Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis
The mechanism of glucose-induced biphasic insulin release is unknown. We used total internal reflection fluorescence (TIRF) imaging analysis to reveal the process of first- and second-phase insulin exocytosis in pancreatic β cells. This analysis showed that previously docked insulin granules fused a...
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| Veröffentlicht in: | The Journal of cell biology 2007-05, Vol.177 (4), p.695-705 |
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| creator | Ohara-Imaizumi, Mica Fujiwara, Tomonori Nakamichi, Yoko Okamura, Tadashi Akimoto, Yoshihiro Kawai, Junko Matsushima, Satsuki Kawakami, Hayato Watanabe, Takashi Akagawa, Kimio Nagamatsu, Shinya |
| description | The mechanism of glucose-induced biphasic insulin release is unknown. We used total internal reflection fluorescence (TIRF) imaging analysis to reveal the process of first- and second-phase insulin exocytosis in pancreatic β cells. This analysis showed that previously docked insulin granules fused at the site of syntaxin (Synt)1A clusters during the first phase; however, the newcomers fused during the second phase external to the Synt1A clusters. To reveal the function of Synt1A in phasic insulin exocytosis, we generated Synt1A-knockout (Synt1A⁻/⁻) mice. Synt1A⁻/⁻ β cells showed fewer previously docked granules with no fusion during the first phase; second-phase fusion from newcomers was preserved. Rescue experiments restoring Synt1A expression demonstrated restoration of granule docking status and fusion events. Inhibition of other syntaxins, Synt3 and Synt4, did not affect second-phase insulin exocytosis. We conclude that the first phase is Synt1A dependent but the second phase is not. This indicates that the two phases of insulin exocytosis differ spatially and mechanistically. |
| doi_str_mv | 10.1083/jcb.200608132 |
| format | Article |
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We used total internal reflection fluorescence (TIRF) imaging analysis to reveal the process of first- and second-phase insulin exocytosis in pancreatic β cells. This analysis showed that previously docked insulin granules fused at the site of syntaxin (Synt)1A clusters during the first phase; however, the newcomers fused during the second phase external to the Synt1A clusters. To reveal the function of Synt1A in phasic insulin exocytosis, we generated Synt1A-knockout (Synt1A⁻/⁻) mice. Synt1A⁻/⁻ β cells showed fewer previously docked granules with no fusion during the first phase; second-phase fusion from newcomers was preserved. Rescue experiments restoring Synt1A expression demonstrated restoration of granule docking status and fusion events. Inhibition of other syntaxins, Synt3 and Synt4, did not affect second-phase insulin exocytosis. We conclude that the first phase is Synt1A dependent but the second phase is not. This indicates that the two phases of insulin exocytosis differ spatially and mechanistically.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.200608132</identifier><identifier>PMID: 17502420</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>United States: The Rockefeller University Press</publisher><subject>Animals ; Antibodies ; Beta cells ; Biochemistry ; Biphasic insulins ; Cell membranes ; Cells ; Cells, Cultured ; Exocytosis ; Exocytosis - physiology ; Fluorescence ; Imaging ; Insulin ; Insulin - metabolism ; Insulin Secretion ; Islets of Langerhans ; Islets of Langerhans - metabolism ; Male ; Mice ; Mice, Knockout ; Microscopy, Fluorescence ; Neurons ; Scientific imaging ; Type 2 diabetes mellitus</subject><ispartof>The Journal of cell biology, 2007-05, Vol.177 (4), p.695-705</ispartof><rights>Copyright 2007 Rockefeller University Press</rights><rights>Copyright Rockefeller University Press May 21, 2007</rights><rights>Copyright © 2007, The Rockefeller University Press 