Transient Receptor Potential Vanilloid Type 1 Channel (TRPV1) Immunolocalization in the Murine Enteric Nervous System Is Affected by the Targeted C-terminal Epitope of the Applied Antibody
The expression of transient receptor potential vanilloid type 1 channel (TRPV1) in the enteric nervous system is still the subject of debate. Although a number of studies have reported that TRPV1 is limited to extrinsic afferent fibers, other studies argue for an intrinsic expression of TRPV1. In th...
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Veröffentlicht in: | The journal of histochemistry and cytochemistry 2013-06, Vol.61 (6), p.421-432 |
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container_title | The journal of histochemistry and cytochemistry |
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creator | Buckinx, Roeland Van Nassauw, Luc Avula, Leela R. Alpaerts, Katrien Adriaensen, Dirk Timmermans, Jean-Pierre |
description | The expression of transient receptor potential vanilloid type 1 channel (TRPV1) in the enteric nervous system is still the subject of debate. Although a number of studies have reported that TRPV1 is limited to extrinsic afferent fibers, other studies argue for an intrinsic expression of TRPV1. In the present study, reverse transcriptase PCR was employed to establish the expression of TRPV1 mRNA throughout the gastrointestinal tract. Using two antibodies directed against different epitopes of TRPV1, we were able to show at the protein level that the observed distribution pattern of TRPV1 is dependent on the antibody used in the immunohistochemical staining. A first antibody indeed mainly stained neuronal fibers, whereas a second antibody exclusively stained perikarya of enteric neurons throughout the mouse gastrointestinal tract. We argue that these different distribution patterns are due to the antibodies discriminating between different modulated forms of TRPV1 that influence the recognition of the targeted immunogen and as such distinguish intracellular from plasmalemmal forms of TRPV1. Our study is the first to directly compare these two antibodies within the same species and in identical conditions. Our observations underline that detailed knowledge of the epitope that is recognized by the antibodies employed in immunohistochemical procedures is a prerequisite for correctly interpreting experimental results. |
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Although a number of studies have reported that TRPV1 is limited to extrinsic afferent fibers, other studies argue for an intrinsic expression of TRPV1. In the present study, reverse transcriptase PCR was employed to establish the expression of TRPV1 mRNA throughout the gastrointestinal tract. Using two antibodies directed against different epitopes of TRPV1, we were able to show at the protein level that the observed distribution pattern of TRPV1 is dependent on the antibody used in the immunohistochemical staining. A first antibody indeed mainly stained neuronal fibers, whereas a second antibody exclusively stained perikarya of enteric neurons throughout the mouse gastrointestinal tract. We argue that these different distribution patterns are due to the antibodies discriminating between different modulated forms of TRPV1 that influence the recognition of the targeted immunogen and as such distinguish intracellular from plasmalemmal forms of TRPV1. Our study is the first to directly compare these two antibodies within the same species and in identical conditions. Our observations underline that detailed knowledge of the epitope that is recognized by the antibodies employed in immunohistochemical procedures is a prerequisite for correctly interpreting experimental results.