NMDA causes release of nitric oxide from rat spinal cord in vitro

doi:10.1016/0006-8993(94)91246-7

Brain Research

Volume 637, Issues 1–2, 21 February 1994, Pages 287-291

https://doi.org/10.1016/0006-8993(94)91246-7 Get rights and content

Abstract

Anatomic studies have localized nitric oxide synthase (NOS) activity in the rat spinal cord dorsal horn and intermediolateral cell column. Behavioral and electrophysiologic studies suggest that N-methyl-d-aspartate (NMDA) stimulates nitric oxide synthesis in the dorsal horn. This report describes a novel bioassay to determine directly in vitro whether NMDA causes release of nitric oxide from rat spinal cord. Modified Krebs-Henseleit solution at 26°C was perfused over spinal cord slices from adult male rats, then dropped onto a ring of endothelium-denuded rat aorta. Following preconstriction with phenylephrine, NMDA (10⁻¹⁰ to 10⁻³ M) alone or with other drugs was added to the perfusion solution and vascular tension measured. NMDA-containing solutions applied directly on the preconstricted vessels without exposure to spinal cord tissue had no effect on vessel tone. In contrast, NMDA via the spinal cord perfusion caused concentration-dependent vascular relaxation, which was blocked by MK-801, hemoglobin, methylene blue, and several arginine analogues which inhibit NOS. [¹⁴C]citrulline assay suggested NOS in rat spinal cord was non-endothelial in nature. NMDA perfusion of spinal cord slices in vitro causes vascular relaxation in this bioassay due to actions on NMDA receptors and which is consistent with release of nitric oxide. These results support previous anatomical, behavioral, and electrophysiologic studies in rat spinal cord and describe a novel, sensitive, and simple bioassay for nitric oxide release from neural tissue in vitro.

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Cited by (52)

