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NEUROHUMORAL CONTROL OF CARDIOVASCULAR FUNCTION

Calcium/calmodulin-dependent kinase II mediates NO-elicited PKG activation to participate in spinal reflex potentiation in anesthetized rats

Departments of ¹Obstetrics and Gynecology and ²Ophthalmology, Chung Shan University Hospital; Departments of ³Physiology and ⁵Anatomy, College of Medicine, Chung Shan Medical University, Taichung, Taiwan; ⁴School of Physical Therapy, College of Medicine, China Medical University, Taichung, Taiwan; Departments of ⁶Biotechnology and ⁷Medical Engineering, Ming Chuan University, Taoyuan, Taiwan; ⁸Department of Veterinary Medicine, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan; and ⁹Medical Department, Saint Paul's Hospital, Taoyuan, Taiwan

Submitted 20 August 2007 ; accepted in final form 20 November 2007

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, N^G-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 analog, 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 hyperalgesia and allodynia.

spinal reflex potentiation; soluble guanylate cyclase; cyclic monophosphate sodium salt monohydrate; spinal cord; windup

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