{"created":"2023-06-20T13:20:59.598407+00:00","id":1071,"links":{},"metadata":{"_buckets":{"deposit":"7d7e57fd-258b-4ef1-8f76-acd681abcd22"},"_deposit":{"created_by":1,"id":"1071","owners":[1],"pid":{"revision_id":0,"type":"depid","value":"1071"},"status":"published"},"_oai":{"id":"oai:ir.soken.ac.jp:00001071","sets":["2:430:22"]},"author_link":["9232","9230","9231"],"item_1_creator_2":{"attribute_name":"著者名","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"井上, 明宏"}],"nameIdentifiers":[{}]}]},"item_1_creator_3":{"attribute_name":"フリガナ","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"イノウエ, アキヒロ"}],"nameIdentifiers":[{}]}]},"item_1_date_granted_11":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"1992-03-16"}]},"item_1_degree_grantor_5":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_name":"総合研究大学院大学"}]}]},"item_1_degree_name_6":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士（学術）"}]},"item_1_description_12":{"attribute_name":"要旨","attribute_value_mlt":[{"subitem_description":"　　　Monoclonal antibody(MAb) is useful tool to study the\n molecular basis of development, construction and function of the\n nervous system. MAb technique can find unknown molecules\n which exhibit characteristic spatial or temporal expression patterns,\npossibly playing significant roles on the nervous system.　　A\n protein antigen recognized by a MAb, designated <u>HPC-1</u>,localized\n in the plasma membrane of the amacrine cell somas and the inner\n plexiform layer (IPL) in rat retinae.　　MAb HPC-1 recognized\n several proteins of about 35 kDa in SDS-PAGE.\n　　　In the chapter I of this thesis, an Mab HPC-1-positive cDNA\n clone, HPC-113, was isolated from a rat hippocampus cDNA library\n constucted in a lambda phage vector,λgt11, which expressed\n β-galactosidase/cDNA fusion protein. The rabbit antiserum raised\n against the β-galactosidase/HPC-113 fusion protein showed the\n almost same characterisics both in immunoblotting and\n immunohistochemical studies as those with MAb HPC-1 in the rat\n retina. Thus it was concluded that HPC-113 coded for the antigen\n molecule(s) recognized by MAb HPC-1. HPC-113 had 894-bp\n nucleotide sequence in the same open reading frame of <i>E</i>. <i>coli</i>\n β-galactosidase gene and followed by a 1326-bp possible\n 3’noncoding sequence, and the calculated molecular weight of the\n deduced amino acid (298 residues) was 33989 Da, implying that\n HPC-113 contains almost the full-length coding region of HPC-1\n mRNA. The hybrophathy profile of the deduced amino acid\n  sequence showed the presence of an obvious hydrophobic region at the\n  carboxy-terminal end, suggisting that HPC-1 antigen is an\n  integranted membrane protein.　　These results were comparable to\n  the results of biochemical and immunohistochemical studies all of\n  which indicated that HPC-1 should be tightly associated with\n  plasma membrane.　　Although HPC-1 antigen sequence had no\n  typical amino(N)-ternminal signal peptide sequence which was\n  required for secretory and membrane proteins, it was suggested that\n  the large N-terminal side was in the extracellular domain, since\n  MAb HPC-1, of which the epitope was in the N-terminal side,\n reacted with living cells.　　Thus, HPC-113 might not include the\n  complete full-length of the coding region of HPC-1 antigen\n  molecule. On the other hand, the <i>in vitro</i> tlanscription/translation\n  product from mithionine, the 11th residue of the 298-deduced\n  amino acid sequence,co-migrated with the lowest band of HPC-1\n  antigens detected in the immunoblot analysis.　　The cDNA probe,\n  the insert of λHPC-113, detected single 2.4-kb mRNA in the RNA\n  blot analysis, and the S1 nuclase protection analysis probed with\n  the coding region of cDNA also indicated that there was only single\n kind of mRNA for HPC-1 antigen.　　Therefore, it might be possible\n that HPC-1 antigen was translated from Met<sup>11</sup> and its heterogeneity\n  was generated by any posttranslational modifications. 　　However,\n sufficient understanding about the HPC-1 antigen mRNA,its\n  primary structure and heterogeneity are not attained at present.\n  Secondary structure prediction analysis revealed that a part of\n  HPC-1 antigen protein formed α-herical strcture with the\n  periodical heptad repeats by hydrophobic amino acids, which was\n  usually seen in fibrous proteins with dimer or trimer coiled-coil\n structures.　　These results implied that HPC-1 molecule might bind\n to other proteins by its intra- or inter-polypeptide chain association\n capacity.　　Although the whole amino acid sequence did not show\n  significant homology to any known proteins so far, a few local\n sequences in the N-terminal side had notable homologies with some\n partial sequences in mouse laminin B1 chain, a subunit of laminin.\n Its well known that Iaminin, an extracellular matrix protein showed\n various biological function in the nervous system.　　Interestingly\n these homologous sequences in laminin B1 were involved in the\n fragments which revealed neurite-outgrowth and/or survival\n promoting activity. \n　　　cDNA clones for bovine HPC-1 antigen were also isolated.