Co-administration Of MK-801 Research Articles

The Impact of the Combined Administration of 1MeTIQ and MK-801 on Cell Viability, Oxidative Stress Markers, and Glutamate Release in the Rat Hippocampus

MK-801, as an N-methyl-D-aspartate (NMDA) receptor inhibitor, causes elevation in glutamate release, which may lead to an increase in excitotoxicity, oxidative stress and, consequently, cell death. 1-Methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ) shows antioxidant activity. The aim of the present study was to evaluate the effect of combined treatment with 1MeTIQ and MK-801 on cell viability, antioxidant enzyme activity, and glutamate release in the rat hippocampus. Cytotoxicity was measured using lactate dehydrogenase leakage assay (LDH) and the methyl tetrazolium (MTT) assay; antioxidant enzyme activity (glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD), and catalase (CAT)) were measured by ELISA kits. The release of glutamate in the rat hippocampus was measured using in vivo microdialysis methodology. An in vitro study showed that MK-801 induced cell death in a concentration-dependent manner and that 1MeTIQ partially reduced this adverse effect of MK-801. An ex vivo study indicated that MK-801 produced an increase in antioxidant enzyme activity (GPx, GR, and SOD), whereas coadministration of MK-801 and 1MeTIQ restored the activity of these enzymes to the control level. An in vivo microdialysis study demonstrated that combined treatment with both drugs decreased the release of glutamate in the rat hippocampus. The above results revealed that 1MeTIQ shows limited neuroprotective activity under conditions of glutamate-induced neurotoxicity.

A novel role for involvement of N-methyl-D-aspartate (NMDA) glutamate receptors in sperm acrosome reaction.

Ionotropic glutamate receptors are expressed in mouse and human spermatozoa. However, the possible role of these receptors has not been reported in the sperm acrosome reaction. This study was conducted to demonstrate the function of N-methyl-D-aspartate (NMDA) glutamate receptors in the acrosome reaction of mouse spermatozoa. Epididymal spermatozoa from adult mice were release in a culture medium. The sperm suspension was then divided into six groups: (1) spermatozoa at 0min, (2) spermatozoa at 60min (control), (3) spermatozoa treated with NMDA glutamate receptor agonist (L-glutamate, LG), (4) spermatozoa treated with α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainite glutamate receptor agonist (kainic acid), (5) spermatozoa treated with NMDA glutamate receptor antagonist (MK-801)+LG and (6) spermatozoa treated with ethylene glycol tetraacetic acid (EGTA, as a calcium chelator)+ LG. The sperm samples were examined for the acrosome reaction and intracellular calcium concentration. After 60min, LG but not kainic acid significantly increased both the acrosome reaction and intracellular calcium levels in the spermatozoa compared with the control group. Co-administration of MK-801 or EGTA+LG could significantly reverse the effect of LG in the acrosome reaction and the level of intracellular calcium as compared to the LG group. The possibility that LG induced the acrosome reaction and elevated inter-cellular calcium concentration in mouse spermatozoa and that MK-801 could reverse the effects of LG, may suggest the involvement of NMDA glutamate receptors, at least in the initiation of the acrosome reaction in vitro.

Antidepressant-like effect of ethanol in mice forced swimming test is mediated via inhibition of NMDA/nitric oxide/cGMP signaling pathway.

