Protein kinase D isoforms are expressed in rat and mouse primary sensory neurons and are activated by agonists of protease-activated receptor 2

Amadesi, S., Grant, A. D., Cottrell, G. ORCID: https://orcid.org/0000-0001-9098-7627, Vaksmam, N., Poole, D. P., Rozengurt, E. and Bunnett, N. W. (2009) Protein kinase D isoforms are expressed in rat and mouse primary sensory neurons and are activated by agonists of protease-activated receptor 2. Journal of Comparative Neurology, 516 (2). pp. 141-156. ISSN 1096-9861 doi: 10.1002/cne.22104

Abstract/Summary

Serine proteases generated during injury and inflammation cleave protease-activated receptor 2 (PAR(2)) on primary sensory neurons to induce neurogenic inflammation and hyperalgesia. Hyperalgesia requires sensitization of transient receptor potential vanilloid (TRPV) ion channels by mechanisms involving phospholipase C and protein kinase C (PKC). The protein kinase D (PKD) serine/threonine kinases are activated by diacylglycerol and PKCs and can phosphorylate TRPV1. Thus, PKDs may participate in novel signal transduction pathways triggered by serine proteases during inflammation and pain. However, it is not known whether PAR(2) activates PKD, and the expression of PKD isoforms by nociceptive neurons is poorly characterized. By using HEK293 cells transfected with PKDs, we found that PAR(2) stimulation promoted plasma membrane translocation and phosphorylation of PKD1, PKD2, and PKD3, indicating activation. This effect was partially dependent on PKCepsilon. By immunofluorescence and confocal microscopy, with antibodies against PKD1/PKD2 and PKD3 and neuronal markers, we found that PKDs were expressed in rat and mouse dorsal root ganglia (DRG) neurons, including nociceptive neurons that expressed TRPV1, PAR(2), and neuropeptides. PAR(2) agonist induced phosphorylation of PKD in cultured DRG neurons, indicating PKD activation. Intraplantar injection of PAR(2) agonist also caused phosphorylation of PKD in neurons of lumbar DRG, confirming activation in vivo. Thus, PKD1, PKD2, and PKD3 are expressed in primary sensory neurons that mediate neurogenic inflammation and pain transmission, and PAR(2) agonists activate PKDs in HEK293 cells and DRG neurons in culture and in intact animals. PKD may be a novel component of a signal transduction pathway for protease-induced activation of nociceptive neurons and an important new target for antiinflammatory and analgesic therapies.

Additional Information	Full text freely available at PUBMED - see link at "Related URLs"
Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/30260
Item Type	Article
Refereed	Yes
Divisions	No Reading authors. Back catalogue items Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy > Division of Pharmacology
Uncontrolled Keywords	Animals Cell Line Cell Membrane/physiology Cells, Cultured Enzyme Activation Ganglia, Spinal/metabolism Humans Isoenzymes/metabolism Male Mice Mice, Inbred C57BL Neuropeptides/metabolism Nociceptors/*metabolism Protein Kinase C/*metabolism Protein Kinases/metabolism Rats Rats, Sprague-Dawley Receptor, PAR-2/*agonists/metabolism Sensory Receptor Cells/*metabolism TRPV Cation Channels/metabolism
Additional Information	Full text freely available at PUBMED - see link at "Related URLs"
Publisher	Wiley
Download/View statistics	View download statistics for this item