2000 Fiscal Year Annual Research Report

新規アセチルコリン合成酵素の脳ニューロン局在と投射様式の解析

Research Project

Project/Area Number	12480225
Research Institution	Shiga University of Medical Science
Principal Investigator	木村宏滋賀医科大学, 分子神経科学研究センター, 教授 (40079736)
Co-Investigator(Kenkyū-buntansha)	相見良成滋賀医科大学, 分子神経科学研究センター, 助手 (20231756) 安原治滋賀医科大学, 分子神経科学研究センター, 助教授 (80239772) 遠山育夫滋賀医科大学, 分子神経科学研究センター, 教授 (20207533)
Keywords	コリンアセチル基転移酵素 / アセチルコリン / コリン神経 / 隆起乳頭体核 / 免疫組織化学 / スプライシング / mRNA / 遺伝子発現
Research Abstract	コリンアセチル基転移酵素(アセチルコリンの合成酵素)は、コリン作動性神経の最も信頼できるマーカーと考えられてきたが、そのアイソザイムが存在するという報告はなかった。本研究者らは最近、コリンアセチル基転移酵素のスプライス・バリアント(pChATと命名)が存在することを発見した。このpChATは基質結合ドメインおよび酵素活性ドメインの両者を備えており、既知の酵素と同様にアセチルコリン産生能をもつアイソザイムである。したがって、pChATはコリン作動性神経の新しいマーカーであると云える。ラットの全身臓器を免疫組織化学法を用いて調査したところ、pChATは主として末梢コリン神経系に分布することを見出し、報告してきた。その後さらに詳細にラット脳を調べたところ、視床下部の隆起乳頭体(大細胞性)核を構成する少なくとも半数のニューロンにpChAT免疫陽性反応が局在することに気付いた。本研究では「隆起乳頭体核-大脳皮質路」におけるコリン作動性神経の関与について検索し12年度には次の結果を得た。 (1)遺伝子解析脳においてRT-PCR法を実施し、そのPCR産物について遺伝子配列を決定たところ、翼口蓋神経節で同定したものと同一配列のmRNAに加え、約100 bpほど長いスプライシング・バリアントの存在を見出した。現在、この配列により翻訳される蛋白を培養細胞に発現させ、その精製を行っているところである。 (2)組織化学的解析翼口蓋神経節で同定したpChATに対する抗体を用いて、ラット隆起乳頭体核に焦点をあて、軸索投射追跡法とを組合せ検索したところ、大脳皮質へ広範な投射を行うことが検証できた。さらに、「隆起乳頭体核-大脳皮質路」に存在するとされる神経伝達関連物質について免疫二重染色法で調べたところ、ヒスタミン合成酵素およびアデノシンデアミネースと、種々の組合せで共存することが明らかとなった。

Research Products
(18 results)

All Other

All Publications (18 results)

[Publications] Suzuki M: "C-type natriuretic peptide-like immunoreactivity in the rat inner ear."Hear.Res.. 139. 51-58 (2000)
[Publications] Park M: "Ontogeny of (D-Ala^2)-deltorphin I-like immunoreactive neurones in foetal rat brain."J.Chem.Neuroanat.. 18. 11-22 (2000)
[Publications] Kitamura Y: "Interleukin-4-inhibited mRNA expression in mixed rat glial and in isolated microglial cultures."J.Neuroimmunology. 106. 95-104 (2000)
[Publications] Nakajima K: "Immunohistochemical demonstration of choline acetyltransferase of a peripheral type (pChAT) in the enteric nervous system of rats."J.Chem.Neuroanat.. 18. 31-40 (2000)
[Publications] Tooyama I: "A protein encoded by an alternative splice variant of choline acetyltransferase mRNA is localized preferentially in peripheral nerve cells and fibers."J.Chem.Neuroanat.. 17. 217-226 (2000)
[Publications] Nakagawa E: "Enhancement of progenitor cell division in the dentate gyrus triggered by initial limbic seizures in rat models of epilepsy."Epilepsia. 41. 10-18 (2000)
[Publications] Kimura H: "Peripheral type choline acetyltransferase : molecular neuroanatomy."Ital.J.Anat.Embryol.. 104. S1 (1999)
[Publications] Nakanishi Y: "Immunohistochemical localization of choline acetyltransferase of a peripheral type in the rat larynx."J.Chem.Neuroanat.. 17. 21-32 (1999)
[Publications] Matsuoka Y: "Kainic acid-induced inducible cyclooxygenase and c-Jun phosphorylation in the rat hippocampal formation."Brain Research. 836. 213-217 (1999)
[Publications] Kitano H: "Regulation of inner ear fluid in the rat by vasopressin."NeuroReport. 10. 1205-1207 (1999)
[Publications] Matsukawa N: "Increased expression of hippocampal cholinergic neurostimulating peptide-related compounds and their messenger mas in the hippocampus of aged senescence-accelerated mice."Neuroscience. 88. 79-92 (1999)
[Publications] Sasaki K: "Effect of acidic fibroblast growth factor fragment analog on learning and memory, and medial septum cholinergic neurons in senescence accelerated mice."Neuroscience. 92. 1287-1294 (1999)
[Publications] Tooyama I: "Microglial reaction and neuronal death in the hippocampus of rat models of epilepsy."Neuropathol.. 19. 203-208 (1999)
[Publications] Yuasa T: "Overproduction of gamma interferon in B/Jas inbred Rabbits with herpes simplex virus encephalitis."Microbiol.Immunol.. 43. 365-371 (1999)
[Publications] Matsuoka Y: "Interferon-γ plus lipopolysaccharide induction of delayed neuronal apoptosis in rat hippocampus."Neurochem.Int.. 34. 91-99 (1999)
[Publications] Kitano H: "Expression of inducible nitric oxide synthase in human thyroid papillary carcinoma."Thyroid. 9. 113-117 (1999)
[Publications] Aisaka A: "Two modes of corticospinal reinnervation occur close to spinal targets following unilateral lesion of the motor cortex in neonatal hamsters."Neuroscience. 90. 53-67 (1999)
[Publications] Kitamura Y: "Hydrogen peroxide-induced apoptosis mediated by p53 protein in glial cells."Glia. 25. 154-164 (1999)