2007</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c524t-1e0444ff51053762712b6715709ca25795db8a35552d23527a0dff7a4e5ac20e3</citedby><cites>FETCH-LOGICAL-c524t-1e0444ff51053762712b6715709ca25795db8a35552d23527a0dff7a4e5ac20e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,777,781,882,27905,27906</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17502420$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ohara-Imaizumi, Mica</creatorcontrib><creatorcontrib>Fujiwara, Tomonori</creatorcontrib><creatorcontrib>Nakamichi, Yoko</creatorcontrib><creatorcontrib>Okamura, Tadashi</creatorcontrib><creatorcontrib>Akimoto, Yoshihiro</creatorcontrib><creatorcontrib>Kawai, Junko</creatorcontrib><creatorcontrib>Matsushima, Satsuki</creatorcontrib><creatorcontrib>Kawakami, Hayato</creatorcontrib><creatorcontrib>Watanabe, Takashi</creatorcontrib><creatorcontrib>Akagawa, Kimio</creatorcontrib><creatorcontrib>Nagamatsu, Shinya</creatorcontrib><title>Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>The mechanism of glucose-induced biphasic insulin release is unknown. We used total internal reflection fluorescence (TIRF) imaging analysis to reveal the process of first- and second-phase insulin exocytosis in pancreatic β cells. This analysis showed that previously docked insulin granules fused at the site of syntaxin (Synt)1A clusters during the first phase; however, the newcomers fused during the second phase external to the Synt1A clusters. To reveal the function of Synt1A in phasic insulin exocytosis, we generated Synt1A-knockout (Synt1A⁻/⁻) mice. Synt1A⁻/⁻ β cells showed fewer previously docked granules with no fusion during the first phase; second-phase fusion from newcomers was preserved. Rescue experiments restoring Synt1A expression demonstrated restoration of granule docking status and fusion events. Inhibition of other syntaxins, Synt3 and Synt4, did not affect second-phase insulin exocytosis. We conclude that the first phase is Synt1A dependent but the second phase is not. This indicates that the two phases of insulin exocytosis differ spatially and mechanistically.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Beta cells</subject><subject>Biochemistry</subject><subject>Biphasic insulins</subject><subject>Cell membranes</subject><subject>Cells</subject><subject>Cells, Cultured</subject><subject>Exocytosis</subject><subject>Exocytosis - physiology</subject><subject>Fluorescence</subject><subject>Imaging</subject><subject>Insulin</subject><subject>Insulin - metabolism</subject><subject>Insulin Secretion</subject><subject>Islets of Langerhans</subject><subject>Islets of Langerhans - metabolism</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Knockout</subject><subject>Microscopy, Fluorescence</subject><subject>Neurons</subject><subject>Scientific imaging</subject><subject>Type 2 diabetes mellitus</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkUtvEzEURi0EomlhyRKwumA35fo1TjaVUMWjUiUW0LXl8VwnjmbsYM8U8u9xlSg8Vl58x8f3-iPkFYMrBkvxfuu6Kw7QwpIJ_oQsmJLQLJmEp2QBwFmzUlydkfNStgAgtRTPyRnTCrjksCDudrTrENfURjvsSyg04wPaodAR3cbGUKbgaB-8x4zRYaEdTj8RI_Uhl6mp93pa0KXYN7uNLUhDLPMQIsVfye2nVJUvyDNfjfjyeF6Q-08fv998ae6-fr69-XDXOMXl1DAEKaX3ioESuuWa8a7VTGlYOcuVXqm-W1qhlOI9F4prC7332kpU1nFAcUGuD97d3I3YO4xTtoPZ5TDavDfJBvNvEsPGrNOD4dBKzmQVvDsKcvoxY5nMGIrDYbAR01yMBsUYMKjg5X_gNs25_mAxnGkGUBeqUHOAXE6lZPSnSRiYx-5M7c6cuqv8m7_H_0Mfy6rA6wOwLVPKp1w8PgdiVfO3h9zbZOw6h2Luv3FgFdC61SDFb83lqG8</recordid><startdate>20070521</startdate><enddate>20070521</enddate><creator>Ohara-Imaizumi, Mica</creator><creator>Fujiwara, Tomonori</creator><creator>Nakamichi, Yoko</creator><creator>Okamura, Tadashi</creator><creator>Akimoto, Yoshihiro</creator><creator>Kawai, Junko</creator><creator>Matsushima, Satsuki</creator><creator>Kawakami, Hayato</creator><creator>Watanabe, Takashi</creator><creator>Akagawa, Kimio</creator><creator>Nagamatsu, Shinya</creator><general>The Rockefeller University Press</general><general>Rockefeller University