title: Opioid withdrawal increases transient receptor potential vanilloid 1 activity in a protein kinase A-dependent manner.
abstract: Hyperalgesia is a cardinal symptom of opioid withdrawal. The transient receptor potential vanilloid 1 (TRPV1) is a ligand-gated ion channel expressed on sensory neurons responding to noxious heat, protons, and chemical stimuli such as capsaicin. TRPV1 can be inhibited via μ-opioid receptor (MOR)-mediated reduced activity of adenylyl cyclases (ACs) and decreased cyclic adenosine monophosphate (cAMP) levels. In contrast, opioid withdrawal following chronic activation of MOR uncovers AC superactivation and subsequent increases in cAMP and protein kinase A (PKA) activity. Here we investigated (1) whether an increase in cAMP during opioid withdrawal increases the activity of TRPV1 and (2) how opioid withdrawal modulates capsaicin-induced nocifensive behavior in rats. We applied whole-cell patch clamp, microfluorimetry, cAMP assays, radioligand binding, site-directed mutagenesis, and behavioral experiments. Opioid withdrawal significantly increased cAMP levels and capsaicin-induced TRPV1 activity in both transfected human embryonic kidney 293 cells and dissociated dorsal root ganglion (DRG) neurons. Inhibition of AC and PKA, as well as mutations of the PKA phosphorylation sites threonine 144 and serine 774, prevented the enhanced TRPV1 activity. Finally, capsaicin-induced nocifensive behavior was increased during opioid withdrawal in vivo. In summary, our results demonstrate an increased activity of TRPV1 in DRG neurons as a new mechanism contributing to opioid withdrawal-induced hyperalgesia.
authors: Charité - Universitätsmedizin Berlin, Klinik für Anaesthesiologie und operative Intensivmedizin, Campus Benjamin Franklin, Hindenburgdamm 30, Berlin 12203, Germany Universitätsklinikum Hamburg - Eppendorf, Klinik für Anästhesiologie, Hamburg 20251, Germany.
authors: Viola
authors: V
authors: Spahn
authors: Oliver
authors: O
authors: Fischer
authors: Jeannette
authors: J
authors: Endres-Becker
authors: Michael
authors: M
authors: Schäfer
authors: Christoph
authors: C
authors: Stein
authors: Christian
authors: C
authors: Zöllner
mesh_terms: D000701:Analgesics, Opioid / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002211:Capsaicin / Q000009:adverse effects; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004195:Disease Models, Animal; D004224:Diterpenes / Q000493:pharmacokinetics; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)- / Q000493:pharmacokinetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005283:Fentanyl / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D006930:Hyperalgesia / Q000139:chemically induced / Q000188:drug therapy / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D009020:Morphine / Q000494:pharmacology; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding / Q000187:drug effects; D051381:Rats; D017450:Receptors, Opioid, mu / Q000378:metabolism; D011984:Sensory Receptor Cells / Q000187:drug effects* / Q000378:metabolism; D013375:Substance Withdrawal Syndrome / Q000378:metabolism; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism*; D014316:Tritium / Q000493:pharmacokinetics
chemical_list: D000701:Analgesics, Opioid; D004224:Diterpenes; D004791:Enzyme Inhibitors; D017450:Receptors, Opioid, mu; D050916:TRPV Cation Channels; C494482:TRPV1 protein, human; D014316:Tritium; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D009020:Morphine; C024353:resiniferatoxin; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D002211:Capsaicin; D002118:Calcium; D005283:Fentanyl
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