{"created":"2023-06-19T10:26:32.257839+00:00","id":5125,"links":{},"metadata":{"_buckets":{"deposit":"64448d56-bd2d-47f6-8b18-49acce9d8043"},"_deposit":{"created_by":18,"id":"5125","owners":[18],"pid":{"revision_id":0,"type":"depid","value":"5125"},"status":"published"},"_oai":{"id":"oai:muroran-it.repo.nii.ac.jp:00005125","sets":["41:227"]},"author_link":["22747"],"item_81_date_granted_17":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"2014-03-24"}]},"item_81_degree_grantor_10":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_language":"ja","subitem_degreegrantor_name":"室蘭工業大学"},{"subitem_degreegrantor_language":"en","subitem_degreegrantor_name":"Muroran Institute of Technology"}],"subitem_degreegrantor_identifier":[{"subitem_degreegrantor_identifier_name":"10103","subitem_degreegrantor_identifier_scheme":"kakenhi"}]}]},"item_81_degree_name_11":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士（工学）","subitem_degreename_language":"ja"}]},"item_81_description_25":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_description":"application/pdf","subitem_description_type":"Other"}]},"item_81_description_7":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"生物は周りの環境の情報を得る手段の一つとして嗅覚を備えている。嗅覚では匂い分子を嗅覚受容体が受容して，その情報は嗅球へと伝達され，匂いとして認識される。我々はアカハライモリ （Cynops pyrrhogaster）の嗅組織 cDNA ライブラリーから 2種のタンパク質遺伝子 Cp-lip1 と Cp-lip2 を単離した。in situ hybridizationにより両遺伝子の発現箇所を調べた結果，それぞれ分布は異なるがボウマン腺を構成する細胞で発現していた。これらのタンパク質はリポカリンスーパーファミリーに属する。リポカリンタンパク質は多くの場合，疎水性低分子化合物を結合する。嗅組織から単離されるリポカリンタンパク質として Odorant binding protein （OBP） が知られている。Cp-Lip1, 2 タンパク質の結合特性を解析した結果，匂い分子結合能を有することが明らかとなった。また，両タンパク質の間で匂い分子結合特性が異なっていた。嗅神経細胞の応答感度は in vivo において nM から pM のレベルの感受性を示す。しかし，in vitro では μM レベルの感受性を示し，感度が異なることが報告されている。本研究では，Cp-Lip1, 2 タンパク質がこの感度の差異に関係しているのではないかと考え，これらのタンパク質の嗅組織における分布と嗅神経細胞の匂い応答性に与える影響について調べた。本論文の第1章では，嗅覚システムとリポカリンタンパク質についてまとめた。また，本研究の目的を述べた。第2章では，本研究で用いた分子生物学的手法や組織化学的手法について述べた。第3章では，Cp-Lip1 タンパク質と Cp-Lip2 タンパク質に対する抗血清の作製と評価を行った結果を示した。免疫組織化学的染色には目的タンパク質を検出するために抗体が必要となる。精製したCp-Lip1, 2 タンパク質をマウスに免疫し，抗血清を作製した。これらの抗血清の反応性ついて，精製した Cp-Lip1, 2 タンパク質やアカハライモリ嗅組織から抽出したタンパク質を用いたウエスタンブロット法により評価した。その結果，組織から抽出したタンパク質において特異的に Cp-Lip1 タンパク質，またはCp-Lip2 タンパク質を検出可能な抗血清が得られた。第4章では，まずウエスタンブロッティングによりアカハライモリ各組織における Cp-Lip1, 2 タンパク質の分布を調べた。また，免疫組織化学的染色にて嗅組織内でのより詳細な分布を解析した結果を示した。その結果，両タンパク質は嗅組織のみに分布していることを明らかにした。嗅組織では共にボウマン腺と嗅上皮表層に多く見られたが，その分布はそれぞれ異なっていた。Cp-Lip2 タンパク質は嗅組織全体に幅広く分布していた。一方，Cp-Lip1 タンパク質は Cp-Lip2 タンパク質と比較して狭い範囲に局在していることが明らかとなった。また，嗅上皮表層では両者共に嗅繊毛部分に見られた。第5章では，Cp-Lip1 タンパク質存在下と非存在下における嗅神経細胞の匂い応答性を解析した結果を示した。Cp-Lip1 タンパク質と結合親和性の高い4種の匂い分子を混合した溶液で嗅神経細胞を刺激した。10 mM 匂い分子混合溶液では，Cp-Lip1 タンパク質非存在下において嗅神経細胞の興奮が観察されたが，1 mM 匂い分子混合溶液では興奮が観察されなかった。しかし，Cp-Lip1 タンパク質存在下においては， 1 mM 匂い分子混合溶液で刺激した際にも嗅神経細胞の興奮が観察された。このことから Cp-Lip1 が嗅神経細胞の匂い応答性を変化させることが示された。第6章では，本論文の内容を総括した。","subitem_description_language":"ja","subitem_description_type":"Abstract"},{"subitem_description":"Olfactory system is one of the system which of perceive environmental information. Odorants is received by olfactory receptor. And then, the information of the odorants is transmitted to olfactory bulb. We constructed acDNA library from olfactory epithelium of the Japanese common newt, Cynops pyrrhogaster. Two types of lipocalin genes (Cp-lip1 and Cp-lip2) were isolated from the cDNA library. The distribution of cells that express Cp-lip1 and Cp-lip2 genes was elucidated by in situ hybridization. Two genes were expressed in Bowman’s gland. Cp-Lip1 and Cp-Lip2 are a member of a lipocalin super family. Generally, lipocalin binds hydrophobic compounds. Odorant binding protein (OBP) is known as an olfactory specific lipocalin. Cp-Lip1 and Cp-Lip2 had odorant binding capacity. Odorant binding properties were different between two proteins. Olfactory receptor neurons show sensibility to odorants in range of μM in vitro. In contrast, glomeruli can response to lower concentration odorants (range of pM to nM) in vivo. In this study, I investigated distribution of Cp-Lip1 and Cp-Lip2 in olfactory epithelium of the common newt. Moreover, I found difference of olfactory response of olfactory receptor neurons with or without Cp-Lip1.The contents of this thesis were shown below.Chapter1: Introduction and purpose of this study. This chapter described the olfactory system and lipocalin.Chapter2: