title: Pain modulators regulate the dynamics of PKA-RII phosphorylation in subgroups of sensory neurons.
abstract: Knowledge about the molecular structure of protein kinase A (PKA) isoforms is substantial. In contrast, the dynamics of PKA isoform activity in living primary cells has not been investigated in detail. Using a high content screening microscopy approach, we identified the RIIβ subunit of PKA-II to be predominantly expressed in a subgroup of sensory neurons. The RIIβ-positive subgroup included most neurons expressing nociceptive markers (TRPV1, NaV1.8, CGRP, IB4) and responded to pain-eliciting capsaicin with calcium influx. Isoform-specific PKA reporters showed in sensory-neuron-derived F11 cells that the inflammatory mediator PGE₂ specifically activated PKA-II but not PKA-I. Accordingly, pain-sensitizing inflammatory mediators and activators of PKA increased the phosphorylation of RII subunits (pRII) in subgroups of primary sensory neurons. Detailed analyses revealed basal pRII to be regulated by the phosphatase PP2A. Increase of pRII was followed by phosphorylation of CREB in a PKA-dependent manner. Thus, we propose RII phosphorylation to represent an isoform-specific readout for endogenous PKA-II activity in vivo, suggest RIIβ as a novel nociceptive subgroup marker, and extend the current model of PKA-II activation by introducing a PP2A-dependent basal state.
authors.affiliation: University Hospital Cologne, Department of Anesthesiology and Intensive Care Medicine, Experimental Anesthesiology and Pain Research, Robert Koch Str. 10, 50931 Cologne, Germany.
authors.forename: Joerg
authors.initials: J
authors.lastname: Isensee
authors.forename: Mandy
authors.initials: M
authors.lastname: Diskar
authors.forename: Steffen
authors.initials: S
authors.lastname: Waldherr
authors.forename: René
authors.initials: R
authors.lastname: Buschow
authors.forename: Jan
authors.initials: J
authors.lastname: Hasenauer
authors.forename: Anke
authors.initials: A
authors.lastname: Prinz
authors.forename: Frank
authors.initials: F
authors.lastname: Allgöwer
authors.forename: Friedrich W
authors.initials: FW
authors.lastname: Herberg
authors.forename: Tim
authors.initials: T
authors.lastname: Hucho
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015740:Calcitonin Gene-Related Peptide / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002211:Capsaicin / Q000494:pharmacology*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D054757:Cyclic AMP-Dependent Protein Kinase RIIbeta Subunit / Q000235:genetics / Q000378:metabolism; D054743:Cyclic AMP-Dependent Protein Kinase Type I / Q000235:genetics / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology; D005786:Gene Expression Regulation; D008297:Male; D062559:NAV1.8 Voltage-Gated Sodium Channel / Q000235:genetics / Q000378:metabolism; D059225:Nociception / Q000187:drug effects*; D010766:Phosphorylation; D061251:Primary Cell Culture; D054648:Protein Phosphatase 2 / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011984:Sensory Receptor Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015398:Signal Transduction; D050916:TRPV Cation Channels / Q000235:genetics / Q000378:metabolism
chemical_list: D015415:Biomarkers; D054757:Cyclic AMP-Dependent Protein Kinase RIIbeta Subunit; D062559:NAV1.8 Voltage-Gated Sodium Channel; C568319:Scn10a protein, rat; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat; D005576:Colforsin; D016572:Cyclosporine; D000242:Cyclic AMP; D054743:Cyclic AMP-Dependent Protein Kinase Type I; D054648:Protein Phosphatase 2; D015740:Calcitonin Gene-Related Peptide; D015232:Dinoprostone; D002211:Capsaicin; D002118:Calcium
keywords: Nociception; Protein kinase A; RII phosphorylation; Sensitization; cAMP response element-binding protein