Functional characterization of transient receptor potential channels in mouse urothelial cells
The bladder urothelium is currently believed to be a sensory structure, contributing to mechano- and chemosensation in the bladder. Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TR...
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| Veröffentlicht in: | American Journal of Physiology - Renal Physiology 2010-03, Vol.298 (3), p.F692-F701 |
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| container_title | American Journal of Physiology - Renal Physiology |
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| creator | Everaerts, Wouter Vriens, Joris Owsianik, Grzegorz Appendino, Giovanni Voets, Thomas De Ridder, Dirk Nilius, Bernd |
| description | The bladder urothelium is currently believed to be a sensory structure, contributing to mechano- and chemosensation in the bladder. Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TRP channel expression profile of urothelial cells is unclear. In this study, we have performed a systematic analysis of the molecular and functional expression of various TRP channels in mouse urothelium. Urothelial cells from control and trpv4-/- mice were isolated, cultured (12-48 h), and used for quantitative real-time PCR, immunocytochemistry, calcium imaging, and whole cell patch-clamp experiments. At the mRNA level, TRPV4, TRPV2, and TRPM7 were the most abundantly expressed TRP genes. Immunohistochemistry showed a clear expression of TRPV4 in the plasma membrane, whereas TRPV2 was more prominent in the cytoplasm. TRPM7 was detected in the plasma membrane as well as cytoplasmic vesicles. Calcium imaging and patch-clamp experiments using TRP channel agonists and antagonists provided evidence for the functional expression of TRPV4, TRPV2, and TRPM7 but not of TRPA1, TRPV1, and TRPM8. In conclusion, we have demonstrated functional expression of TRPV4, TRPV2, and TRPM7 in mouse urothelial cells. These channels may contribute to the (mechano)sensory function of the urothelial layer and represent potential targets for the treatment of bladder dysfunction. |
| doi_str_mv | 10.1152/ajprenal.00599.2009 |
| format | Article |
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Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TRP channel expression profile of urothelial cells is unclear. In this study, we have performed a systematic analysis of the molecular and functional expression of various TRP channels in mouse urothelium. Urothelial cells from control and trpv4-/- mice were isolated, cultured (12-48 h), and used for quantitative real-time PCR, immunocytochemistry, calcium imaging, and whole cell patch-clamp experiments. At the mRNA level, TRPV4, TRPV2, and TRPM7 were the most abundantly expressed TRP genes. Immunohistochemistry showed a clear expression of TRPV4 in the plasma membrane, whereas TRPV2 was more prominent in the cytoplasm. TRPM7 was detected in the plasma membrane as well as cytoplasmic vesicles. Calcium imaging and patch-clamp experiments using TRP channel agonists and antagonists provided evidence for the functional expression of TRPV4, TRPV2, and TRPM7 but not of TRPA1, TRPV1, and TRPM8. In conclusion, we have demonstrated functional expression of TRPV4, TRPV2, and TRPM7 in mouse urothelial cells. These channels may contribute to the (mechano)sensory function of the urothelial layer and represent potential targets for the treatment of bladder dysfunction.