2000 Fiscal Year Annual Research Report

新規アセチルコリン合成酵素の脳ニューロン局在と投射様式の解析

Principal Investigator

木村 宏 滋賀医科大学, 分子神経科学研究センター, 教授 (40079736)

Research Products

[Publications] Suzuki M: "C-type natriuretic peptide-like immunoreactivity in the rat inner ear."Hear.Res.. 139. 51-58 (2000)

[Publications] Park M: "Ontogeny of (D-Ala^2)-deltorphin I-like immunoreactive neurones in foetal rat brain."J.Chem.Neuroanat.. 18. 11-22 (2000)

[Publications] Kitamura Y: "Interleukin-4-inhibited mRNA expression in mixed rat glial and in isolated microglial cultures."J.Neuroimmunology. 106. 95-104 (2000)

[Publications] Nakajima K: "Immunohistochemical demonstration of choline acetyltransferase of a peripheral type (pChAT) in the enteric nervous system of rats."J.Chem.Neuroanat.. 18. 31-40 (2000)

[Publications] Tooyama I: "A protein encoded by an alternative splice variant of choline acetyltransferase mRNA is localized preferentially in peripheral nerve cells and fibers."J.Chem.Neuroanat.. 17. 217-226 (2000)

[Publications] Nakagawa E: "Enhancement of progenitor cell division in the dentate gyrus triggered by initial limbic seizures in rat models of epilepsy."Epilepsia. 41. 10-18 (2000)

[Publications] Kimura H: "Peripheral type choline acetyltransferase : molecular neuroanatomy."Ital.J.Anat.Embryol.. 104. S1 (1999)

[Publications] Nakanishi Y: "Immunohistochemical localization of choline acetyltransferase of a peripheral type in the rat larynx."J.Chem.Neuroanat.. 17. 21-32 (1999)

[Publications] Matsuoka Y: "Kainic acid-induced inducible cyclooxygenase and c-Jun phosphorylation in the rat hippocampal formation."Brain Research. 836. 213-217 (1999)

[Publications] Kitano H: "Regulation of inner ear fluid in the rat by vasopressin."NeuroReport. 10. 1205-1207 (1999)

[Publications] Matsukawa N: "Increased expression of hippocampal cholinergic neurostimulating peptide-related compounds and their messenger mas in the hippocampus of aged senescence-accelerated mice."Neuroscience. 88. 79-92 (1999)

[Publications] Sasaki K: "Effect of acidic fibroblast growth factor fragment analog on learning and memory, and medial septum cholinergic neurons in senescence accelerated mice."Neuroscience. 92. 1287-1294 (1999)

[Publications] Tooyama I: "Microglial reaction and neuronal death in the hippocampus of rat models of epilepsy."Neuropathol.. 19. 203-208 (1999)

[Publications] Yuasa T: "Overproduction of gamma interferon in B/Jas inbred Rabbits with herpes simplex virus encephalitis."Microbiol.Immunol.. 43. 365-371 (1999)

[Publications] Matsuoka Y: "Interferon-γ plus lipopolysaccharide induction of delayed neuronal apoptosis in rat hippocampus."Neurochem.Int.. 34. 91-99 (1999)

[Publications] Kitano H: "Expression of inducible nitric oxide synthase in human thyroid papillary carcinoma."Thyroid. 9. 113-117 (1999)

[Publications] Aisaka A: "Two modes of corticospinal reinnervation occur close to spinal targets following unilateral lesion of the motor cortex in neonatal hamsters."Neuroscience. 90. 53-67 (1999)

[Publications] Kitamura Y: "Hydrogen peroxide-induced apoptosis mediated by p53 protein in glial cells."Glia. 25. 154-164 (1999)

木村宏滋賀医科大学, 分子神経科学研究センター, 教授 (40079736)