Histaminergic Receptors Modulate Spinal Cord Injury-Induced Neuronal Nitric Oxide Synthase Upregulation and Cord Pathology: New Roles of Nanowired Drug Delivery for Neuroprotection
2017, International Review of Neurobiology
Citation Excerpt :
Thus, NO-induced increased histamine outflow after blockade of M1 receptors is caused by activation of glutamate (Broussard, Bao, Li, & Altschuler, 1995; Prast & Philippu, 2001). Thus, activation of excitatory amino acid receptors, e.g., N-methyl-d-aspartate (NMDA) and kainate, enhance histamine release in the hypothalamus (Broussard et al., 1995; Li, Tong, Eisenach, & Figueroa, 1994). The histamine release by activation of NMDA is abolished by tetrodotoxin is in line with the idea that glutamate is responsible for the enhanced histamine release by NO donors after blockade of M1 receptors (see Philippu, 2016; Prast & Philippu, 2001).
The possibility that histamine influences the spinal cord pathophysiology following trauma through specific receptor-mediated upregulation of neuronal nitric oxide synthase (nNOS) was examined in a rat model. A focal spinal cord injury (SCI) was inflicted by a longitudinal incision into the right dorsal horn of the T10–11 segments. The animals were allowed to survive 5 h. The SCI significantly induced breakdown of the blood–spinal cord barrier to protein tracers, reduced the spinal cord blood flow at 5 h, and increased the edema formation and massive upregulation of nNOS expression. Pretreatment with histamine H1 receptor antagonist mepyramine (1 mg, 5 mg, and 10 mg/kg, i.p., 30 min before injury) failed to attenuate nNOS expression and spinal cord pathology following SCI. On the other hand, blockade of histamine H2 receptors with cimetidine or ranitidine (1 mg, 5 mg, or 10 mg/kg) significantly reduced these early pathophysiological events and attenuated nNOS expression in a dose-dependent manner. Interestingly, TiO₂-naowire delivery of cimetidine or ranitidine (5 mg doses) exerted superior neuroprotective effects on SCI-induced nNOS expression and cord pathology. It appears that effects of ranitidine were far superior than cimetidine at identical doses in SCI. On the other hand, pretreatment with histamine H3 receptor agonist α-methylhistamine (1 mg, 2 mg, or 5 mg/kg, i.p.) that inhibits histamine synthesis and release in the central nervous system thwarted the spinal cord pathophysiology and nNOS expression when used in lower doses. Interestingly, histamine H3 receptor antagonist thioperamide (1 mg, 2 mg, or 5 mg/kg, i.p.) exacerbated nNOS expression and cord pathology after SCI. These novel observations suggest that blockade of histamine H2 receptors or stimulation of histamine H3 receptors attenuates nNOS expression and induces neuroprotection in SCI. Taken together, our results are the first to demonstrate that histamine-induced pathophysiology of SCI is mediated via nNOS expression involving specific histamine receptors.
Intrathecal morphine-3-glucuronide-induced nociceptive behavior via Delta-2 opioid receptors in the spinal cord
2016, Pharmacology Biochemistry and Behavior
Citation Excerpt :
On the basis of our previous results, glutamate released from presynaptic sites in response to i.t. M3G activated NMDA receptors, which triggered a feed-forward mechanism of stimulation of nNOS activity via a mechanism that is largely dependent on calcium. In addition, activation of NMDA receptors can stimulate nNOS activity via a calcium calmodulin-dependent mechanism (Li et al., 1994, Wu et al., 1994). Extracellular calcium also appears essential for noxious stimulation-induced ERK activation (Lever et al., 2003).
Intrathecal (i.t.) injection of morphine-3-glucuronide (M3G), a major metabolite of morphine without analgesic actions, produces severe hindlimb scratching followed by biting and licking in mice. The M3G-induced behavioral response was inhibited dose-dependently by pretreatment with an antisera against dynorphin. However, the selective κ-opioid receptor antagonist, nor-BNI did not prevent the M3G-induced behavioral response. Dynorphin is rapidly degraded by a dynorphin-converting enzyme (cystein protease), to leucine-enkephalin (Leu-ENK). The M3G-induced behavioral response was inhibited dose-dependently by pretreatment with the antisera against Leu-ENK. We also showed that M3G co-administered with Leu-ENK-converting enzyme inhibitors, phosphoramidon and bestatin produced much stronger behavioral responses than M3G alone. Furthermore, the M3G-induced behavioral responses were inhibited dose-dependently by i.t. co-administration of the non-selective δ-opioid receptor antagonist, naltrindole or the selective δ₂-opioid receptor antagonist, naltriben, whereas the selective δ₁-opioid receptor antagonist, BNTX had no effect. An i.t. injection of M3G also produced a definite activation of ERK in the lumbar dorsal spinal cord. Western blotting analysis revealed that antisera against dynorphin, antisera against Leu-ENK, naltrindole or naltriben resulted in a significant blockade of ERK activation induced by M3G in the spinal cord. Taken together, these results suggest that M3G-induced nociceptive responses and ERK activation may be triggered via δ₂-opioid receptors activated by Leu-ENK, which is formed from dynorphin in the spinal cord.
Spinal ERK2 activation through δ<inf>2</inf>-opioid receptors contributes to nociceptive behavior induced by intrathecal injection of leucine-enkephalin
2014, Peptides
Citation Excerpt :
Therefore, these results suggest that activation of spinal nNOS-NO pathway contribute to nociceptive behavior induced by i.t. Leu-ENK in combination with peptidase inhibitor. Activation of NMDA receptors has been shown in several studies to stimulate nNOS activity, through a calcium–calmodulin dependent mechanism [26,47]. It appears that glutamate released from presynaptic sites in response to i.t. Leu-ENK in combination with peptidase inhibitors activates the NMDA receptor, which trigger a feed forward mechanism of stimulation of nNOS activity via a mechanism largely dependent on calcium.