Materials and methods.Chapter3: Production and assessment of anti-Cp-Lip1 antiserum and anti-Cp-Lip1 antiserum. These antiserums were produced by immunizing mice. These antiserums were assessed by western blotting with recombinant Cp-Lip1, Cp-Lip2 and proteins included in olfactory epithelium of common newt. As a result, I obtained antiserums that could specifically detect Cp-Lip1 or Cp-Lip2.Chapter4: The distribution of Cp-Lip1 and Cp-Lip2 was investigated in the common newt by western blotting and by immunohistochemistry. These proteins were expressed in Bowman’s glands and located in the surface of olfactory epithelium. However, the distribution was different between Cp-Lip1 and Cp-Lip2. The distribution of Cp-Lip1 was localized. In contrast, Cp-Lip2 distributed widely within olfactory epithelium.Chapter5: I observed difference of olfactory response of olfactory receptorneurons with or without Cp-Lip1. Odorant mixture utilized in this study was a mixture of 4 odorants with a high affinity to Cp-Lip1. Olfactory receptor neurons showed olfactory response at 10 mM odorant mixture. However, it didn’t show olfactory response to 1 mM odorant mixture. When odorant mixture was pre-incubated with Cp-Lip1, olfactory receptor neurons showed olfactory response. In this study, I found that the sensitivity of olfactory response was increased by Cp-Lip1.Chapter6: Summary.","subitem_description_language":"en","subitem_description_type":"Abstract"}]},"item_81_dissertation_number_13":{"attribute_name":"学位授与番号","attribute_value_mlt":[{"subitem_dissertationnumber":"甲第361号"}]},"item_81_identifier_registration":{"attribute_name":"ID登録","attribute_value_mlt":[{"subitem_identifier_reg_text":"10.15118/00005116","subitem_identifier_reg_type":"JaLC"}]},"item_81_subject_9":{"attribute_name":"日本十進分類法","attribute_value_mlt":[{"subitem_subject":"464","subitem_subject_scheme":"NDC"}]},"item_81_text_12":{"attribute_name":"学位の種別","attribute_value_mlt":[{"subitem_text_language":"ja","subitem_text_value":"課程博士"}]},"item_81_text_14":{"attribute_name":"報告番号","attribute_value_mlt":[{"subitem_text_language":"ja","subitem_text_value":"甲第361号"}]},"item_81_text_15":{"attribute_name":"学位記番号","attribute_value_mlt":[{"subitem_text_language":"ja","subitem_text_value":"博甲第361号"}]},"item_81_version_type_24":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_access_right":{"attribute_name":"アクセス権","attribute_value_mlt":[{"subitem_access_right":"open access","subitem_access_right_uri":"http://purl.org/coar/access_right/c_abf2"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorAffiliations":[{"affiliationNameIdentifiers":[],"affiliationNames":[{"affiliationName":""}]}],"creatorNames":[{"creatorName":"杉本, 弘文","creatorNameLang":"ja"},{"creatorName":"SUGIMOTO, Hirofumi","creatorNameLang":"en"},{"creatorName":"スギモト, ヒロフミ","creatorNameLang":"ja-Kana"}],"familyNames":[{},{},{}],"givenNames":[{},{},{}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2016-02-15"}],"displaytype":"detail","filename":"A361.pdf","filesize":[{"value":"3.3 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"A361","objectType":"fulltext","url":"https://muroran-it.repo.nii.ac.jp/record/5125/files/A361.pdf"},"version_id":"e0ab0ea3-302b-4d9f-9b27-e757cc6c5280"},{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2016-02-15"}],"displaytype":"detail","filename":"A361_summary.pdf","filesize":[{"value":"373.9 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"A361_summary","objectType":"abstract","url":"https://muroran-it.repo.nii.ac.jp/record/5125/files/A361_summary.pdf"},"version_id":"40b80ede-faa8-40ff-b34f-4261db227e58"}]},"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":"doctoral thesis","resourceuri":"http://purl.org/coar/resource_type/c_db06"}]},"item_title":"アカハライモリ嗅組織特異的リポカリンタンパク質の組織化学的研究","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"アカハライモリ嗅組織特異的リポカリンタンパク質の組織化学的研究","subitem_title_language":"ja"}]},"item_type_id":"81","owner":"18","path":["227"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2014-06-26"},"publish_date":"2014-06-26","publish_status":"0","recid":"5125","relation_version_is_last":true,"title":["アカハライモリ嗅組織特異的リポカリンタンパク質の組織化学的研究"],"weko_creator_id":"18","weko_shared_id":-1},"updated":"2023-12-25T01:20:59.164799+00:00"}