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1 School of Physical Therapy, China Medical University, 91 Hsueh-Shih Rd., Taichung, 40202, Taiwan - Republic of China
2 Department of Veterinary Medicine, National Chung Hsing University, 8F.-2, No.86, Jhongsiao Rd., South District, Taichung City 402, Taiwan (R.O.C.), Taichung, Taiwan - Republic of China; , Taiwan - Republic of China
3 Department of Physiology, College of Medicine, Chung-Shan Medical University, Tai-Chung, United States
* To whom correspondence should be addressed. E-mail: tblin{at}csmu.edu.tw.
Calcium/Calmodulin protein kinase (CaMK)-dependent nitric oxide (NO) and the downstream intracellular messenger cGMP, which is activated by soluble guanylate cyclase (sGC), are believed to induce long-term changes in efficacy of synapses through the activation of protein kinase G (PKG). The aim of this study was to examine the involvement of the CaMKII-dependent NO/sGC/PKG pathway in a novel form of repetitive stimulation-induced spinal reflex potentiation (SRP). A single pulse test stimulation (TS, 1/30 Hz) on the afferent nerve evoked a single action potential, while repetitive stimulation (RS, 1 Hz) induced a long lasting SRP that was abolished by a selective Ca2+/CaMKII inhibitor, autocamtide 2-related inhibitory peptide (AIP). Such an inhibitory effect was reversed by a relative excess of nitric oxide synthase (NOS) substrate, L-arginine. In addition, the RS-induced SRP was abolished by pretreatment with the NOS inhibitor, NG-nitro-L-arginine-methyl ester (L-NAME). The sGC activator, protoporphyrin IX (PPIX), reversed the blocking effect caused by L-NAME. On the other hand, a sGC blocker, 1H- [1, 2, 4] oxadiazolo [4, 3 - 
] quinoxalin -1-one (ODQ), abolished the RS-induced SRP. Intrathecal applications of the membrane-permeable cGMP analogue, 8-Bromoguanosine 3, 5-cyclic monophosphate sodium salt monohydrate (8-Br-cGMP), reversed the blocking effect on the RS-induced SRP elicited by the ODQ. Our findings suggest that a CaMKII-dependent NO/sGC/PKG pathway is involved in the RS-induced SRP, which has pathological relevance to hypergesia and allodynia.
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