title: Phosphorylated CaMKII post-synaptic binding to NR2B subunits in the anterior cingulate cortex mediates visceral pain in visceral hypersensitive rats.
abstract: The NR2B subunit of NMDA receptor in the anterior cingulate cortex (ACC) is up-regulated in viscerally hypersensitive (VH) rats induced by colonic anaphylaxis. It plays a critical role in modulation of ACC sensitization and visceral pain responses. Given the key role of calcium/calmodulin-dependent protein kinase II (CaMKII) in synaptic plasticity and behavior learning and memory, we hypothesize that phosphorylation of CaMKII binding to NR2B mediates visceral pain in VH states. We performed in vivo electroporation of CaMKII siRNA produced inhibition of colorectal distension-induced visceromotor response in the VH rats. The NR2B, CaMKII and P-CaMKII-Thr²⁸⁶ protein levels were increased in 180%, 220% and 304% fold in the post-synaptic density (PSD) fraction in VH rats separately. Western blotting following co-immunoprecipitation showed that P-CaMKII-Thr²⁸⁶ bound to NR2B in the PSD, which was increased to 267% of control in VH rats. Administration of CaMKII antagonist Antennapedia-CaMKIINtide suppressed visceromotor response in VH rats in parallel with decrease of NR2B levels and reduction of the NR2B-P-CaMKII-Thr²⁸⁶ protein complex in PSD. In conclusion, CaMKII is a critical signaling molecule in the ACC glutamatergic synaptic transmission and phosphorylation of CaMKII at Thr286, which binds to NR2B subunit at post-synaptic site, modulates visceral pain in viscerally hypersensitive state.
authors.affiliation: Neuroscience Laboratory, Department of Biology and Chemistry, the City University of Hong Kong, Kowloon, Hong Kong. yingli@cityu.edu.hk
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
authors.forename: Haiyan
authors.initials: H
authors.lastname: Liu
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Cao
authors.forename: Shengliang
authors.initials: S
authors.lastname: Chen
authors.forename: Bing
authors.initials: B
authors.lastname: Cao
authors.forename: Jin
authors.initials: J
authors.lastname: Liu
mesh_terms: D000707:Anaphylaxis / Q000150:complications; D000818:Animals; D015153:Blotting, Western; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism* / Q000502:physiology*; D002404:Catheterization; D003108:Colonic Diseases / Q000150:complications; D018274:Electroporation; D005822:Genetic Vectors; D018698:Glutamic Acid / Q000378:metabolism; D006179:Gyrus Cinguli / Q000378:metabolism* / Q000502:physiology*; D006930:Hyperalgesia / Q000503:physiopathology*; D047468:Immunoprecipitation; D008297:Male; D010455:Peptides; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism* / Q000502:physiology*; D013347:Subcellular Fractions / Q000378:metabolism; D013569:Synapses / Q000378:metabolism* / Q000502:physiology*; D013574:Synaptosomes / Q000378:metabolism; D059265:Visceral Pain / Q000503:physiopathology*
chemical_list: C121001:NR2B NMDA receptor; D010455:Peptides; D034741:RNA, Small Interfering; D016194:Receptors, N-Methyl-D-Aspartate; C520570:antennapedia-CaMKIINtide; D018698:Glutamic Acid; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2