There is evidence for a dramatic relationship between depression and alcohol consumption. Depressed patients may abuse ethanol because this agent reduces the symptoms of depression. In the current study, we aimed to investigate the NMDA/nitric oxide/cGMP pathway in the antidepressant-like effect of ethanol in an animal model of behavioral despair. Animals were subjected to locomotor activity in an open-field test separately, followed by a forced swimming test. During the forced swimming test (FST), ethanol (2 and 2.5g/kg) significantly decreased the immobility time without altering the locomotor activity of animals. The antidepressant-like effect of ethanol (2.5g/kg) was reversed by co-administration of N-methyl-D-aspartate (NMDA, 75mg/kg), L-arginine (750mg/kg), or sildenafil (5mg/kg). In contrast, co-administration of MK-801 (0.05mg/kg), ketamine (1mg/kg), and ifenprodil (0.5mg/kg) as antagonists of NMDAR, and NG-nitro-L-arginine methyl ester (L-NAME, 10mg/kg), 7-nitroindazole (7-NI, 30mg/kg), and methylene blue (10mg/kg) as inhibitors of nitric oxide synthase (NOS), or 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ) (20mg/kg), a nitric oxide/cyclic-guanosine monophosphate (NO-cGMP) inhibitor, with a subeffective dose of ethanol (1.5g/kg), significantly decreased the immobility time in the FST. Furthermore, injection of ethanol 2.5g/kg alone or 1.5g/kg with a 7-NI subeffective dose, significantly decreased the nitrite levels in the hippocampus and prefrontal cortex. Hence, it is concluded that blockade of NMDA receptors and the nitric oxide/cyclic-guanosine monophosphate (NO-cGMP) pathway might be involved in the antidepressant-like effect of ethanol in mice.

Behavioral effects of combined morphine and MK-801 administration to the locus coeruleus of a rat neuropathic pain model

The persistent activation of N-methyl-d-aspartate acid receptors (NMDARs) seems to be responsible for a series of changes in neurons associated with neuropathic pain, including the failure of opioids that act through mu-opioid receptors (MORs) to provide efficacious pain relief. As the noradrenergic locus coeruleus (LC) forms part of the endogenous analgesic system, we explored how intra-LC administration of morphine, a MORs agonist, alone or in combination with MK-801, a NMDARs antagonist, affects the sensorial and affective dimension of pain in a rat model of neuropathic pain; chronic constriction injury (CCI). Intra-LC microinjection of morphine induced analgesia in CCI rats, as evident in the von Frey and cold plate test 7 and 30 days after surgery, although it was not able to reverse pain-related aversion when evaluated using the place escape/avoidance test. However, the thermal anti-nociception produced by morphine was enhanced when it was administered to the LC of CCI animals in combination with MK-801, without altering its effects on the mechanical thresholds. Furthermore, pain-related aversion was reduced by co-administration of these agents, yet only in the short-term CCI (7 day) rats. Overall the data indicate that administration of morphine to the LC produces analgesia in nerve injured animals and that this effect is potentiated in specific pain modalities by the co-administration of MK-801. While a combination of morphine and MK-801 could reduce pain-related aversion in short-term neuropathic animals, it was ineffective in the long-term, suggesting that its sensorial effects and its influence on the affective component of pain are regulated by different mechanisms.

Involvement of spinal glutamate in nociceptive behavior induced by intrathecal administration of hemokinin-1 in mice

The most recently identified tachykinin, hemokinin-1, was cloned from mouse bone marrow. While several studies indicated that hemokinin-1 is involved in pain and inflammation, the physiological functions of hemokinin-1 are not fully understood. Our previous research demonstrated that the intrathecal (i.t.) administration of hemokinin-1 (0.00625–1.6nmol) dose-dependently induced nociceptive behaviors, consisting of scratching, biting and licking in mice, which are very similar with the nociceptive behaviors induced by the i.t. administration of substance P. Low-dose (0.0125nmol) hemokinin-1-induced nociceptive behavior was inhibited by a specific NK1 receptor antagonist; however, high-dose (0.1nmol) hemokinin-1-induced nociceptive behavior was not affected.In the present study, we found that the nociceptive behaviors induced by hemokinin-1 (0.1 nmol) were inhibited by the i.t. co-administration of MK-801 or D-APV, which are NMDA receptor antagonists. Moreover, we measured glutamate in the extracellular fluid of the mouse spinal cord using microdialysis. The i.t. administration of hemokinin-1 produced a significant increase in glutamate in the spinal cord, which was significantly reduced by co-administration with NMDA receptor antagonists. These results suggest that hemokinin-1-induced nociceptive behaviors may be mediated by the NMDA receptor in the spinal cord.