</description><identifier>ISSN: 0022-1554</identifier><identifier>EISSN: 1551-5044</identifier><identifier>DOI: 10.1369/0022155413484764</identifier><identifier>PMID: 23482327</identifier><language>eng</language><publisher>Los Angeles, CA: SAGE Publications</publisher><subject>Animals ; Antibodies - immunology ; Antigen-Antibody Reactions ; Enteric Nervous System - chemistry ; Enteric Nervous System - metabolism ; Epitopes - chemistry ; Epitopes - immunology ; Immunohistochemistry ; Mice ; Mice, Inbred C57BL ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger - analysis ; RNA, Messenger - genetics ; TRPV Cation Channels - analysis ; TRPV Cation Channels - genetics ; TRPV Cation Channels - immunology</subject><ispartof>The journal of histochemistry and cytochemistry, 2013-06, Vol.61 (6), p.421-432</ispartof><rights>The Author(s) 2013</rights><rights>The Author(s) 2013 2013 The Histochemical Society</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c533t-65f8734d898c78f8786d6423e89373e87aef337c91d98ac6a6cd8c6fae1b7c603</citedby><cites>FETCH-LOGICAL-c533t-65f8734d898c78f8786d6423e89373e87aef337c91d98ac6a6cd8c6fae1b7c603</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://journals.sagepub.com/doi/pdf/10.1369/0022155413484764$$EPDF$$P50$$Gsage$$H</linktopdf><linktohtml>$$Uhttps://journals.sagepub.com/doi/10.1369/0022155413484764$$EHTML$$P50$$Gsage$$H</linktohtml><link.rule.ids>230,314,776,780,881,21799,27903,27904,43600,43601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23482327$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Buckinx, Roeland</creatorcontrib><creatorcontrib>Van Nassauw, Luc</creatorcontrib><creatorcontrib>Avula, Leela R.</creatorcontrib><creatorcontrib>Alpaerts, Katrien</creatorcontrib><creatorcontrib>Adriaensen, Dirk</creatorcontrib><creatorcontrib>Timmermans, Jean-Pierre</creatorcontrib><title>Transient Receptor Potential Vanilloid Type 1 Channel (TRPV1) Immunolocalization in the Murine Enteric Nervous System Is Affected by the Targeted C-terminal Epitope of the Applied Antibody</title><title>The journal of histochemistry and cytochemistry</title><addtitle>J Histochem Cytochem</addtitle><description>The expression of transient receptor potential vanilloid type 1 channel (TRPV1) in the enteric nervous system is still the subject of debate. Although a number of studies have reported that TRPV1 is limited to extrinsic afferent fibers, other studies argue for an intrinsic expression of TRPV1. In the present study, reverse transcriptase PCR was employed to establish the expression of TRPV1 mRNA throughout the gastrointestinal tract. Using two antibodies directed against different epitopes of TRPV1, we were able to show at the protein level that the observed distribution pattern of TRPV1 is dependent on the antibody used in the immunohistochemical staining. A first antibody indeed mainly stained neuronal fibers, whereas a second antibody exclusively stained perikarya of enteric neurons throughout the mouse gastrointestinal tract. We argue that these different distribution patterns are due to the antibodies discriminating between different modulated forms of TRPV1 that influence the recognition of the targeted immunogen and as such distinguish intracellular from plasmalemmal forms of TRPV1. Our study is the first to directly compare these two antibodies within the same species and in identical conditions. Our observations underline that detailed knowledge of the epitope that is recognized by the antibodies employed in immunohistochemical procedures is a prerequisite for correctly interpreting experimental results.</description><subject>Animals</subject><subject>Antibodies - immunology</subject><subject>Antigen-Antibody Reactions</subject><subject>Enteric Nervous System - chemistry</subject><subject>Enteric Nervous System - metabolism</subject><subject>Epitopes - chemistry</subject><subject>Epitopes - immunology</subject><subject>Immunohistochemistry</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Messenger - analysis</subject><subject>RNA, Messenger - genetics</subject><subject>TRPV Cation Channels - analysis</subject><subject>TRPV Cation Channels - genetics</subject><subject>TRPV Cation Channels - immunology</subject><issn>0022-1554</issn><issn>1551-5044</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFksFuEzEQhlcIREPhzgn5WA4L9tprey9IURQgUoGqhF5Xjnc2ceW1t7a3Ung2Hg6nKRUgIS62R_PNP-OZKYqXBL8hlDdvMa4qUteMUCaZ4OxRMcsmKWvM2ONidnCXB_9J8SzGa4wJY7V8WpxUma9oJWbFj3VQLhpwCV2ChjH5gC58yrZRFl0pZ6z1pkPr_QiIoMVOOQcWna0vL67Ia7Qahsl567Wy5rtKxjtkHEo7QJ-mYBygpUsQjEafIdz6KaKv-5hgQKuI5n0POkGHNvu7gLUKWzjYizKHDMbl_MvRJJ8T-_4OmY-jNZmY5-o2vts_L570ykZ4cX-fFt_eL9eLj-X5lw-rxfy81DWlqeR1LwVlnWykFjK_Je84qyjIhop8CgU9pUI3pGuk0lxx3UnNewVkIzTH9LR4d9Qdp80Anc7dCcq2YzCDCvvWK9P-6XFm1279bUsFqWnVZIGze4HgbyaIqR1M1GCtcpC70hLOMcNSYPF_lNacNlWeckbxEdXBxxigf6iI4PawH-3f-5FDXv3-k4eAXwuRgfIIRLWF9tpPIc8h_lvwJ1qtxTg</recordid><startdate>20130601</startdate><enddate>20130601</enddate><creator>Buckinx, Roeland</creator><creator>Van Nassauw, Luc</creator><creator>Avula, Leela R.