MK-801 Group Research Articles

Effects of MK-801 and morphine on spinal C-Fos expression during the development of neuropathic pain.

The purpose of this study was to investigate the expression of c-fos in the spinal cord during the development of allodynia, induced by peripheral nerve injury. Following tight ligation of the left L5 and L6 spinal nerves of Sprague- Dawley rat, the lumbar spinal cord was postfixed following perfusion. Frontal frozen sections of 40 microm were immunostained according to the peroxidase-antiperoxidase method. The allodynic threshold was checked with 8 calibrated von Frey filaments. MK-801 (0.3 mg/kg), morphine (3 mg/kg) and saline (as a placebo) were administered subcutaneously 30 min before, and 24 and 48 hrs after surgery. The tactile threshold decreased below 3 g since 2 days after surgery in the saline and morphine groups, but delayed a little in the MK-801 group. In the superficial layer the number of Fos-like immunoreactive neurones (Fos-LI) peaked at 2 hours and decreased thereafter, and reached normal levels 24 hrs following operation, for all groups. In the deep layer they were biphasic, - peaking at 2 and 24 hrs - in the saline group, but were suppressed in the morphine and MK-801 groups until 7 days following operation. The above discrepancy between the number of Fos-LI and the allodynic threshold showed that central sensitizations are not critically involved in the development of nerve injury induced tactile allodynia.

Fructose-1,6-bisphosphate and MK-801 by aortic arch flush for cerebral preservation during exsanguination cardiac arrest of 20 min in dogs. An exploratory study

In our exsanguination cardiac arrest (CA) outcome model in dogs we are systematically exploring suspended animation (SA), i.e. preservation of brain and heart immediately after the onset of CA to enable transport and resuscitative surgery during CA, followed by delayed resuscitation. We have shown in dogs that inducing moderate cerebral hypothermia with an aortic arch flush of 500 ml normal saline solution at 4°C, at start of CA 20 min no-flow, leads to normal functional outcome. We hypothesized that, using the same model, but with the saline flush at 24°C inducing minimal cerebral hypothermia (which would be more readily available in the field), adding either fructose-1,6-bisphosphate (FBP, a more efficient energy substrate) or MK-801 (an N-methyl- d-aspartate (NMDA) receptor blocker) would also achieve normal functional outcome. Dogs (range 19–30 kg) were exsanguinated over 5 min to CA of 20 min no-flow, and resuscitated by closed-chest cardiopulmonary bypass (CPB). They received assisted circulation to 2 h, mild systemic hypothermia (34°C) post-CA to 12 h, controlled ventilation to 20 h, and intensive care to 72 h. At CA 2 min, the dogs received an aortic arch flush of 500 ml saline at 24°C by a balloon-tipped catheter, inserted through the femoral artery (control group, n=6). In the FBP group ( n=5), FBP (total 1440 or 4090 mg/kg) was given by flush and with reperfusion. In the MK-801 group ( n=5), MK-801 (2, 4, or 8 mg/kg) was given by flush and with reperfusion. Outcome was assessed in terms of overall performance categories (OPC 1, normal; 2, moderate disability; 3, severe disability; 4, coma; 5, brain death or death), neurologic deficit scores (NDS 0–10%, normal; 100%, brain death), and brain histologic damage scores (HDS, total HDS 0, no damage; >100, extensive damage; 1064, maximal damage). In the control group, one dog achieved OPC 2, one OPC 3, and four OPC 4; in the FBP group, two dogs achieved OPC 3, and three OPC 4; in the MK-801 group, two dogs achieved OPC 3, and three OPC 4 ( P=1.0). Median NDS were 62% (range 8–67) in the control group; 55% (range 34–66) in the FBP group; and 50% (range 26–59) in the MK-801 group ( P=0.2). Median total HDS were 130 (range 56–140) in the control group; 96 (range 64–104) in the FBP group; and 80 (range 34–122) in the MK-801 group ( P=0.2). There was no difference in regional HDS between groups. We conclude that neither FBP nor MK-801 by aortic arch flush at the start of CA, plus an additional i.v. infusion of the same drug during reperfusion, can provide cerebral preservation during CA 20 min no-flow. Other drugs and drug-combinations should be tested with this model in search for a breakthrough effect.