</description><identifier>ISSN: 1931-857X</identifier><identifier>ISSN: 0363-6127</identifier><identifier>EISSN: 1522-1466</identifier><identifier>DOI: 10.1152/ajprenal.00599.2009</identifier><identifier>PMID: 20015940</identifier><language>eng</language><publisher>United States: American Physiological Society</publisher><subject>Animals ; Bladder ; Calcium Channels - metabolism ; Calcium Signaling - drug effects ; Cells ; Cells, Cultured ; Fluorescent Antibody Technique ; Gene expression ; Kidneys ; Membrane Potentials ; Membrane Transport Modulators - pharmacology ; Membranes ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Microscopy, Fluorescence ; Molecular biology ; Patch-Clamp Techniques ; Polymerase Chain Reaction ; RNA, Messenger - metabolism ; Rodents ; T cell receptors ; Time Factors ; Transient Receptor Potential Channels - deficiency ; Transient Receptor Potential Channels - drug effects ; Transient Receptor Potential Channels - genetics ; Transient Receptor Potential Channels - metabolism ; TRPM Cation Channels - metabolism ; TRPV Cation Channels - metabolism ; Urinary Bladder - cytology ; Urinary Bladder - drug effects ; Urinary Bladder - metabolism ; Urothelium - metabolism</subject><ispartof>American Journal of Physiology - Renal Physiology, 2010-03, Vol.298 (3), p.F692-F701</ispartof><rights>Copyright American Physiological Society Mar 2010</rights><rights>Copyright © 2010 the American Physiological Society</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c497t-4e11a2d1c65bc0be399e924a8f74c8a0070c3c2eec24c40d7477d16ae83b2afc3</citedby><cites>FETCH-LOGICAL-c497t-4e11a2d1c65bc0be399e924a8f74c8a0070c3c2eec24c40d7477d16ae83b2afc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,778,782,883,3028,27911,27912</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20015940$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Everaerts, Wouter</creatorcontrib><creatorcontrib>Vriens, Joris</creatorcontrib><creatorcontrib>Owsianik, Grzegorz</creatorcontrib><creatorcontrib>Appendino, Giovanni</creatorcontrib><creatorcontrib>Voets, Thomas</creatorcontrib><creatorcontrib>De Ridder, Dirk</creatorcontrib><creatorcontrib>Nilius, Bernd</creatorcontrib><title>Functional characterization of transient receptor potential channels in mouse urothelial cells</title><title>American Journal of Physiology - Renal Physiology</title><addtitle>Am J Physiol Renal Physiol</addtitle><description>The bladder urothelium is currently believed to be a sensory structure, contributing to mechano- and chemosensation in the bladder. Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TRP channel expression profile of urothelial cells is unclear. In this study, we have performed a systematic analysis of the molecular and functional expression of various TRP channels in mouse urothelium. Urothelial cells from control and trpv4-/- mice were isolated, cultured (12-48 h), and used for quantitative real-time PCR, immunocytochemistry, calcium imaging, and whole cell patch-clamp experiments. At the mRNA level, TRPV4, TRPV2, and TRPM7 were the most abundantly expressed TRP genes. Immunohistochemistry showed a clear expression of TRPV4 in the plasma membrane, whereas TRPV2 was more prominent in the cytoplasm. TRPM7 was detected in the plasma membrane as well as cytoplasmic vesicles. Calcium imaging and patch-clamp experiments using TRP channel agonists and antagonists provided evidence for the functional expression of TRPV4, TRPV2, and TRPM7 but not of TRPA1, TRPV1, and TRPM8. In conclusion, we have demonstrated functional expression of TRPV4, TRPV2, and TRPM7 in mouse urothelial cells. These channels may contribute to the (mechano)sensory function of the urothelial layer and represent potential targets for the treatment of bladder dysfunction.