CaMKII binding to GluN2B at S1303 has no role in acute or inflammatory pain

• Published in: Brain research Vol. 1750; p. 147154
• Main Authors: Maduka, Uche P., White, Stephanie R., Joiner, Mei-Ling A., Hell, Johannes W., Hammond, Donna L.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-01-2021
• Subjects: Animals; Calcium Signaling - physiology; Calcium-Calmodulin-Dependent Protein Kinase Type 2 - metabolism; Calcium-Calmodulin-Dependent Protein Kinase Type 2 - physiology; Calmodulin - metabolism; CaMKII; Capsaicin; Capsaicin - metabolism; Female; GluN2B; Inflammation - metabolism; Male; Mice; Mice, Inbred C57BL; NMDA receptor; Nociception; Nociception - physiology; Pain - metabolism; Pain - physiopathology; Protein Binding; Receptors, N-Methyl-D-Aspartate - metabolism; Receptors, N-Methyl-D-Aspartate - physiology; mN; DMSO; Capsaicin; K; CaM; ANOVA; EF50; WT; KI; KN93; GluN2B; Nociception; GluN1; s.c; siRNA; S.E.M; CaMKII; CAP; Ca2; HET; Na+; NMDAR; MK801; NMDA receptor; PCR
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/j.brainres.2020.147154

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•Acute and inflammatory pain were tested in GluN2BKI mice impaired in CaMKII binding.•Loss of activity-driven CaMKII binding to GluN2B did not diminish acute pain.•Loss of activity-driven CaMKII binding to GluN2B did not obtund inflammatory pain.•KN93 inhibited formalin pain in GluN2BKI mice implicating other targets of Ca2+/CaM.•CaMKII binding to S1303 in GluN2B is not critical for inflammatory pain signaling.
Activation of Ca2+/calmodulin kinase II (CaMKII) and the N-Methyl D-aspartate receptor (NMDAR), particularly its GluN2B subunit, contribute to the central sensitization of nociceptive pathways and persistent pain. Using mutant mice wherein the activity-driven binding of CaMKII to S1303 in GluN2B is abrogated (GluN2BKI), this study investigated the importance of this interaction for acute and persistent inflammatory nociception. GluN2BKI, wild type and heterozygote mice did not differ in responses to acute noxious heat stimuli as measured with tail flick, paw flick, or hot plate assays, nor did they differ in their responses to mechanical stimulation with von Frey filaments. Surprisingly, the three genotypes exhibited similar spontaneous pain behaviors and hypersensitivity to heat or mechanical stimuli induced by intraplantar injection of capsaicin; however, GluN2BKI mice did not immediately attend to the paw. WT and GluN2BKI mice also did not differ in the nociceptive behaviors elicited by intraplantar injection of formalin, even though MK801 greatly reduced these behaviors in both genotypes concordant with NMDAR dependence. CaMKII binding to GluN2B at S1303 therefore does not appear to be critical for the development of inflammatory nociception. Finally, intrathecal KN93 reduced formalin-induced nociceptive behaviors in GluN2BKI mice. KN93 does not inhibit CaKMII, but rather binds Ca2+/calmodulin. It has multiple other targets including Ca2+-, Na+- and K+-channels, as well as various kinases. Therefore, the use of GluN2BKI mice provided genetic specificity in assessing the role of CaMKII in inflammatory pain signaling cascades. These results challenge current thinking on the involvement of the CaMKII-NMDAR interaction in inflammatory pain.