Potentiation of spinal NMDA-mediated nociception by cocaine- and amphetamine-regulated transcript peptide via PKA and PKC signaling pathways in rats.
Our previous study showed that cocaine- and amphetamine-regulated transcript peptide fragment 55-102 (CARTp) specifically potentiated spinal N-methyl-D-aspartate (NMDA)-mediated nociceptive transmission in vivo and in vitro. The cellular mechanisms underlying CARTp potentiation of NMDA receptor function remains unclear. The present study was carried out to test the hypothesis that CARTp changes the phosphorylated state of NMDA receptors by activating intracellular signals and subsequently increasing the function of NMDA receptors. We found that the potentiating effect of CARTp on spinal NMDA-induced hyperalgesia in rats was reduced by intrathecal pretreatment with KT5720 (a selective PKA inhibitor) or GF109206X (a selective PKC inhibitor), but was increased by pretreatment with calyculin A (a protein phosphatase inhibitor). In the in vitro electrophysiological study, CARTp potentiation of NMDA-induced depolarizations was blocked by superfusion of PKA or PKC inhibitor applied 10 min before the application of CARTp. The levels of phosphoserine 897 on the NR1 subunit (pNR1-ser897) and phosphoserine 896 on the NR1 subunit (pNR1-ser896) in the dorsal horn of spinal lumbar segments significantly increased following CARTp superfusion in vitro or intrathecal injection in vivo. The increases in pNR1-ser897 and pNR1-ser896 in the in vivo and in vitro studies were inhibited by pretreatment with KT5720 and GF109206X, respectively. The results provide the first evidence that CARTp increases the phosphorylation of NMDA receptor NR1 subunit via activation of PKA and PKC signals, which may play a crucial role in CARTp regulation of spinal NMDA receptor-mediated nociceptive responses.
Chiu
Hong-Yi
HY
Institute of Medical Sciences, Tzu Chi University, Hualien 970, Taiwan.
Lin
Hsun-Hsun
HH
Lai
Chih-Chia
CC
D000818:Animals; D015153:Blotting, Western; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D016202:N-Methylaspartate / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*; D010146:Pain / Q000378:metabolism*; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013131:Spine / Q000201:enzymology / Q000378:metabolism*; D000098882:Cocaine- and Amphetamine-Regulated Transcript Protein
D009419:Nerve Tissue Proteins; D000098882:Cocaine- and Amphetamine-Regulated Transcript Protein; D010768:Phosphoserine; D016202:N-Methylaspartate; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C