Inhibitory avoidance memory deficit induced by scopolamine

The possible involvement of N-methyl-D-aspartate (NMDA) receptors of the nucleus accumbens (NAc) in amnesia induced by scopolamine was investigated. An inhibitory (passive) avoidance task was used for memory assessment in male Wistar rats. The results revealed that intra-NAc administration of a nonselective muscarinic acetylcholine antagonist, scopolamine (1 and 2 g/rat) impaired memory consolidation in the animals when tested 24 h later. Post-training intra-NAc administration of NMDA (0.005 and 0.01 g/rat) also impaired memory consolidation, whereas post-training intra-NAc administration of the NMDA receptor antagonist, MK-801 (0.5, 1 and 1.5 g/rat) did not. Intra-NAc co-administration of an ineffective dose of NMDA with ineffective doses of scopolamine (0.25 and 0.5 g/rat) after training had no significant effect on memory consolidation, but intra-NAc injections of effective doses of NMDA (0.005 and 0.01 g/rat) prevented the amnesic effect of an effective dose of scopolamine (2 g/rat). In contrast, intra-NAc co-administration of MK-801 (0.5, 1 and 1.5 g/rat) along with an effective dose of scopolamine (2 g/rat) did not prevent the effect of the latter drug. It can be concluded that NMDA receptors in the NAc are involved in the modulation of memory consolidation that was affected by scopolamine.

Involvement of NMDA receptors in the ventrolateral striatum of rats in apomorphine-induced jaw movements

The role of NMDA receptors in the ventrolateral striatum to modulate dopamine receptor-mediated jaw movements was investigated in freely moving rats, using a magnetic sensor system combined with intracerebral microinjection of drugs. Apomorphine (1 mg/kg i.v.) induced repetitive jaw movements that were reduced, in a dose-dependent manner, by bilateral microinjections of the NMDA receptor agonist NMDA (0.1 and 1 μg/0.2 μl bilaterally) into the ventrolateral striatum. Apomorphine-induced repetitive jaw movements were also reduced, in a dose-dependent manner, by bilateral microinjections of the NMDA receptor antagonists d-APV (0.01 and 0.1 μg) or MK-801 (0.5 and 5 μg). The inhibitory effect of NMDA (1 μg) was reduced by co-administration of MK-801 (0.5 μg). Microinjections of drugs into the ventrolateral striatum in the absence of apomorphine did not affect jaw movements. These results suggest that NMDA receptors in the ventrolateral striatum play an important modulatory role in the expression of dopamine receptor-mediated jaw movements. However, similar effects of NMDA and NMDA antagonists echo previous paradoxical findings and indicate that interactions between dopamine and NMDA receptors are complex and multifaceted. Cellular mechanism(s) may involve differential effects of NMDA agonism and antagonism on dopamine D1-like vs D2-like receptors and, possibly, on related GABAergic processes.

NMDA receptor antagonism disrupts the development of morphine analgesic tolerance in male, but not female C57BL/6J mice

Multiple studies demonstrate that coadministration of N-methyl-D-aspartate (NMDA) receptor antagonists with the opioid agonist morphine attenuates the development of analgesic tolerance. Sex differences in the effects of noncompetitive, but not competitive NMDA receptor antagonists on acute morphine analgesia, have been reported in mice, yet the role of sex in modulation of morphine tolerance by NMDA receptor antagonists has yet to be addressed. Therefore, we tested whether there is a sex difference in the effect of NMDA receptor antagonists on the development of morphine analgesic tolerance in C57BL/6J mice. Acutely, at a dose required to affect morphine tolerance in male mice, the noncompetitive NMDA receptor antagonist dizocilpine (MK-801) prolonged morphine analgesia similarly in both sexes in the hot plate and tail withdrawal assays. In the hot plate assay, coadministration of MK-801 or the competitive antagonist 3-(2-carboxpiperazin-4-yl)propyl-1-phosphanoic acid (CPP) with morphine attenuated the development of tolerance in male mice, while having no effect in females. Like normal and sham females, ovariectomized mice were similarly insensitive to the attenuation of morphine tolerance by MK-801 in the hot plate assay. Surprisingly, in the tail withdrawal assay, MK-801 facilitated the development of morphine-induced hyperalgesia and tolerance in males but not females. The results demonstrate that male mice are more sensitive to modulation of nociception and morphine analgesia after repeated coadministration of NMDA receptor antagonists. Furthermore, the underlying mechanisms are likely to be different from those mediating the sex difference in the modulation of acute morphine analgesia that has previously been reported.

