{"id":{"repo_id":"ajou","oai_identifier":"oai:repository.ajou.ac.kr:201003/4381"},"canonical_url":"https://search.dev.ndltd.org/etd/ajou/oai:repository.ajou.ac.kr:201003/4381","repository":{"repo_id":"ajou","name":"Ajou University","base_url":"http://repository.ajou.ac.kr/oai/request"},"display":{"title":"Regulation of Weibel-Palade Body Exocytosis by alpha-Synuclein in Endothelial Cells","abstract":"a-Synuclein (a-syn) is a small presynaptic protein implicated in the pathogenesis of Parkinson’s disease. Although gathering evidence has been proposed for its physiological roles, the precise roles of a-synuclein and mechanisms remain incompletely understood. a-Synuclein is not only expressed in neuron, but also in vascular endothelium. Endothelial cells contain intracellular granules called Weibel-Palade bodies (WPBs) that contain a number of chemokines, adhesive molecules and inflammatory cytokines. This study explored whether the exocytosis of WPB is regulated by a-synuclein. Results showed that PMA-, thrombin- or forskolin-induced VWF release or translocation of P-selectin from Human Umbilical Vein Endothelial Cells (HUVECs) were inhibited by overexpression of a- and b-synuclein, but not g-synuclein. The overexpression of three point mutants (A30P, A53T and E46K) found in familial Parkinson’s disease inhibited WPB exocytosis similar to that of wild-type a-synuclein. Results also showed that the negative regulation of WPB exocytosis required the N-terminus or NAC region of a-synuclein, but not C-terminal acidic tail, and that a-synuclein affected WPB exocytosis through interference with RalA activation by enhancing the interaction of RalGDS/b-arrestin complexes. Immuno-EM analysis revealed that a-synuclein was localized close to WPB. These findings imply that a-synuclein plays as a negative regulator in WPB exocytosis in endothelial cells.","abstract_html":"a-Synuclein (a-syn) is a small presynaptic protein implicated in the pathogenesis of Parkinson’s disease. Although gathering evidence has been proposed for its physiological roles, the precise roles of a-synuclein and mechanisms remain incompletely understood. a-Synuclein is not only expressed in neuron, but also in vascular endothelium. Endothelial cells contain intracellular granules called Weibel-Palade bodies (WPBs) that contain a number of chemokines, adhesive molecules and inflammatory cytokines. This study explored whether the exocytosis of WPB is regulated by a-synuclein. Results showed that PMA-, thrombin- or forskolin-induced VWF release or translocation of P-selectin from Human Umbilical Vein Endothelial Cells (HUVECs) were inhibited by overexpression of a- and b-synuclein, but not g-synuclein. The overexpression of three point mutants (A30P, A53T and E46K) found in familial Parkinson’s disease inhibited WPB exocytosis similar to that of wild-type a-synuclein. Results also showed that the negative regulation of WPB exocytosis required the N-terminus or NAC region of a-synuclein, but not C-terminal acidic tail, and that a-synuclein affected WPB exocytosis through interference with RalA activation by enhancing the interaction of RalGDS/b-arrestin complexes. Immuno-EM analysis revealed that a-synuclein was localized close to WPB. These findings imply that a-synuclein plays as a negative regulator in WPB exocytosis in endothelial cells.","abstract_has_math":false,"creators":["김, 광수"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["주, 일로","박, 상면","대학원 의생명과학과","105432"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-11-08T01:18:59Z","date_published":"2011-11-08T01:18:59Z","updated_at":"2026-07-24T00:51:36Z","subjects":["혈관내피세포","Weibel-Palade Body","exocytosis","Endothelial cells","알파시누클레인","alpha-Synuclein"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011658","000000011658"],"render_values":[{"text":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011658","href":"http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011658","code":true},{"text":"000000011658","href":null,"code":true}]}]},"links":{"outbound_url":"http://repository.ajou.ac.kr/handle/201003/4381","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["주, 일로","박, 상면","대학원 의생명과학과","105432","김, 광수"]},{"key":"dc:creator","label":"Author","values":["김, 광수"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-11-08T01:18:59Z","2011"]},{"key":"dc:type","label":"Dc Type","values":["Thesis","Theses"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["혈관내피세포","Weibel-Palade Body","exocytosis","Endothelial cells","알파시누클레인","alpha-Synuclein"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://repository.ajou.ac.kr/handle/201003/4381","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011658","000000011658"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["a-Synuclein (a-syn) is a small presynaptic protein implicated in the pathogenesis of Parkinson’s disease. Although gathering evidence has been proposed for its physiological roles, the precise roles of a-synuclein and mechanisms remain incompletely understood. a-Synuclein is not only expressed in neuron, but also in vascular endothelium. Endothelial cells contain intracellular granules called Weibel-Palade bodies (WPBs) that contain a number of chemokines, adhesive molecules and inflammatory cytokines. This study explored whether the exocytosis of WPB is regulated by a-synuclein. Results showed that PMA-, thrombin- or forskolin-induced VWF release or translocation of P-selectin from Human Umbilical Vein Endothelial Cells (HUVECs) were inhibited