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.
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. 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.
authors.affiliation: Laboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, Antwerp, Belgium.
authors.forename: Roeland
authors.initials: R
authors.lastname: Buckinx
authors.forename: Luc
authors.initials: L
authors.lastname: Van Nassauw
authors.forename: Leela R
authors.initials: LR
authors.lastname: Avula
authors.forename: Katrien
authors.initials: K
authors.lastname: Alpaerts
authors.forename: Dirk
authors.initials: D
authors.lastname: Adriaensen
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Timmermans
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D017615:Enteric Nervous System / Q000737:chemistry* / Q000378:metabolism; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050916:TRPV Cation Channels / Q000032:analysis* / Q000235:genetics / Q000276:immunology*
chemical_list: D000906:Antibodies; D000939:Epitopes; D012333:RNA, Messenger; D050916:TRPV Cation Channels; C493921:TRPV1 protein, mouse
keywords: TRPV1; enteric nervous system; epitope; gastrointestinal tract