Dynamic changes in N-methyl-D-aspartate receptors after closed head injury in mice: Implications for treatment of neurological and cognitive deficits.

Traumatic brain injury is a leading cause of mortality and morbidity among young people. For the last couple of decades, it was believed that excess stimulation of brain receptors for the excitatory neurotransmitter glutamate was a major cause of delayed neuronal death after head injury, and several major clinical trials in severely head injured patients used blockers of the glutamate N-methyl-D-aspartate (NMDA) receptor. All of these trials failed to show efficacy. Using a mouse model of traumatic brain injury and quantitative autoradiography of the activity-dependent NMDA receptor antagonist MK801, we show that hyperactivation of glutamate NMDA receptors after injury is short-lived (<1 h) and is followed by a profound and long-lasting (> or =7 days) loss of function. Furthermore, stimulation of NMDA receptors by NMDA 24 and 48 h postinjury produced a significant attenuation of neurological deficits (blocked by coadministration of MK801) and restored cognitive performance 14 days postinjury. These results provide the underlying mechanism for the well known but heretofore unexplained short therapeutic window of glutamate antagonists after brain injury and support a pharmacological intervention with a relatively long (> or =24 h) time window easily attainable for treatment of human accidental head injury.

Succinate increases neuronal post-synaptic excitatory potentials in vitro and induces convulsive behavior through N-methyl- d-aspartate-mediated mechanisms

Succinate is a dicarboxylic acid that accumulates due to succinate dehydrogenase inhibition by malonate and methylmalonate exposure. These neurotoxins cause increased excitability and excitotoxic damage, which can be prevented by administering high amounts of succinate. In the present study we investigated whether succinate alters hippocampal field excitatory post-synaptic potentials. Bath application of succinate at intermediate concentrations (0.3–1 mM) increased the slope of field excitatory post-synaptic potentials in hippocampal slices, and at high concentrations (above 1 mM) did not alter or decrease field excitatory post-synaptic potentials slope. Succinate-induced enhancement of field excitatory post-synaptic potentials slope was abolished by the addition of d-2-amino-5-phosphonovaleric acid (50 μM) to the perfusate, supporting the involvement of N-methyl- d-aspartate receptors in the excitatory effect of this organic acid. Accordingly, succinate (0.8–7.5 μmol) i.c.v. administration caused dose-dependent convulsive behavior in mice. The i.c.v. co-administration of MK-801 (7 nmol) fully prevented succinate-induced convulsions, further suggesting the involvement of N-methyl- d-aspartate receptors in the convulsant action of succinate. Our data indicate that accumulation of moderate amounts of succinate may contribute to the excitotoxicity induced by succinate dehydrogenase inhibitors, through the activation of N-methyl- d-aspartate receptors.

D, l- cis-2,3-Pyrrolidine dicarboxylate alters [ 3H]- l-glutamate binding and induces convulsions in mice

