic subunit to nucleus" Cell Biology. 93. 10217-10221 (1996)

[Publications] Imogen R. Coe.,et al: "The role of protein kinase C in cellular tolerance to ethanol" The Journal of Biological Chemistry. 271・46. 29468-29274 (1996)

[Publications] Imogen R. Coe.,et al: "Activation of a cyclic AMP-Dependent protein kinase reverses tolerance of nucleoside transporter to ethanol^1" The Journal of Pharmacology and Experimental Therapeutics. 276・2. 365-369 (1996)

鈴木修浜松医科大学, 医学部, 教授 (70093044)