</creator><creator>Alpaerts, Katrien</creator><creator>Adriaensen, Dirk</creator><creator>Timmermans, Jean-Pierre</creator><general>SAGE Publications</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>7TK</scope><scope>5PM</scope></search><sort><creationdate>20130601</creationdate><title>Transient Receptor Potential Vanilloid Type 1 Channel (TRPV1) Immunolocalization in the Murine Enteric Nervous System Is Affected by the Targeted C-terminal Epitope of the Applied Antibody</title><author>Buckinx, Roeland ; Van Nassauw, Luc ; Avula, Leela R. ; Alpaerts, Katrien ; Adriaensen, Dirk ; Timmermans, Jean-Pierre</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c533t-65f8734d898c78f8786d6423e89373e87aef337c91d98ac6a6cd8c6fae1b7c603</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>Antibodies - immunology</topic><topic>Antigen-Antibody Reactions</topic><topic>Enteric Nervous System - chemistry</topic><topic>Enteric Nervous System - metabolism</topic><topic>Epitopes - chemistry</topic><topic>Epitopes - immunology</topic><topic>Immunohistochemistry</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Messenger - analysis</topic><topic>RNA, Messenger - genetics</topic><topic>TRPV Cation Channels - analysis</topic><topic>TRPV Cation Channels - genetics</topic><topic>TRPV Cation Channels - immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Buckinx, Roeland</creatorcontrib><creatorcontrib>Van Nassauw, Luc</creatorcontrib><creatorcontrib>Avula, Leela R.</creatorcontrib><creatorcontrib>Alpaerts, Katrien</creatorcontrib><creatorcontrib>Adriaensen, Dirk</creatorcontrib><creatorcontrib>Timmermans, Jean-Pierre</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>Neurosciences Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The journal of histochemistry and cytochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Buckinx, Roeland</au><au>Van Nassauw, Luc</au><au>Avula, Leela R.</au><au>Alpaerts, Katrien</au><au>Adriaensen, Dirk</au><au>Timmermans, Jean-Pierre</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transient Receptor Potential Vanilloid Type 1 Channel (TRPV1) Immunolocalization in the Murine Enteric Nervous System Is Affected by the Targeted C-terminal Epitope of the Applied Antibody</atitle><jtitle>The journal of histochemistry and cytochemistry</jtitle><addtitle>J Histochem Cytochem</addtitle><date>2013-06-01</date><risdate>2013</risdate><volume>61</volume><issue>6</issue><spage>421</spage><epage>432</epage><pages>421-432</pages><issn>0022-1554</issn><eissn>1551-5044</eissn><abstract>The expression of transient receptor potential vanilloid type 1 channel (TRPV1) in the enteric nervous system is still the subject of debate. Although a number of studies have reported that TRPV1 is limited to extrinsic afferent fibers, other studies argue for an intrinsic expression of TRPV1. In the present study, reverse transcriptase PCR was employed to establish the expression of TRPV1 mRNA throughout the gastrointestinal tract. Using two antibodies directed against different epitopes of TRPV1, we were able to show at the protein level that the observed distribution pattern of TRPV1 is dependent on the antibody used in the immunohistochemical staining. A first antibody indeed mainly stained neuronal fibers, whereas a second antibody exclusively stained perikarya of enteric neurons throughout the mouse gastrointestinal tract. We argue that these different distribution patterns are due to the antibodies discriminating between different modulated forms of TRPV1 that influence the recognition of the targeted immunogen and as such distinguish intracellular from plasmalemmal forms of TRPV1. 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subjects | Animals Antibodies - immunology Antigen-Antibody Reactions Enteric Nervous System - chemistry Enteric Nervous System - metabolism Epitopes - chemistry Epitopes - immunology Immunohistochemistry Mice Mice, Inbred C57BL Rats Rats, Wistar Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - analysis RNA, Messenger - genetics TRPV Cation Channels - analysis TRPV Cation Channels - genetics TRPV Cation Channels - immunology |
title | Transient Receptor Potential Vanilloid Type 1 Channel (TRPV1) Immunolocalization in the Murine Enteric Nervous System Is Affected by the Targeted C-terminal Epitope of the Applied Antibody |
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