This study investigated whether d, l- cis-2,3-Pyrrolidine dicarboxylate ( d, l- cis-2,3-PDC), a new glutamate analogue, alters glutamate binding to cerebral plasma membranes and whether N-methyl- d-aspartate (NMDA) receptors are involved in the convulsant effect of this compound. d, l- cis-2,3-PDC reduced sodium-independent [ 3H]- l-glutamate binding to lysed membrane preparations from adult rat cortex and had no effect on sodium-dependent glutamate binding. Intracerebroventricular administration of d, l- cis-2,3-PDC (7.5–25 nmol/5 μl) induced generalized tonic–clonic convulsions in mice in a dose-dependent manner. The coadministration of MK-801 (7 nmol/2.5 μl), with d, l- cis-2,3-PDC (16.5 nmol/2.5 μl), fully protected the animals against d, l- cis-2,3-PDC-induced convulsions, while the coadministration of DNQX (10 nmol/2.5 μl) increased the latency to convulsions but did not alter the percentage of animals that had convulsions. These results suggest that d, l- cis-2,3-PDC-induced effects are mediated predominantly by NMDA receptors.

N-methyl- d-aspartate-induced excitation and sensitization of normal and inflamed nociceptors

The present study investigates the contribution of peripheral N-methyl- d-aspartate (NMDA) receptors to acute nociception and persistent inflammatory pain in the rat. Immunohistochemical localization of the NMDA receptor one (NMDAR1) subunit demonstrates that 47% of unmyelinated axons in the normal digital nerve are positively labeled. In concert with the overall progression of inflammation following injection of complete Freund’s adjuvant (CFA) in the hind paw, a significant increase in the proportion of NMDAR1-labeled unmyelinated digital axons occurs at 2 and 7, but not 14 days following hind-paw inflammation. In behavioral studies, we confirm an increased mechanical sensitivity in CFA-injected hind paws. Furthermore, activation of NMDA receptors following intraplantar NMDA (1.0 mM) in normal animals results in a mechanical sensitivity similar to that observed in inflamed animals. Conversely, a low concentration of NMDA (0.5 mM) that has little affect on mechanical thresholds in normal animals produces a significant increase in mechanical sensitivity in the inflamed state. CFA-induced mechanical sensitivity involves NMDA-receptor activation demonstrated by the observation that injection of MK-801 alone into the inflamed hind paw returns mechanical sensitivity to normal (pre-inflammation) levels. In single-unit studies, there is a dose-dependent increase in NMDA-induced nociceptor activity in both normal and inflamed skin, but the amount of NMDA required to induce activation is reduced in inflamed skin. In addition, NMDA-induced discharge rates and percentage of NMDA-activated nociceptors are significantly increased in inflamed compared with normal skin, and this activation can be blocked by co-administration of MK-801. Exposure of nociceptors in normal skin to 1 mM NMDA sensitizes the units to reapplication of NMDA and to heat. Nociceptors that demonstrate sensitization to heat in persistent inflammation show an enhanced sensitization when exposed to exogenous NMDA. Thus, peripheral NMDA receptors not only play an important role in modulating the responses of nociceptors in normal skin, but their upregulation and activation on peripheral nociceptors contributes significantly to the mechanical sensitivity and heat sensitization that accompanies persistent inflammation.

Neuroprotective properties of tianeptine: interactions with cytokines

Tianeptine is an antidepressant with proven clinical efficacy and effects on hippocampal plasticity. Hypoxia increased lactate dehydrogenase (LDH) release from cortical neuronal cultures, and tianeptine (1, 10 and 100 μM) inhibited LDH release as efficiently as the N-methyl- d-aspartate (NMDA) antagonist, MK-801. However, tianeptine did not block apoptosis in cultured cortical neurones caused by NMDA, but reduced apoptosis when interleukin-1β (IL-1β) was included with NMDA. In 5-day old mice, intracerebral injection of ibotenate induced reproducible lesions in cortex and white matter. Lesion size was markedly reduced by co-administration of MK-801 (1 mg/kg i.p.) but neither by tianeptine or its enantiomers administered acutely (1, 3 or 10 mg/kg i.p.) nor by tianeptine administered chronically (10 mg/kg i.p. for 5 days). Chronic administration of IL-1β (10 ng/kg i.p. for 5 days) prior to ibotenate injection exacerbated lesion size in cortex and white matter, and this exacerbation was prevented by chronic pre-treatment with tianeptine (10 mg/kg i.p.) or by acute administration of tianeptine (10 mg/kg i.p.) concomitantly with ibotenate. Thus tianeptine has neuroprotective effects against hypoxia in tissue culture and against the deleterious effects of cytokines in cortex and white matter, but not against NMDA receptor-mediated excitotoxicity.

