Activated PKA and PKC, but not CaMKIIalpha, are required for AMPA/Kainate-mediated pain behavior in the thermal stimulus model.
Secondary mechanical allodynia resulting from a thermal stimulus (52.5 degrees C for 45s) is blocked by intrathecal (i.t.) pretreatment with calcium-permeable AMPA/KA receptor antagonists, but not NMDA receptor antagonists. Spinal sensitization is presumed to underlie thermal stimulus-evoked secondary mechanical allodynia. We investigated whether this spinal sensitization involves activation and phosphorylation of calcium-dependent protein kinases (PKA, PKC and CaMKIIalpha), and examined if the noxious stimulus increases phosphorylated AMPA GLUR1 (pGLUR1 Ser-845 and pGLUR1 Ser-831). Secondary mechanical allodynia after thermal stimulation was not altered by i.t. pretreatment with control vehicles (saline or 5% DMSO). Comparable allodynia was observed after pretreatment with a selective CaMKIIalpha inhibitor (17 and 34nmol KN-93). In marked contrast, pretreatment with either a PKA (10nmol H89) or PKC (30nmol chelerythrine) inhibitor blocked allodynia. Western immunoblot analyses supported behavioral findings and revealed a thermal stimulus-evoked increase in spinal phosphorylated PKA and PKC, but not CaMKIIalpha. There was no increase in any of the total protein kinases. Although thermal stimulation did not change either pGLUR1 Ser-845 or pGLUR1 Ser-831, it was associated with an increase in cytosolic total GLUR1. Pretreatment with a selective calcium-permeable AMPA/KA receptor antagonist (5nmol joro spider toxin), but not an NMDA receptor antagonist (25nmol d-2-amino-5-phosphonovalerate, AP-5), blocked thermal stimulus-evoked increases in phosphorylated PKA and PKC, in addition to increased cytosolic GLUR1. These findings indicate that spinal sensitization in the thermal stimulus model does not involve CaMKIIalpha activation or AMPA GLUR1 receptor phosphorylation, and differs from that occurring in NMDAr-dependent pain states.
Jones
Toni L
TL
The University of California-San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0818, USA.
Sorkin
Linda S
LS
D015763:2-Amino-5-phosphonovalerate / Q000494:pharmacology; D000818:Animals; D019540:Area Under Curve; D015153:Blotting, Western / Q000379:methods; D002211:Capsaicin / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D003952:Diagnostic Imaging / Q000379:methods; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000627:therapeutic use; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D006358:Hot Temperature; D006930:Hyperalgesia / Q000188:drug therapy / Q000378:metabolism* / Q000503:physiopathology; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D010146:Pain / Q000188:drug therapy / Q000378:metabolism*; D010147:Pain Measurement / Q000379:methods; D010766:Phosphorylation / Q000187:drug effects; D010812:Physical Stimulation / Q000379:methods; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D018091:Receptors, AMPA / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D013111:Spider Venoms / Q000494:pharmacology; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism*; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000378:metabolism*
D015763:2-Amino-5-phosphonovalerate; D002211:Capsaicin; D017868:Cyclic AMP-Dependent Protein Kinases; D004791:Enzyme Inhibitors; D018691:Excitatory Amino Acid Antagonists; D011493:Protein Kinase C; D018091:Receptors, AMPA; D012694:Serine; D013111:Spider Venoms; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; C044936:JSTX spider toxin