by overexpression of a- and b-synuclein, but not g-synuclein. The overexpression of three point mutants (A30P, A53T and E46K) found in familial Parkinson’s disease inhibited WPB exocytosis similar to that of wild-type a-synuclein. Results also showed that the negative regulation of WPB exocytosis required the N-terminus or NAC region of a-synuclein, but not C-terminal acidic tail, and that a-synuclein affected WPB exocytosis through interference with RalA activation by enhancing the interaction of RalGDS/b-arrestin complexes. Immuno-EM analysis revealed that a-synuclein was localized close to WPB. These findings imply that a-synuclein plays as a negative regulator in WPB exocytosis in endothelial cells.","Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 5 A. MATERIALS 5 1. Reagents and Antibodies 5 2. Constructs 5 B. METHODS 6 1. Cell Culture and Transfection 6 2. RT-PCR analysis 6 3. Determination of VWF by ELISA 7 4. Leukocyte Adhesion Assay 8 5. Ral Activation Assay 8 6. Confocal Microscopy 9 7. Immuno-Electron Microscopy 9 8. Western Blot and Immunoprecipitation 10 Ⅲ. RESULTS 12 1. Three different kinds of secretagogues induce vWF secretion in HUVECs in a time dependent manners 12 2. -Syn overexpression inhibits three secretagogues-induced VWF secretion in HUVECs 14 3. WPB exocytosis is inhibited by -syn and -syn, but not by -syn overexpression 19 4. -Syn mutants (A30P, A53T and E46K) inhibit WPB exocytosis similar to that of wild-type -syn 19 5. N-terminal or NAC region of a-syn is necessary for the inhibition of PMA-induced VWF release in HUVECs 23 6. Secretagogues-induced RalA activation is suppressed by -syn overexpression 28 7. The interaction between RalGDS and -arrestin is enhanced by -syn overexpression 31 8. -Syn is localized close to WPB in HUVECs 35 9. Exogenously added recombinant -syn also inhibits VWF release in HUVECs 38 Ⅳ. DISCUSSION 41 Ⅴ. CONCLUSION 47 REFERENCES 48 국문 요약 59","Doctor"]},{"key":"dc:title","label":"Title","values":["Regulation of Weibel-Palade Body Exocytosis by alpha-Synuclein in Endothelial Cells","혈관내피세포에서 알파시누클레인 (alpha-Synuclein)에 의한 Weibel-Palade Body의 exocytosis 조절"]}]}],"canonical_facts":{"dc:contributor":["주, 일로","박, 상면","대학원 의생명과학과","105432","김, 광수"],"dc:creator":["김, 광수"],"dc:date":["2011-11-08T01:18:59Z","2011"],"dc:description":["a-Synuclein (a-syn) is a small presynaptic protein implicated in the pathogenesis of Parkinson’s disease. Although gathering evidence has been proposed for its physiological roles, the precise roles of a-synuclein and mechanisms remain incompletely understood. a-Synuclein is not only expressed in neuron, but also in vascular endothelium. Endothelial cells contain intracellular granules called Weibel-Palade bodies (WPBs) that contain a number of chemokines, adhesive molecules and inflammatory cytokines. This study explored whether the exocytosis of WPB is regulated by a-synuclein. Results showed that PMA-, thrombin- or forskolin-induced VWF release or translocation of P-selectin from Human Umbilical Vein Endothelial Cells (HUVECs) were inhibited by overexpression of a- and b-synuclein, but not g-synuclein. The overexpression of three point mutants (A30P, A53T and E46K) found in familial Parkinson’s disease inhibited WPB exocytosis similar to that of wild-type a-synuclein. Results also showed that the negative regulation of WPB exocytosis required the N-terminus or NAC region of a-synuclein, but not C-terminal acidic tail, and that a-synuclein affected WPB exocytosis through interference with RalA activation by enhancing the interaction of RalGDS/b-arrestin complexes. Immuno-EM analysis revealed that a-synuclein was localized close to WPB. These findings imply that a-synuclein plays as a negative regulator in WPB exocytosis in endothelial cells.","Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 5 A. MATERIALS 5 1. Reagents and Antibodies 5 2. Constructs 5 B. METHODS 6 1. Cell Culture and Transfection 6 2. RT-PCR analysis 6 3. Determination of VWF by ELISA 7 4. Leukocyte Adhesion Assay 8 5. Ral Activation Assay 8 6. Confocal Microscopy 9 7. Immuno-Electron Microscopy 9 8. Western Blot and Immunoprecipitation 10 Ⅲ. RESULTS 12 1. Three different kinds of secretagogues induce vWF secretion in HUVECs in a time dependent manners 12 2. -Syn overexpression inhibits three secretagogues-induced VWF secretion in HUVECs 14 3. WPB exocytosis is inhibited by -syn and -syn, but not by -syn overexpression 19 4. -Syn mutants (A30P, A53T and E46K) inhibit WPB exocytosis similar to that of wild-type -syn 19 5. N-terminal or NAC region of a-syn is necessary for the inhibition of PMA-induced VWF release in HUVECs 23 6. Secretagogues-induced RalA activation is suppressed by -syn overexpression 28 7. The interaction between RalGDS and -arrestin is enhanced by -syn overexpression 31 8. -Syn is localized close to WPB in HUVECs 35 9. Exogenously added recombinant -syn also inhibits VWF release in HUVECs 38 Ⅳ. DISCUSSION 41 Ⅴ. CONCLUSION 47 REFERENCES 48 국문 요약 59","Doctor"],"dc:identifier":["http://repository.ajou.ac.kr/handle/201003/4381","http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011658","000000011658"],"dc:language":["en"],"dc:subject":["혈관내피세포","Weibel-Palade Body","exocytosis","Endothelial cells","알파시누클레인","alpha-Synuclein"],"dc:title":["Regulation of Weibel-Palade Body Exocytosis by alpha-Synuclein in Endothelial Cells","혈관내피세포에서 알파시누클레인 (alpha-Synuclein)에 의한 Weibel-Palade Body의 exocytosis 조절"],"dc:type":["Thesis","Theses"]},"updated_at":"2026-07-24T00:51:36Z"}