Sedative but not analgesic alpha2 agonist tolerance is blocked by NMDA receptor and nitric oxide synthase inhibitors.

Studies show that the sedative and analgesic effects of alpha2 adrenergic agonists decrease over time, which is a form of synaptic plasticity referred to as tolerance. Because both the N-methyl-D-aspartate (NMDA) receptor complex and nitric oxide synthase are pivotal for some forms of synaptic plasticity, their role in tolerance to the hypnotic and analgesic effects of alpha2 agonists was investigated. After institutional approval, rats were made tolerant to the hypnotic or analgesic action of an alpha2 agonist, dexmedetomidine. The hypnotic response to dexmedetomidine was assessed by the duration of loss of righting reflex, and the analgesic response to dexmedetomidine was assessed by the tail-flick assay. In separate cohorts, either the NMDA receptors or nitric oxide synthase was antagonized by coadministration of MK-801, ketamine, or NO2-arginine, respectively, during induction of tolerance. In a separate series of experiments, after tolerance was induced, the hypnotic and analgesic responses to dexmedetomidine were assessed in the presence of acutely administered MK-801 or NO2-arginine. Induction of tolerance to the hypnotic effect of dexmedetomidine is blocked by coadministration of MK-801, ketamine, and NO2-arginine. However, after tolerance developed, acute administration of MK-801, ketamine, or NO2-arginine did not prevent the expression of tolerance. Coadministration of MK-801 or NO2-arginine neither prevents the development nor reverses the expression of tolerance to the analgesic action of dexmedetomidine. The underlying processes responsible for the development of tolerance to the hypnotic and analgesic actions of systemically administered alpha2 agonists were different, with only the sedative tolerance involving the NMDA receptor and nitric oxide synthase system.

Transient Down-regulation of NMDA Receptor Subunit Gene Expression in the Rat Retina Following NMDA-induced Neurotoxicity is Attenuated in the Presence of the Non-competitive NMDA Receptor Antagonist MK-801

Excitotoxic challenge has been thought to directly target NMDA-receptive neurons to undergo cell death. Recent evidence suggests that NMDA induced cell death is a selective process and that the specificity may be determined by the subunit composition of the NMDA receptor. Using a rat retinal model, we examined the effects of NMDA induced neurotoxicity on the regulation of NMDA receptor subunit gene and protein expression levels to determine if excitotoxic challenge preferentially regulates one or more of the NMDA receptor subunits. Following NMDA insult, the mRNA levels for NR1com, NR2A, NR2B and, to a lesser extent, the NR2C subunit were substantially reduced within 24hr post-treatment (PT), and remained depressed for up to 48hr. Levels for NR2D, although initially suppressed as early as 6hr-PT, were least affected by NMDA insult and showed almost full recovery by 48hr. By 10 days, the levels of gene expression for all five subunits recovered to levels that were indistinguishable from sham treated and untreated retinas. Co-administration of MK-801 with NMDA suppressed the effects of NMDA-induced down-regulation of all five genes.Protein levels for NR1com, NR2A and NR2B were also monitored at select time points following NMDA-insult. By 2 days-PT, protein levels for the three subunits were dramatically reduced. By day 10, the levels of protein expression for NR1comand NR2B remained suppressed despite the rise in gene expression for these two subunits, whereas protein for NR2A showed a substantial rise in expression.Of the five genes assayed, NR2A and NR2B showed the greatest reduction in expression following NMDA treatment, suggesting that one or both of these subunit may signal events leading to neuronal cell death in the retina. Conversely, gene expression of the NR2D subunit was least affected by NMDA exposure. In view of the evidence that the NR2D subunit is expressed by rod bipolar cells in the rat and that these neurons do not die following NMDA insult, it appears that inclusion of this subunit into functional receptors may provide protection against NMDA-induced cell death. Although the significance of the transient down-regulation of four out of the five NMDA receptor subunits is still not fully understood, the recovery of expression of these genes by day 10-PT indicates that not all of the NMDA receptive neurons are susceptible to NMDA-induced cell death. The preferential down-regulation of the NR2A and NR2B receptor subunits may implicate these subunits as key players in mediating the excitotoxic signal in the retina and possibly elsewhere in the brain.

