Nitric oxide (NO) release by glutamate and NMDA in the dorsal horn of the spinal cord: an in vivo electrochemical approach in the rat

Abstract Glutamate acts as a neurotransmitter of primary afferent messages in the spinal cord. Through glutamatergic mechanisms nitric oxide (NO) is also a potential intermediary in the transmission of sensory messages, particularly nociceptive, at the spinal level. The aim of the present study was, by using electrochemical monitoring of NO, to determine if the activation of glutamatergic transmission, particularly through NMDA receptors, could increase NO production within the dorsal horn of the lumbar spinal cord in the rat. 30 μm diameter treated carbon fiber electrodes coated with nickel–porphyrine and Nafion®, and associated with differential normal pulse voltammetry, have been used in vivo to monitor NO within the dorsal horn of the lumbar spinal cord of decerebrated–spinalized rats. A NO-dependent peak of oxidation current (650 mV vs. Ag–AgCl), remaining stable for up to 3 h (±5%) could be detected under basal conditions, which indicates that significant amounts of NO are produced continuously. The non-competitive N -methyl- d -aspartate (NMDA) channel blockers, Ketamine (100 mg kg −1 i.p.) and MK-801 (10 mg kg −1 i.p.), decreased the voltammograms to 70±5% and 69±2% of controls at 120 min, respectively. Glutamate (10 mM), when directly superfused upon the spinal cord (20 min at 50 μl min −1 ) induced a rapid and significant increase of the 650 mV peak, with a maximum at around 90 min (148±6% of control) followed by a slow decay (138±4% of control at 150 min). This increase could be totally reversed or blocked by i.p. injection of 100 mg kg −1 of Ketamine. NMDA (30 mg kg −1 i.p.) induced a long-lasting increase in the peaks (149±11% at 90 min and 162±20% at 120 min), which was also fully reversed by Ketamine or MK-801. These results provide in vivo direct evidence of a glutamate- and/or NMDA-induced release of NO at the spinal level, and is discussed in relation to the glutamatergic transmission of primary afferent messages.