</description><subject>Animals</subject><subject>Bladder</subject><subject>Calcium Channels - metabolism</subject><subject>Calcium Signaling - drug effects</subject><subject>Cells</subject><subject>Cells, Cultured</subject><subject>Fluorescent Antibody Technique</subject><subject>Gene expression</subject><subject>Kidneys</subject><subject>Membrane Potentials</subject><subject>Membrane Transport Modulators - pharmacology</subject><subject>Membranes</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Mice, Knockout</subject><subject>Microscopy, Fluorescence</subject><subject>Molecular biology</subject><subject>Patch-Clamp Techniques</subject><subject>Polymerase Chain Reaction</subject><subject>RNA, Messenger - metabolism</subject><subject>Rodents</subject><subject>T cell receptors</subject><subject>Time Factors</subject><subject>Transient Receptor Potential Channels - deficiency</subject><subject>Transient Receptor Potential Channels - drug effects</subject><subject>Transient Receptor Potential Channels - genetics</subject><subject>Transient Receptor Potential Channels - metabolism</subject><subject>TRPM Cation Channels - metabolism</subject><subject>TRPV Cation Channels - metabolism</subject><subject>Urinary Bladder - cytology</subject><subject>Urinary Bladder - drug effects</subject><subject>Urinary Bladder - metabolism</subject><subject>Urothelium - metabolism</subject><issn>1931-857X</issn><issn>0363-6127</issn><issn>1522-1466</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdUctKxDAUDaL4_gJBihtXHfNq02wEEV8guFFwZchkbp0MnaQmqaBfb-qoqKsk9zy4JwehA4InhFT0RC_6AE53E4wrKScUY7mGtjNCS8Lrej3fJSNlU4nHLbQT4wJjTAglm2grc0klOd5GT5eDM8n67FOYuQ7aJAj2XY-jwrdFCtpFCy4VAQz0yYei9ym_7UrgHHSxsK5Y-iFCMQSf5tB9gtB1cQ9ttLqLsP917qKHy4v78-vy9u7q5vzstjRcilRyIETTGTF1NTV4CkxKkJTrphXcNBpjgQ0zFMBQbjieCS7EjNQaGjalujVsF52ufPthuoSZyQsG3ak-2KUOb8prq_4izs7Vs39VtGFNJUU2OP4yCP5lgJjU0sYxgnaQkynBWC1oTXlmHv1jLvwQ8v9FRRkmvKr5aMdWJBN8jAHan1UIVmN76rs99dmeGtvLqsPfKX4033WxD9xJm3c</recordid><startdate>20100301</startdate><enddate>20100301</enddate><creator>Everaerts, Wouter</creator><creator>Vriens, Joris</creator><creator>Owsianik, Grzegorz</creator><creator>Appendino, Giovanni</creator><creator>Voets, Thomas</creator><creator>De Ridder, Dirk</creator><creator>Nilius, Bernd</creator><general>American Physiological Society</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><scope>5PM</scope></search><sort><creationdate>20100301</creationdate><title>Functional characterization of transient receptor potential channels in mouse urothelial cells</title><author>Everaerts, Wouter ; Vriens, Joris ; Owsianik, Grzegorz ; Appendino, Giovanni ; Voets, Thomas ; De Ridder, Dirk ; Nilius, Bernd</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c497t-4e11a2d1c65bc0be399e924a8f74c8a0070c3c2eec24c40d7477d16ae83b2afc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Animals</topic><topic>Bladder</topic><topic>Calcium Channels - metabolism</topic><topic>Calcium Signaling - drug effects</topic><topic>Cells</topic><topic>Cells, Cultured</topic><topic>Fluorescent Antibody Technique</topic><topic>Gene expression</topic><topic>Kidneys</topic><topic>Membrane Potentials</topic><topic>Membrane Transport Modulators - pharmacology</topic><topic>Membranes</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Mice, Knockout</topic><topic>Microscopy, Fluorescence</topic><topic>Molecular biology</topic><topic>Patch-Clamp Techniques</topic><topic>Polymerase Chain Reaction</topic><topic>RNA, Messenger - metabolism</topic><topic>Rodents</topic><topic>T cell receptors</topic><topic>Time Factors</topic><topic>Transient Receptor Potential Channels - deficiency</topic><topic>Transient Receptor Potential Channels - drug effects</topic><topic>Transient Receptor Potential Channels - genetics</topic><topic>Transient Receptor Potential Channels - metabolism</topic><topic>TRPM Cation Channels - metabolism</topic><topic>TRPV Cation Channels - metabolism</topic><topic>Urinary Bladder - cytology</topic><topic>Urinary Bladder - drug effects</topic><topic>Urinary Bladder - metabolism</topic><topic>Urothelium - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Everaerts, Wouter</creatorcontrib><creatorcontrib>Vriens, Joris</creatorcontrib><creatorcontrib>Owsianik, Grzegorz</creatorcontrib><creatorcontrib>Appendino, Giovanni</creatorcontrib><creatorcontrib>Voets, Thomas</creatorcontrib><creatorcontrib>De