Intrathecal (i.t.) injection of leucine-enkephalin (Leu-ENK), co-administered with peptidase inhibitors, phosphoramidon (an endopeptidase 24.11 inhibitor), and bestatin (a general aminopeptidase inhibitor), produced behaviors consisting of the biting and/or licking of the hindpaw and the tail along with hindlimb scratching directed toward the flank, which peaked at 10–15 min after an injection. This characteristic behavior was not observed in mice treated with i.t. Leu-ENK alone. We also investigated the effect of the extracellular signal-regulated kinase (ERK) in spinal processing of nociception induced by i.t. co-administration of Leu-ENK with phospharamidon and bestatin. Western blot analysis of phospho-ERK (pERK) showed a significant increase of pERK2 in the lumbar spinal cord in response to i.t. Leu-ENK co-injected with peptidase inhibitors. The MAP kinase-ERK inhibitor, U0126 dose-dependently attenuated the nociceptive behavior and spinal ERK activation to i.t. Leu-ENK co-injected with peptidase inhibitors. Furthermore, the nociceptive behavior and spinal ERK activation evoked by i.t. Leu-ENK in combination with peptidase inhibitors were inhibited by co-administration of the non-selective δ-opioid receptor antagonist, naltrindole, the selective δ₂-opioid receptor antagonist, naltriben, the non-competitive N-methyl-d-aspartate (NMDA) antagonist, MK-801 or the non-selective nitric oxide synthase inhibitor, l-NAME, the selective nNOS inhibitor, N^ω-propyl-l-arginine or the selective iNOS inhibitor, W1400, but not by the selective δ₁-receptor antagonist, BNTX (7-benzylidenenaltrexone). These results suggest that spontaneous nociceptive behaviors produced by i.t. co-administration of Leu-ENK with peptidase inhibitors may be induced by an activation of the glutamate-NO-ERK pathway through the δ₂-opioid receptor in the dorsal spinal cord.
The cannabinoid 1 receptor antagonist AM251 produces nocifensive behavior via activation of ERK signaling pathway
2010, Neuropharmacology
Citation Excerpt :
The NK1 and NMDA receptor antagonists were significantly reduced inflammation-induced iNOS activity (Anna and Luigi, 2000; Munhoz et al., 2008). The activation of NMDA receptors has been shown to stimulate nNOS activity, through a Ca2+/calmodulin dependent mechanism (Li et al., 1994; Wu et al., 1994). NK1 receptor activates phospholipase C to produce inositol-1,4,5-trisphosphate and increase in intracellular Ca2+ channels (Mochizuki-Oda et al., 1994).
Intrathecal (i.t.) injection of AM251, a cannabinoid 1 (CB₁) receptor antagonist, into the spinal lumbar space of mice elicited a behavioral response consisting of biting and licking with a few scratchings. In this study, we investigated to determine whether i.t. AM251 could influence the activity of extracellular signal-regulated kinase-1 and -2 (ERK1/2), a mitogen-activated protein kinase (MAPK) in neuronal nitric oxide synthase (nNOS) and inducible NOS (iNOS) activation. The CB₁ receptor agonist ACEA, neurokinin 1 (NK₁) receptor antagonists and NMDA receptor antagonists, inhibited i.t. AM251-induced behavioral response in a dose-dependent manner. The CB₂ receptor agonist, JWH-133 gave no effect on response elicited by i.t. AM251. Both non-selective NOS inhibitors, l-NAME and 7-NI, and N^ω-propyl-l-arginine, a selective inhibitor of nNOS resulted in a dose-dependent inhibition of i.t. AM251-induced behavioral response. The selective iNOS inhibitor, 1400W, in relatively large doses, inhibited in a non dose-dependent manner. The i.t. injection of AM251 produced a definite activation of ERK1/2 in the lumbar dorsal spinal cord. Behavioral experiments showed that U0126, a MAPK/ERK kinase (MEK) inhibitor, dose-dependently attenuated the behavioral response to i.t. AM251. Spinal activation of ERK1/2 following i.t. AM251 was reduced clearly by N^ω-propyl-l-arginine and U0126, while 1400W gave a significant effect on only ERK1 activation. These findings suggest that the nNOS–ERK pathway in spinal cord neurones plays an important role in AM251-induced nocifensive behavior and its inhibition may provide significant anti-nociception.
Extracellular signal-regulated kinase (ERK) and nitric oxide synthase mediate intrathecal morphine-induced nociceptive behavior
2007, Neuropharmacology
Intrathecal (i.t.) administration of morphine at a high dose of 60 nmol into the spinal lumbar space in mice produces a severe hindlimb scratching followed by biting and licking. Nitric oxide (NO) is thought to play an important role in signal transduction pathways that enhance nociceptive transmission in the spinal cord. The present study was designed to determine whether high-dose i.t. morphine could influence the activation of the extracellular signal-regulated kinase (ERK), a mitogen-activated protein (MAP) kinase in neuronal nitric oxide synthase (nNOS) and inducible NOS (iNOS) activation. Both 7-NI and TRIM, selective inhibitors of nNOS, resulted in a dose-dependent inhibition of high-dose i.t. morphine-induced behavior. The selective iNOS inhibitor W1400 in relatively large doses inhibited in a non dose-dependent manner. The i.t. injection of morphine evoked a definite activation of ERK in the lumbar dorsal spinal cord. Behavioral experiments showed that U0126 (0.5–2.5 nmol), a MAP kinase-ERK inhibitor, dose-dependently attenuated the behavioral response to i.t. morphine. In mice treated with high-dose morphine, 7-NI was very effective in blocking ERK activation, whereas W1400 had no effect. Taken together, these results suggest that the behavioral response to high-dose i.t. morphine may be triggered by the nNOS-ERK pathway in the dorsal spinal cord.
Spinal modulation of the muscle pressor reflex by nitric oxide and acetylcholine
2000, Brain Research Bulletin
Static contraction of skeletal muscle activates the sympathetic nervous system, which in turn increases cardiovascular function. These changes are mediated, in part, by a reflex arising from the contracting muscle. This reflex is termed the exercise pressor reflex or, more simply, the muscle pressor reflex (MPR). Over the past few years, studies have been performed investigating the sensory processing that occurs in the dorsal horn of the spinal cord as it pertains to the MPR. Several putative neurotransmitters and receptors have been implicated in mediating the MPR at the level of the dorsal horn. In addition, several receptor systems have been shown to modulate the MPR at the dorsal horn. We have recently performed studies investigating the potential modulatory role of dorsal horn nitric oxide (NO) and acetylcholine (ACH) on the MPR. Along these lines, our experiments suggest that NO enhances the excitability of dorsal horn cells receiving input from muscle afferent neurons, while ACH decreases the MPR when its concentration in the dorsal horn is elevated. The purpose of this manuscript is to review recently published findings from our laboratory and apply this information in an effort to better understand the integration of sensory input that occurs in the dorsal horn as it pertains to cardiovascular regulation. This review is also designed to stimulate questions as to how these two neurochemicals exert their actions and whether or not they represent or can represent important physiological mechanisms involved in regulating the dorsal horn integration of the MPR.

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NMDA causes release of nitric oxide from rat spinal cord in vitro

Abstract

Neurosci. Lett.

Anal. Biochem.

Neuron

Pain

Pain

Neurosci. Lett.

Neuroscience

Neuropharmacology

Brain Res.

Brain Res.

Eur. J. Pharmacol.

Pain