Confocal and multiphoton calcium imaging of the enteric nervous system in anesthetized mice
•New transgenic mice co-express G-CaMP6 and mCherry in ENS neurons.•Intestinal movement stabilization by suction enables imaging of ENS activity in living mice.•Spontaneous and evoked activity can be imaged at cellular and subcellular resolutions. Most imaging studies of the enteric nervous system (...
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| Veröffentlicht in: | Neuroscience research 2020-02, Vol.151, p.53-60 |
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| creator | Motegi, Yuki Sato, Masaaki Horiguchi, Kazuhide Ohkura, Masamichi Gengyo-Ando, Keiko Ikegaya, Yuji Fusamae, Yasuyuki Hongo, Yoshie Suzuki, Minoru Ogawa, Koichi Takaki, Miyako Nakai, Junichi |
| description | •New transgenic mice co-express G-CaMP6 and mCherry in ENS neurons.•Intestinal movement stabilization by suction enables imaging of ENS activity in living mice.•Spontaneous and evoked activity can be imaged at cellular and subcellular resolutions.
Most imaging studies of the enteric nervous system (ENS) that regulates the function of the gastrointestinal tract are so far performed using preparations isolated from animals, thus hindering the understanding of the ENS function in vivo. Here we report a method for imaging the ENS cellular network activity in living mice using a new transgenic mouse line that co-expresses G-CaMP6 and mCherry in the ENS combined with the suction-mediated stabilization of intestinal movements. With confocal or two-photon imaging, our method can visualize spontaneous and pharmacologically-evoked ENS network activity in living animals at cellular and subcellular resolutions, demonstrating the potential usefulness for studies of the ENS function in health and disease. |
| doi_str_mv | 10.1016/j.neures.2019.02.004 |
| format | Article |
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Most imaging studies of the enteric nervous system (ENS) that regulates the function of the gastrointestinal tract are so far performed using preparations isolated from animals, thus hindering the understanding of the ENS function in vivo. Here we report a method for imaging the ENS cellular network activity in living mice using a new transgenic mouse line that co-expresses G-CaMP6 and mCherry in the ENS combined with the suction-mediated stabilization of intestinal movements. With confocal or two-photon imaging, our method can visualize spontaneous and pharmacologically-evoked ENS network activity in living animals at cellular and subcellular resolutions, demonstrating the potential usefulness for studies of the ENS function in health and disease.</description><identifier>ISSN: 0168-0102</identifier><identifier>EISSN: 1872-8111</identifier><identifier>DOI: 10.1016/j.neures.2019.02.004</identifier><identifier>PMID: 30790590</identifier><language>eng</language><publisher>Ireland: Elsevier B.V</publisher><subject>Animals ; Calcium - analysis ; Calcium - metabolism ; Confocal microscopy ; Enteric Nervous System - physiology ; Genetically-encoded calcium indicator ; Gut ; In vivo imaging ; Intestines ; Intravital Microscopy - methods ; Male ; Mice ; Mice, Transgenic ; Microscopy, Fluorescence - methods ; Molecular Imaging - methods ; Neurons - metabolism ; Neurons - physiology ; Serotonin ; Serotonin - pharmacology ; Transgenic mice ; Two-photon microscopy</subject><ispartof>Neuroscience research, 2020-02, Vol.151, p.53-60</ispartof><rights>2019 Elsevier B.V. and Japan Neuroscience Society</rights><rights>Copyright © 2019 Elsevier B.V. and Japan Neuroscience Society. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c452t-8067b4705ca56147098556fcbe372764285d498b3098ef304bc01ba27f5cc67d3</citedby><cites>FETCH-LOGICAL-c452t-8067b4705ca56147098556fcbe372764285d498b3098ef304bc01ba27f5cc67d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0168010218305236$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,65309</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30790590$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Motegi, Yuki</creatorcontrib><creatorcontrib>Sato, Masaaki</creatorcontrib><creatorcontrib>Horiguchi, Kazuhide</creatorcontrib><creatorcontrib>Ohkura, Masamichi</creatorcontrib><creatorcontrib>Gengyo-Ando, Keiko</creatorcontrib><creatorcontrib>Ikegaya, Yuji</creatorcontrib><creatorcontrib>Fusamae, Yasuyuki</creatorcontrib><creatorcontrib>Hongo, Yoshie</creatorcontrib><creatorcontrib>Suzuki, Minoru</creatorcontrib><creatorcontrib>Ogawa, Koichi</creatorcontrib><creatorcontrib>Takaki, Miyako</creatorcontrib><creatorcontrib>Nakai, Junichi</creatorcontrib><title>Confocal and multiphoton calcium imaging of the enteric nervous system in anesthetized mice</title><title>Neuroscience research</title><addtitle>Neurosci Res</addtitle><description>•New transgenic mice co-express G-CaMP6 and mCherry in ENS neurons.•Intestinal movement stabilization by suction enables imaging of ENS activity in living mice.•Spontaneous and evoked activity can be imaged at cellular and subcellular resolutions.
