Involvement of NO-guanylyl cyclase pathway for the depression of spinal monosynaptic reflex by Mesobuthus tamulus venom in neonatal rat in vitro.

Aims: The present study was undertaken to evaluate the role of nitric oxide (NO) in Mesobuthus tamulus (MBT) venom-induced depression of spinal reflexes.

Main Methods: Experiments were performed on isolated hemisected spinal cords from 4 to 6day old rats. Stimulation of a dorsal root with supramaximal strength evoked monosynaptic (MSR) and polysynaptic reflex (PSR) potentials in the corresponding segmental ventral root.

Key Findings: Superfusion of MBT venom (0.3μg/ml) depressed the spinal reflexes in a time-dependent manner and the maximum depression was seen at 10min (MSR by 63%; PSR by 79%). The time to produce 50% depression (T-50) of MSR and PSR was 7.7±1.3 and 5.7±0.5min, respectively. Pretreatment with bicuculline (1μM; GABA(A) receptor antagonist) or strychnine (1μM; glycine(A) receptor antagonist) did not block the venom-induced depression of spinal reflexes. However, Nω-nitro-L-arginine methyl ester (L-NAME, 100 or 300μM; NO synthase inhibitor) or hemoglobin (Hb, 100μM; NO scavenger) antagonized the venom-induced depression of MSR. Further, soluble guanylyl cylase inhibitors (1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one, ODQ; 1μM or methylene blue, 100μM) also antagonized the venom-induced depression of MSR but not PSR. Nitrite concentration (indicator of NO activity) of the cords exposed to venom (0.3μg/ml) was not different from the control group.

Significance: The results indicate that venom-induced depression of MSR is mediated via NO-guanylyl cyclase pathway without involving GABAergic or glycinergic system.