Ridder, Dirk</creatorcontrib><creatorcontrib>Nilius, Bernd</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><collection>PubMed Central (Full Participant titles)</collection><jtitle>American Journal of Physiology - Renal Physiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Everaerts, Wouter</au><au>Vriens, Joris</au><au>Owsianik, Grzegorz</au><au>Appendino, Giovanni</au><au>Voets, Thomas</au><au>De Ridder, Dirk</au><au>Nilius, Bernd</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Functional characterization of transient receptor potential channels in mouse urothelial cells</atitle><jtitle>American Journal of Physiology - Renal Physiology</jtitle><addtitle>Am J Physiol Renal Physiol</addtitle><date>2010-03-01</date><risdate>2010</risdate><volume>298</volume><issue>3</issue><spage>F692</spage><epage>F701</epage><pages>F692-F701</pages><issn>1931-857X</issn><issn>0363-6127</issn><eissn>1522-1466</eissn><abstract>The bladder urothelium is currently believed to be a sensory structure, contributing to mechano- and chemosensation in the bladder. Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TRP channel expression profile of urothelial cells is unclear. In this study, we have performed a systematic analysis of the molecular and functional expression of various TRP channels in mouse urothelium. Urothelial cells from control and trpv4-/- mice were isolated, cultured (12-48 h), and used for quantitative real-time PCR, immunocytochemistry, calcium imaging, and whole cell patch-clamp experiments. At the mRNA level, TRPV4, TRPV2, and TRPM7 were the most abundantly expressed TRP genes. Immunohistochemistry showed a clear expression of TRPV4 in the plasma membrane, whereas TRPV2 was more prominent in the cytoplasm. TRPM7 was detected in the plasma membrane as well as cytoplasmic vesicles. Calcium imaging and patch-clamp experiments using TRP channel agonists and antagonists provided evidence for the functional expression of TRPV4, TRPV2, and TRPM7 but not of TRPA1, TRPV1, and TRPM8. In conclusion, we have demonstrated functional expression of TRPV4, TRPV2, and TRPM7 in mouse urothelial cells. These channels may contribute to the (mechano)sensory function of the urothelial layer and represent potential targets for the treatment of bladder dysfunction.</abstract><cop>United States</cop><pub>American Physiological Society</pub><pmid>20015940</pmid><doi>10.1152/ajprenal.00599.2009</doi><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 1931-857X |
| ispartof | American Journal of Physiology - Renal Physiology, 2010-03, Vol.298 (3), p.F692-F701 |
| issn | 1931-857X 0363-6127 1522-1466 |
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| source | MEDLINE; American Physiological Society; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Animals Bladder Calcium Channels - metabolism Calcium Signaling - drug effects Cells Cells, Cultured Fluorescent Antibody Technique Gene expression Kidneys Membrane Potentials Membrane Transport Modulators - pharmacology Membranes Mice Mice, Inbred C57BL Mice, Knockout Microscopy, Fluorescence Molecular biology Patch-Clamp Techniques Polymerase Chain Reaction RNA, Messenger - metabolism Rodents T cell receptors Time Factors Transient Receptor Potential Channels - deficiency Transient Receptor Potential Channels - drug effects Transient Receptor Potential Channels - genetics Transient Receptor Potential Channels - metabolism TRPM Cation Channels - metabolism TRPV Cation Channels - metabolism Urinary Bladder - cytology Urinary Bladder - drug effects Urinary Bladder - metabolism Urothelium - metabolism |
| title | Functional characterization of transient receptor potential channels in mouse urothelial cells |
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