Most imaging studies of the enteric nervous system (ENS) that regulates the function of the gastrointestinal tract are so far performed using preparations isolated from animals, thus hindering the understanding of the ENS function in vivo. Here we report a method for imaging the ENS cellular network activity in living mice using a new transgenic mouse line that co-expresses G-CaMP6 and mCherry in the ENS combined with the suction-mediated stabilization of intestinal movements. With confocal or two-photon imaging, our method can visualize spontaneous and pharmacologically-evoked ENS network activity in living animals at cellular and subcellular resolutions, demonstrating the potential usefulness for studies of the ENS function in health and disease.</description><subject>Animals</subject><subject>Calcium - analysis</subject><subject>Calcium - metabolism</subject><subject>Confocal microscopy</subject><subject>Enteric Nervous System - physiology</subject><subject>Genetically-encoded calcium indicator</subject><subject>Gut</subject><subject>In vivo imaging</subject><subject>Intestines</subject><subject>Intravital Microscopy - methods</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Transgenic</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Molecular Imaging - methods</subject><subject>Neurons - metabolism</subject><subject>Neurons - physiology</subject><subject>Serotonin</subject><subject>Serotonin - pharmacology</subject><subject>Transgenic mice</subject><subject>Two-photon microscopy</subject><issn>0168-0102</issn><issn>1872-8111</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMtOwzAQRS0EouXxBwh5ySZh7Lw3SKjiJVViAysWVuJMqKvELnaCVL6eqQosWY01PnPnzmXsQkAsQOTX69ji5DHEEkQVg4wB0gM2F2Uho1IIccjmhJURCJAzdhLCGgCSKk2O2SyBooKsgjl7WzjbOV33vLYtH6Z-NJuVG53l1NNmGrgZ6ndj37nr-LhCjnZEbzS36D_dFHjYhhGJsiSAgYjRfCEpGY1n7Kir-4DnP_WUvd7fvSweo-Xzw9PidhnpNJNjVEJeNGkBma6zXNCjKrMs73SDSSGLPJVl1qZV2ST0gV0CaaNBNLUsukzrvGiTU3a119149zGRCTWYoLHvyRFZVFLsBHMJFaHpHtXeheCxUxtPB_qtEqB2saq12seqdrEqkIpipbHLnw1TM2D7N_SbIwE3ewDpzk-DXgVt0GpsjUc9qtaZ_zd8A-oeiw8</recordid><startdate>202002</startdate><enddate>202002</enddate><creator>Motegi, Yuki</creator><creator>Sato, Masaaki</creator><creator>Horiguchi, Kazuhide</creator><creator>Ohkura, Masamichi</creator><creator>Gengyo-Ando, Keiko</creator><creator>Ikegaya, Yuji</creator><creator>Fusamae, Yasuyuki</creator><creator>Hongo, Yoshie</creator><creator>Suzuki, Minoru</creator><creator>Ogawa, Koichi</creator><creator>Takaki, Miyako</creator><creator>Nakai, Junichi</creator><general>Elsevier B.V</general><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>7X8</scope></search><sort><creationdate>202002</creationdate><title>Confocal and multiphoton calcium imaging of the enteric nervous system in anesthetized mice</title><author>Motegi, Yuki ; Sato, Masaaki ; Horiguchi, Kazuhide ; Ohkura, Masamichi ; Gengyo-Ando, Keiko ; Ikegaya, Yuji ; Fusamae, Yasuyuki ; Hongo, Yoshie ; Suzuki, Minoru ; Ogawa, Koichi ; Takaki, Miyako ; Nakai, Junichi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c452t-8067b4705ca56147098556fcbe372764285d498b3098ef304bc01ba27f5cc67d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Animals</topic><topic>Calcium - analysis</topic><topic>Calcium - metabolism</topic><topic>Confocal microscopy</topic><topic>Enteric Nervous System - physiology</topic><topic>Genetically-encoded calcium indicator</topic><topic>Gut</topic><topic>In vivo imaging</topic><topic>Intestines</topic><topic>Intravital Microscopy - methods</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Transgenic</topic><topic>Microscopy, Fluorescence - methods</topic><topic>Molecular Imaging - methods</topic><topic>Neurons - metabolism</topic><topic>Neurons - physiology</topic><topic>Serotonin</topic><topic>Serotonin - pharmacology</topic><topic>Transgenic mice</topic><topic>Two-photon microscopy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Motegi, Yuki</creatorcontrib><creatorcontrib>Sato, Masaaki</creatorcontrib><creatorcontrib>Horiguchi, Kazuhide</creatorcontrib><creatorcontrib>Ohkura, Masamichi</creatorcontrib><creatorcontrib>Gengyo-Ando, Keiko</creatorcontrib><creatorcontrib>Ikegaya, Yuji</creatorcontrib><creatorcontrib>Fusamae, Yasuyuki</creatorcontrib><creatorcontrib>Hongo, Yoshie</creatorcontrib><creatorcontrib>Suzuki, Minoru</creatorcontrib><creatorcontrib>Ogawa, Koichi</creatorcontrib><creatorcontrib>Takaki, Miyako</creatorcontrib><creatorcontrib>Nakai, Junichi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Neuroscience research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Motegi, Yuki</au><au>Sato, Masaaki</au><au>Horiguchi, Kazuhide</au><au>Ohkura, Masamichi</au><au>Gengyo-Ando, Keiko</au><au>Ikegaya, Yuji</au><au>Fusamae, Yasuyuki</au><au>Hongo, Yoshie</au><au>Suzuki, Minoru</au><au>Ogawa, Koichi</au><au>Takaki, Miyako</au><au>Nakai, Junichi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Confocal and multiphoton calcium imaging of the enteric nervous system in anesthetized mice</atitle><jtitle>Neuroscience research</jtitle><addtitle>Neurosci Res</addtitle><date>2020-02</date><risdate>2020</risdate><volume>151</volume><spage>53</spage><epage>60</epage><pages>53-60</pages><issn>0168-0102</issn><eissn>1872-8111</eissn><abstract>•New transgenic mice co-express G-CaMP6 and mCherry in ENS neurons.•Intestinal movement stabilization by suction enables imaging of ENS activity in living mice.•Spontaneous and evoked activity can be imaged at cellular and subcellular resolutions.
Most imaging studies of the enteric nervous system (ENS) that regulates the function of the gastrointestinal tract are so far performed using preparations isolated from animals, thus hindering the understanding of the ENS function in vivo. Here we report a method for imaging the ENS cellular network activity in living mice using a new transgenic mouse line that co-expresses G-CaMP6 and mCherry in the ENS combined with the suction-mediated stabilization of intestinal movements. With confocal or two-photon imaging, our method can visualize spontaneous and pharmacologically-evoked ENS network activity in living animals at cellular and subcellular resolutions, demonstrating the potential usefulness for studies of the ENS function in health and disease.</abstract><cop>Ireland</cop><pub>Elsevier B.V</pub><pmid>30790590</pmid><doi>10.1016/j.neures.2019.02.004</doi><tpages>8</tpages></addata></record> |
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| subjects | Animals Calcium - analysis Calcium - metabolism Confocal microscopy Enteric Nervous System - physiology Genetically-encoded calcium indicator Gut In vivo imaging Intestines Intravital Microscopy - methods Male Mice Mice, Transgenic Microscopy, Fluorescence - methods Molecular Imaging - methods Neurons - metabolism Neurons - physiology Serotonin Serotonin - pharmacology Transgenic mice Two-photon microscopy |
| title | Confocal and multiphoton calcium imaging of the enteric nervous system in anesthetized mice |
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