\n The longest cDNA colne, BHPC-109, revealed high nucleotide\n sequence homology in the coding region(91.3%), whereas,\n homology of 3’noncoding regions was lower than that of coding\n region. The 5’end was similar to that of HPC-113, indicating that\n reverse transcriptions were stopped at this 5’ portion of both of rat\n and bovine mRNAs of which sequences possessed quite high G-C\n contents.　　The comparison between the deduced amino acid\n sequences of rat and bovine represented remarkable conservation\n(98%), suggesting a physiogical significance of HPC-1 antigen\n through the mammalian evolution.\n　　In the chapter II, tissue distribution of HPC-1 antigenecity and\n its mRNA was studied by biochemical and histochemical methods.\n Immunoblot analysis showed that the antiserum against the fusion\n protein described above detected several proteins is about 35 kDa in\n the neuronal tissues (retina, cerebral contrex, hippocampus,\n cerebellum and spianl cord), but no proteins is detected in the non-\nneuronal tissues (liver, kidney, heart, muscle and adrenal). 　　On\n the immuohistochemistry of rat nervous system, HPC-1 antigen was\n also observed specifically in the nervous system: the matrices of\n cerebral cortex and hippocampus (Particulaly in stratum radiatum);\n molecular layer, membrane of granular cell soma and gromeruli in\n cerebellum; gray matter of spinal cord. However, little staining\n was detetected in white matters of the central nervous tissues.　　The\n RNA blot analysis also indicated nervous system-specific\n expression of HPC-1 mRNA. In the nonneuronal tissue, however,\n the high sensitive RNA polymerase chain reaction assay revealed\n presence small amount of a HPC-1 gene transcript which appeared\n to be closely related to but distinguishable from the neuronal HPC-1\n gene transcript. <i>In stiu</i> hyblidization was performed by the\n nonradioactive ditiction method to identify cellular localization.\n HPC-1 mRNA was present in most of neurons in the central and\n  peripheral nervous systems except for retina. In the retina, signals\n were detected in amacrine cells, and also in ganglion cells which\n HPC-1 immunoreactivity was not present in the soma, suggesting\n selective localization of HPC-1 mRNA in the IPL of the axon\n terminal. Amount of HPC-1 antigen(s) gradually increased in\n accordance with development or the IPL formation in chick retina.\n Considering from accumulation of HPC-1 antigenecity in the\n hippocampal stratum radiatum, cerebellar gromeruli,and retinal\n IPL, HPC-1 antigen may associate to synaptic formation and/or\n maintenance of neurons. \n　　　In conclusion, it was proved that HPC-1 antigen(s) was a\n novel class of membrane protein(s) of 35 kDa with α-helical\n structure containing typical heptad repeats which related to\n association between other polypeptide. A few local sequences had\n notable homology to some partial laminin sequences that were\n included in the fragments baring neurite-outgrowth and/or survival\n promoting activity. HPC-1 antigen(s) was expressed predominantly\n in the neuronal tissues with characteristic localization, such as\n accumulation into synapse-rich regions, but discrepancy between\n immunoreactivity and presence of\n mRNA in subpopulations of neurons should be solved in future.","subitem_description_type":"Other"}]},"item_1_description_7":{"attribute_name":"学位記番号","attribute_value_mlt":[{"subitem_description":"総研大甲第27号","subitem_description_type":"Other"}]},"item_1_select_14":{"attribute_name":"所蔵","attribute_value_mlt":[{"subitem_select_item":"有"}]},"item_1_select_8":{"attribute_name":"研究科","attribute_value_mlt":[{"subitem_select_item":"生命科学研究科"}]},"item_1_select_9":{"attribute_name":"専攻","attribute_value_mlt":[{"subitem_select_item":"20 生理科学専攻"}]},"item_1_text_10":{"attribute_name":"学位授与年度","attribute_value_mlt":[{"subitem_text_value":"1991"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"INOUE, Akihiro","creatorNameLang":"en"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2016-02-17"}],"displaytype":"simple","filename":"甲27_要旨.pdf","filesize":[{"value":"380.3 kB"}],"format":"application/pdf","licensetype":"license_11","mimetype":"application/pdf","url":{"label":"要旨・審査要旨 / Abstract, Screening Result","url":"https://ir.soken.ac.jp/record/1071/files/甲27_要旨.pdf"},"version_id":"332a6b3a-220e-4080-b52c-00c76ea8e60d"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"thesis","resourceuri":"http://purl.org/coar/resource_type/c_46ec"}]},"item_title":"神経細胞膜抗原ＨＰＣ-１のｃＤＮＡクローニング、一次構造解析及び組織発現様式","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"神経細胞膜抗原ＨＰＣ-１のｃＤＮＡクローニング、一次構造解析及び組織発現様式"},{"subitem_title":"cDNA cloning, sequence analysis and tissue distribution of a neuronal cell membrane antigen, HPC-1","subitem_title_language":"en"}]},"item_type_id":"1","owner":"1","path":["22"],"pubdate":{"attribute_name":"公開日","attribute_value":"2010-02-22"},"publish_date":"2010-02-22","publish_status":"0","recid":"1071","relation_version_is_last":true,"title":["神経細胞膜抗原ＨＰＣ-１のｃＤＮＡクローニング、一次構造解析及び組織発現様式"],"weko_creator_id":"1","weko_shared_id":1},"updated":"2023-06-20T14:48:44.387217+00:00"}