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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.
- Source :
-
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society [J Histochem Cytochem] 2013 Jun; Vol. 61 (6), pp. 421-32. Date of Electronic Publication: 2013 Mar 12. - Publication Year :
- 2013
-
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.
- Subjects :
- Animals
Antigen-Antibody Reactions
Enteric Nervous System metabolism
Epitopes chemistry
Immunohistochemistry
Mice
Mice, Inbred C57BL
RNA, Messenger analysis
RNA, Messenger genetics
Rats
Rats, Wistar
Reverse Transcriptase Polymerase Chain Reaction
TRPV Cation Channels genetics
Antibodies immunology
Enteric Nervous System chemistry
Epitopes immunology
TRPV Cation Channels analysis
TRPV Cation Channels immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1551-5044
- Volume :
- 61
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
- Publication Type :
- Academic Journal
- Accession number :
- 23482327
- Full Text :
- https://doi.org/10.1369/0022155413484764