MK-801 potentiates antidystonic effects of clozapine but not of haloperidol in mutant dystonic hamsters

The interaction of nigrostriatal dopamine and corticostriatal glutamate plays a critical role in motor output. Recent studies in mutant dystonic hamsters ( dt sz ), an animal model of idiopathic generalized dystonia, revealed antidystonic effects of both N-methyl- d-aspartate (NMDA) receptor antagonists, such as dizocilpine (MK-801), and of neuroleptics, such as haloperidol and clozapine. Whereas the neuroleptics reduced spontaneous locomotion at antidystonic effective doses, MK-801 caused hyperactivity in mutant hamsters. Therefore, the combination of neuroleptics and MK-801 may exhibit synergistic antidystonic effects, while the side effects may be counteracted. In order to prove this assumption, the neuroleptics haloperidol and clozapine were coadministered with MK-801 in dt sz hamsters in the present study. The antidystonic effects were potentiated by coadministration of MK-801 and clozapine, but not by the combination of MK-801 and haloperidol. The coadministration of MK-801 and clozapine did not cause severe side effects, such as catalepsy in dystonic hamsters. In contrast to the well-documented reverse of haloperidol-induced catalepsy by MK-801 in rats, in mutant hamsters, catalepsy was increased by coadministration of haloperidol and MK-801. This unexpected finding could be due to pathophysiological brain alterations in dt sz hamsters, because in non-dystonic control hamsters, combined treatment with MK-801 and haloperidol did not cause cataleptogenic effects.

Methylation of the NMDA Receptor Agonistl-trans-2,3-Pyrrolidine-Dicarboxylate: Enhanced Excitotoxic Potency and Selectivity

This study investigated the excitotoxic properties of a novel series of NMDA analogues in which a methyl group was introduced to the 5-position of the pyrrolidine ring ofl-trans-2,3-PDC, a previously identified NMDA receptor agonist. While all of these compounds induced NMDA-receptor-mediated injury, methylation increasedin vivoexcitotoxic potency 1000-fold. Injections (1 μl) in rat dorsal hippocampus ofcis- andtrans-5-methyl-l-trans-2,3-PDC (0.1 nmol) induced 50–70% neuronal damage to areas CA1 and CA4, comparable to that induced by 100 nmol ofl-trans-2,3-PDC. Further,cis- andtrans-methylated analogues induced distinct patterns of hippocampal pathology consistent with differential excitotoxic vulnerability of neurons expressing NMDA receptors. Neuronal damage produced by the 5-methyl-l-trans-2,3-PDCs could be blocked by coadministration of MK-801 (3 mg/kg ip), but not NBQX (25 nmol). Biochemical and physiological assays confirmed the action of the analogues as NMDA agonists, but did not provide an explanation for differences in excitotoxic potency between the methylated and nonmethylated 2,3-PDCs. For example, the activity of the compounds as inhibitors of3H-glutamate binding (IC50 values: 0.4, 1.4, and 1.2 μmforcis-5-methyl-,trans-5-methyl-, andl-trans-2,3-PDC, respectively), agonists at NR1A/NR2B receptors (EC50values: 5, 49, and 16 μmforcis-5-methyl-,trans-5-methyl-, andl-trans-2,3-PDC, respectively), andin vitroexcitotoxins in cortical cultures varied only two- to fivefold as a consequence of methylation. Potential roles of NMDA receptor subtypes and transport in these effects are discussed. As potent and selective NMDA excitotoxins,cis- andtrans-5-methyl-l-trans-2,3-PDC will be of value studying excitotoxic mechanisms, NMDA-receptor-mediated pathology, and NMDA receptor heterogeneity.