Press</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20070521</creationdate><title>Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis</title><author>Ohara-Imaizumi, Mica ; Fujiwara, Tomonori ; Nakamichi, Yoko ; Okamura, Tadashi ; Akimoto, Yoshihiro ; Kawai, Junko ; Matsushima, Satsuki ; Kawakami, Hayato ; Watanabe, Takashi ; Akagawa, Kimio ; Nagamatsu, Shinya</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c524t-1e0444ff51053762712b6715709ca25795db8a35552d23527a0dff7a4e5ac20e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Beta cells</topic><topic>Biochemistry</topic><topic>Biphasic insulins</topic><topic>Cell membranes</topic><topic>Cells</topic><topic>Cells, Cultured</topic><topic>Exocytosis</topic><topic>Exocytosis - physiology</topic><topic>Fluorescence</topic><topic>Imaging</topic><topic>Insulin</topic><topic>Insulin - metabolism</topic><topic>Insulin Secretion</topic><topic>Islets of Langerhans</topic><topic>Islets of Langerhans - metabolism</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Knockout</topic><topic>Microscopy, Fluorescence</topic><topic>Neurons</topic><topic>Scientific imaging</topic><topic>Type 2 diabetes mellitus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ohara-Imaizumi, Mica</creatorcontrib><creatorcontrib>Fujiwara, Tomonori</creatorcontrib><creatorcontrib>Nakamichi, Yoko</creatorcontrib><creatorcontrib>Okamura, Tadashi</creatorcontrib><creatorcontrib>Akimoto, Yoshihiro</creatorcontrib><creatorcontrib>Kawai, Junko</creatorcontrib><creatorcontrib>Matsushima, Satsuki</creatorcontrib><creatorcontrib>Kawakami, Hayato</creatorcontrib><creatorcontrib>Watanabe, Takashi</creatorcontrib><creatorcontrib>Akagawa, Kimio</creatorcontrib><creatorcontrib>Nagamatsu, Shinya</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ohara-Imaizumi, Mica</au><au>Fujiwara, Tomonori</au><au>Nakamichi, Yoko</au><au>Okamura, Tadashi</au><au>Akimoto, Yoshihiro</au><au>Kawai, Junko</au><au>Matsushima, Satsuki</au><au>Kawakami, Hayato</au><au>Watanabe, Takashi</au><au>Akagawa, Kimio</au><au>Nagamatsu, Shinya</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>2007-05-21</date><risdate>2007</risdate><volume>177</volume><issue>4</issue><spage>695</spage><epage>705</epage><pages>695-705</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><abstract>The mechanism of glucose-induced biphasic insulin release is unknown. We used total internal reflection fluorescence (TIRF) imaging analysis to reveal the process of first- and second-phase insulin exocytosis in pancreatic β cells. This analysis showed that previously docked insulin granules fused at the site of syntaxin (Synt)1A clusters during the first phase; however, the newcomers fused during the second phase external to the Synt1A clusters. To reveal the function of Synt1A in phasic insulin exocytosis, we generated Synt1A-knockout (Synt1A⁻/⁻) mice. Synt1A⁻/⁻ β cells showed fewer previously docked granules with no fusion during the first phase; second-phase fusion from newcomers was preserved. Rescue experiments restoring Synt1A expression demonstrated restoration of granule docking status and fusion events. Inhibition of other syntaxins, Synt3 and Synt4, did not affect second-phase insulin exocytosis. We conclude that the first phase is Synt1A dependent but the second phase is not. This indicates that the two phases of insulin exocytosis differ spatially and mechanistically.</abstract><cop>United States</cop><pub>The Rockefeller University Press</pub><pmid>17502420</pmid><doi>10.1083/jcb.200608132</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Antibodies Beta cells Biochemistry Biphasic insulins Cell membranes Cells Cells, Cultured Exocytosis Exocytosis - physiology Fluorescence Imaging Insulin Insulin - metabolism Insulin Secretion Islets of Langerhans Islets of Langerhans - metabolism Male Mice Mice, Knockout Microscopy, Fluorescence Neurons Scientific imaging Type 2 diabetes mellitus |
| title | Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis |
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