{"created":"2023-07-27T06:58:44.170946+00:00","id":57713,"links":{},"metadata":{"_buckets":{"deposit":"33688ac7-7f01-4886-b45f-ffa78a3f4700"},"_deposit":{"created_by":18,"id":"57713","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"57713"},"status":"published"},"_oai":{"id":"oai:kanazawa-u.repo.nii.ac.jp:00057713","sets":["2812:2813:2833"]},"author_link":["20432","100929"],"item_9_biblio_info_8":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"2002-03-25","bibliographicIssueDateType":"Issued"},"bibliographicPageStart":"2p.","bibliographicVolumeNumber":"1999 – 2000","bibliographic_titles":[{"bibliographic_title":"平成12(2000)年度 科学研究費補助金 基盤研究(C) 研究成果報告書概要"},{"bibliographic_title":"2000 Fiscal Year Final Research Summary","bibliographic_titleLang":"en"}]}]},"item_9_creator_33":{"attribute_name":"著者別表示","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{}],"nameIdentifiers":[{},{},{}]}]},"item_9_description_21":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"1.抗ヒト・シナプス小胞アセチルコリントランスポーター(VAChT)抗体の作成\n組み換え発現ベクターを用いて、ヒトVAChTのカルボキシル基末端の細胞質ドメイン(58個のアミノ酸からなる)を大腸菌に発現させた。これをカラムや電気泳動法を用いて精製し、大腸菌培養液8Lから最終的に約2.9mgのVAChT蛋白を得た。これをラットに免疫し、血清を精製してヒトVAChT抗体(パラフィン包埋標本に対しても免疫組織化学が可能)を作成した。\n2.免疫組織化学とIn situ hybridization法によるヒト中枢神経系におけるコリンアセチルトランスフェラーゼ(ChAT)とVAChTの発現の多様性の検討\nすでにクローニング済みのヒトChATとヒトVAChTをpBSベクターに組み込み、RNAプローブを作成してヒト剖検脳に対しin situ hybridizationを行った。また作成したヒトChATとヒトVAChTに対する抗体も用い免疫組織化学を行った。VAChTの分布はChATの分布と類似していることがわかったが完全には一致せず、また投射型コリン作動性神経細胞は局所回路型コリン作動性神経細胞に比べ、ChAT、VAChTともに強い発現のあることがわかった。さらに、投射型コリン作動性神経細胞の中でも、特にMeynert基底核の神経細胞は常時ChATとVAChTを合成しており、その機能を維持していることが推測された。すなわち、同じようにみえるコリン作動性神経細胞の中で機能蛋白発現に強弱があることが判明した。\n3.Reverse transcription-polymerase chain reaction(RT-PCR)・サザンブロット法によるChAT mRNAの多様性の検討\nヒト剖検脳凍結標本よりmRNAを抽出し、5'非翻訳領域の異なる3種類のChAT mRNAがどの領域で認められるかRT-PCR・サザンブロット法を用いて検討した。尾状核ではR型のChAT mRNAが2種類存在し、N型やM型は検出されなかった。一方、これまでの我々の研究では、脊髄前角にM型のChAT mRNAが検出されたが、N型やR型は検出されていない。従って、尾状核と脊髄前角とではChAT遺伝子の転写機構が異なることが明らかになった。","subitem_description_type":"Abstract"},{"subitem_description":"Choline acetyltransferase (ChAT), an enzyme for biosynthesis of acetylcholine, and vesicular acetylcholine transporter (VAChT), a molecule for the translocation of cytoplasmic acetylcholine into synaptic vesicles, are known to be good markers for cholinergic neurons. In order to investigate whether cholinergic neurons are diverse with relation to localization and function, immunohistochemical and in situ hybridization studies were performed on the human central nervous by using specific antibodies and RNA probes for human ChAT and VAChT.In this study, an antibody to human recombinant VAChT protein was developed, which was applicable to paraffin-embedded tissue sections. The distributions of ChAT mRNA, ChAT protein, VAChT mRNA and VAChT protein were similar, but not exactly the same in the human central nervous system. In addition, cholinergic projection neurons tended to express these molecules more than cholinergic local circuit neurons, which might be related to the different functional levels between the two types of cholinergic neurons. Southern blot analysis following reverse transcription- polymerase chain reaction revealed that the caudate nucleus contained two R-type mRNAs, but not N-or M-type mRNA.Our previous study demonstrated that in the human spinal cord M-type mRNA is much more abundant than R-type mRNA.These results suggest that transcription mechanisms of ChAT are different in cholinergic neurons between the caudate nucleus and spinal cord.","subitem_description_type":"Abstract"}]},"item_9_description_22":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"研究課題/領域番号:11670171, 研究期間(年度):1999 – 2000","subitem_description_type":"Other"},{"subitem_description":"出典：「ヒト中枢神経系におけるコリン作動性ニューロンの多様性」研究成果報告書　課題番号11670171\n（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） \n（ https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-11670171/ ）を加工して作成","subitem_description_type":"Other"}]},"item_9_description_5":{"attribute_name":"提供者所属","attribute_value_mlt":[{"subitem_description":"金沢大学医学部・附属病院","subitem_description_type":"Other"}]},"item_9_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.24517/00063983","subitem_identifier_reg_type":"JaLC"}]},"item_9_relation_28":{"attribute_name":"関連URI","attribute_value_mlt":[{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/search/?kw=70169316"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/search/?kw=70169316","subitem_relation_type_select":"URI"}},{"subitem_relation_name":[{"subitem_relation_name_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-11670171/"}],"subitem_relation_type_id":{"subitem_relation_type_id_text":"https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-11670171/","subitem_relation_type_select":"URI"}}]},"item_9_version_type_25":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2021-09-10"}],"displaytype":"detail","filename":"ME-PR-ODA-Y-kaken 2002-2p.pdf","filesize":[{"value":"83.1 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"ME-PR-ODA-Y-kaken 2002-2p.pdf","url":"https://kanazawa-u.repo.nii.ac.jp/record/57713/files/ME-PR-ODA-Y-kaken 2002-2p.pdf"},"version_id":"9fb6406b-f902-4d23-92a1-6371f0b7098e"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"research report","resourceuri":"http://purl.org/coar/resource_type/c_18ws"}]},"item_title":"ヒト中枢神経系におけるコリン作動性ニューロンの多様性","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"ヒト中枢神経系におけるコリン作動性ニューロンの多様性"},{"subitem_title":"Diversity of cholinergic neurons in the human central nervous system","subitem_title_language":"en"}]},"item_type_id":"9","owner":"18","path":["2833"],"pubdate":{"attribute_name":"公開日","attribute_value":"2021-09-10"},"publish_date":"2021-09-10","publish_status":"0","recid":"57713","relation_version_is_last":true,"title":["ヒト中枢神経系におけるコリン作動性ニューロンの多様性"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-07-27T14:58:37.983531+00:00"}