Contrasting effects of repeated treatment vs. withdrawal of methamphetamine on tyrosine hydroxylase messenger RNA levels in the ventral tegmental area and substantia nigra zona compacta of the rat brain.

We have assessed the effect of repeated treatment with methamphetamine (METH) on the abundance of the messenger ribonucleic acid molecules encoding the enzyme tyrosine hydroxylase (TH) and preprocholecystokinin (PPCCK) in the substantia nigra zona compacta (SNc) and the ventral tegmental area (VTA) by in situ hybridization histochemistry. Rats were injected twice daily with METH (4 mg/kg of body weight) for 6 consecutive days and sacrificed either 5 h or 15 days after the last injection. TH mRNA in the VTA was unaffected by repeated METH treatment but was decreased 25% relative to controls in the SNc. Concurrent administration of METH and MK-801 decreased TH mRNA levels in the SNc to 47% relative to controls. In contrast, TH mRNA levels were found increased in the VTA (42%) but not SNc 15 days post-METH treatment. Coadministration of MK-801 with METH prevented the increase in TH mRNA in the VTA. PPCCK mRNA levels were not significantly affected by METH treatment in VTA or SNc either 5 h or 15 days posttreatment. The results demonstrate that exposure to repeated methamphetamine elicits changes of TH mRNA levels in the VTA that become manifest 2 weeks after withdrawal from this psychostimulant drug.

MK-801 prevents levodopa-induced motor response alterations in parkinsonian rats

The systemic administration of the N-methyl- d-aspartate (NMDA) receptor antagonist, MK801 ((+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine), has previously been found to reverse the motor response alterations that develop during long-term levodopa treatment of parkinsonian rats. To determine whether co-administration of MK801 with levodopa might prevent the initial appearance of these response changes, rats, rendered parkinsonian by a 6-hydroxydopamine lesion of the medial forebrain bundle, received either levodopa alone or levodopa with the NMDA receptor antagonist. After four weeks of treatment with levodopa alone, the duration of the turning response declined by 37% ( P < 0.05) and the number of ineffectual levodopa injections had more than doubled ( P < 0.05). MK801 co-treatment completely blocked the shortening in response duration and prevented the frequency of ineffectual levodopa injection from exceeding baseline levels in animals receiving levodopa alone. The total magnitude of the turning response to levodopa was not affected. These results suggest that NMDA receptor blockade may act prophylactically to prevent the appearance of motor response alterations in levodopa-treated parkinsonian rodents that resemble those occurring in levodopa-treated patients with Parkinson's disease.

Effect of repeated methamphetamine pretreatment on freezing behavior induced by conditioned fear stress

The present study examined the effect of methamphetamine (MA) pretreatment on conditioned fear stress in male Wistar-King rats. Rats received MA or the vehicle according to the repeated escalating dose schedule (1.25, 2.5, 3.75, and 5 mg/kg SC × 2/every other day for a week). After a 5-day drug abstinent period, the rats were exposed to conditioned fear stress (exposure to an environment paired previously with foot shock). Repeated but not single MA pretreatment significantly increased conditioned freezing behavior, suggesting that rats previously exposed to chronic MA are hypersensitive to subsequent stress than control rats. Repeated MA treatment did not decrease basal dopamine or serotonin concentrations in the brain. Furthermore, coadministration of MK-801 (noncompetitive NMDA antagonist), amfonelic acid (dopamine reuptake inhibitor), or fluoxetine (serotonin reuptake inhibitor) with MA did not alter the enhanced freezing behavior. Taken together, it seems that MA-induced hypersensitivity to stress is not due to the neurotoxic effect of MA. Coadministration of nemonapride (D 2/3/4 antagonist) with MA prevented the MA-induced increase in freezing. These results suggest that D 2/3/4 receptors play an important role for MA-induced enhancement of fear or anxiety.