Disparate changes in the expression of transient receptor potential vanilloid type 1 receptor mRNA and protein in dorsal root ganglion neurons following local capsaicin treatment of the sciatic nerve in the rat

Abstract In situ hybridization, quantitative reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blot analysis were applied to study the changes in expression of the major nociceptive ion channel transient receptor potential vanilloid type 1 receptor (TRPV1) a...

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Published in:	Neuroscience Vol. 201; pp. 320 - 330
Main Authors:	Szigeti, C, Sántha, P, Körtvély, E, Nyári, T, Horváth, V.J, Deák, É, Dux, M, Gulya, K, Jancsó, G
Format:	Journal Article
Language:	English
Published:	Amsterdam Elsevier Ltd 10-01-2012 Elsevier
Subjects:	Analysis of Variance Animals Biological and medical sciences Capsaicin Capsaicin - adverse effects Capsaicin receptors Cell Count Cell survival Disease Models, Animal Dorsal root ganglia Fundamental and applied biological sciences. Psychology Ganglia, Spinal - pathology Gene expression Gene Expression Regulation - drug effects Immunohistochemistry in situ hybridization Inflammation Injuries Ion channels Male nerve transection Nerves Nervous system Neurology Neurons Neurons - drug effects Neurons - metabolism pain Pain perception perineural Polymerase chain reaction Rats Rats, Wistar Reverse transcription RNA, Messenger - metabolism Sciatic nerve Sciatic Neuropathy - chemically induced Sciatic Neuropathy - etiology Sciatic Neuropathy - pathology Sensory System Agents - adverse effects Statistical analysis Time Factors transient receptor potential proteins TRPV Cation Channels - genetics TRPV Cation Channels - metabolism TRPV1 mRNA Vertebrates: nervous system and sense organs Western blotting cross-sectional area gray value TRPV1 mRNA nerve transection CSA Tris-buffered saline nerve growth factor RT-PCR capsaicin beta-2-microglobulin transient receptor potential vanilloid type 1 receptor receiver operating characteristic PBS pain TBS ROD ROC TRPV1 NGF B2-MG GV phosphate buffered saline dorsal root ganglion DRG reverse transcription polymerase chain reaction relative optical density perineural in situ hybridization Molecular hybridization Spinal cord Rat Capsaicin Rodentia Central nervous system Protein Vertebrata TRPV1 vanilloid receptor Mammalia Pain Neuron Animal Spinal ganglion Vanilloid receptor Sciatic nerve Biological receptor
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Summary:	Abstract In situ hybridization, quantitative reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blot analysis were applied to study the changes in expression of the major nociceptive ion channel transient receptor potential vanilloid type 1 receptor (TRPV1) after the perineural application of capsaicin or nerve transection. In control rats, quantitative morphometric and statistical analyses of TRPV1 protein and mRNA expression in L5 dorsal root ganglion cells revealed distinct populations of small (type C) and small-to-medium (type B) neurons, which showed very high and moderate levels of TRPV1, whereas larger (type A) neurons mostly did not express this receptor. After either transection or capsaicin treatment of the sciatic nerve, immunohistochemistry and Western blotting demonstrated a massive (up to 80%) decrease in the proportion of TRPV1-immunoreactive neurons and TRPV1 protein at all postoperative survival times. In situ hybridization indicated marked decreases (up to 85%) in the proportion of neurons that expressed TRPV1 mRNA after sciatic nerve transection. In contrast, although perineural treatment with capsaicin resulted in similar substantial decreases in the proportions of type B and C neurons of the L5 dorsal root ganglia 3 days postoperatively, a clear-cut tendency to recovery was observed thereafter. Hence, the proportions of both type B and C neurons expressing TRPV1 mRNA reached up to 70% of the control levels at 30 days postoperatively. In accord with these findings, quantitative RT-PCR revealed a marked and significant recovery in TRPV1 mRNA after perineural capsaicin but not after nerve transection. These observations suggest the involvement of distinct cellular mechanisms in the regulation of the TRPV1 mRNA expression of damaged neurons, specifically triggered by the nature of the injury. The present findings imply that the antinociceptive and anti-inflammatory effects of perineurally applied capsaicin involve distinct changes in neuronal TRPV1 mRNA expression and long-lasting alterations in (post)translational regulation.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1
ISSN:	0306-4522 1873-7544
DOI:	10.1016/j.neuroscience.2011.10.058