A single high dose of dizocilpine produces long-lasting impairment of the water maze performance in adult rats

The long-lasting effects of a single high dose treatment with the non-competitive N-methyl- d-aspartate (NMDA) antagonist dizocilpine (MK-801; 10 mg/kg, i.p.) on the water maze performance of rats were investigated. MK-801-treated rats learned to find the escape platform at a slower rate than control animals, and showed increased thigmotaxis during acquisition of the task. However, no differences were found between MK-801 and control groups on the probe trial, during repeated acquisition and on the visible platform task. These findings further support the idea that NMDA antagonists can be used in animal research to model persisting symptomatology of cognitive and psychotic disorders.

The Role of the Glutamate Receptor in the Transient Focal Cerebral Ischemia in Rats

Background: To investigate the role of glutamate in the transient focal cerebral ischemia, a reversible middle cerebral artery occlusion madel was induced in 25 male Sprague-Dawley rats. Methods : Triphenyltetrazolium chloride (TTC) stain was used for evaluation of the changes of infarctian ratio in MK-801 (0.3, 1.0, 3.0 mg/kg) or pentylenetetrazole (50 mg/kg) treated groups. Result : The infarction ratio at 48 hours after 2 hour ttansient focal brain ischemia was 39.2 ± 13.2% in control group and 23,8 ± 4.2, 27.0 ±8.9, and 12.8 + 4.4% in MK-801 (0.3, 1.0, 3.0 mg/kg) groups. In the pentylenetetrazole (PTZ) group, the infarction ratio was 32.6 ± 6.7%. Conclusions : The non-specific glutamate receptor antagonist, MK-801, showed a trend toward dose-dependent improvement, but the FIZ group showed no improvement. From these results, it suggested that glutamate might be partly involved in the mechanisms of ischemia-induced neuronal damage. (Korean J Anesthesiol 2000; 38: 1062~1067)

Spinal cord protection: effect of N-methyl-D-aspartate receptor antagonist MK-801 for spinal cord ischemia in a rabbit model.

Excitatory amino acids (glutamate, aspartate) play an important role in the ischemic cascade leading to cell death. The N-methyl-D-aspartate (NMDA) receptor is an excitatory amino acid (EAA) receptor, and NMDA receptor antagonists have been shown to exert a neuroprotective effect in central nervous system ischemia. The purpose of this study was to investigate the effects of noncompetitive NMDA receptor antagonists MK-801 and to observe the changes in EAAs after spinal cord ischemia in a rabbit model. Spinal cord ischemia was induced by clamping the infrarenal abdominal aorta for 24 min. Group 1 (n = 6) received no pharmacologic infusion. Group 2 (n = 5) was administered an intra-aortic hypothermic MK-801 (1 mg/kg) solution and group 3 (n = 6) was administered an intra-aortic normothermic MK-801 (2 mg/kg) solution immediately after clamping of the abdominal aorta. We evaluated the neurological function at 12, 24 and 48 hrs after spinal cord ischemia. A histopathologic study was carried out 72 hrs after spinal cord ischemia, and the results for groups 1 and 3 were compared. The glutamate and aspartate levels in the blood plasma were compared at pre-ischemia and at 12, 24, and 48 hrs among the groups. The perfusion of a normothermic MK-801 (2 mg/kg) solution significantly reduced the neurological dysfunction and the neuronal damage. There was a significant increase in aspartate at 24 and 48 hrs in group 1, but no such increase in glutamate occurred in groups 1 and 3. In conclusion, these data provide the evidence that therapeutic intervention with MK-801 (2 mg/kg) in the early period of spinal cord ischemia is beneficial in reducing neurological dysfunction and neuronal damage.

Effects of MK801 on evoked potentials, spinal cord blood flow and cord edema in acute spinal cord injury in rats

To determine whether MK801, an NMDA receptor antagonist, blocks glutamate excitotoxicity directly or via other mechanisms such as improving blood supply at the injury site in a rat model of spinal cord injury (SCI). In the present study, the effects of pre- and posttreatment with MK801 on axonal function, spinal cord blood flow (SCBF) and cord water content were studied after acute SCI in rats. Somatosensory evoked potentials (SSEPs) and cerebellar evoked potentials (CEPs) were used to quantify electrophysiological function, and the hydrogen clearance technique and wet-dry weight measurements were used to measure SCBF and cord water content, respectively. Twenty rats received a 21 g clip compression injury of the cord at T1, and were then randomly and blindly allocated to either MK801 or saline groups. Each rat received an intravenous infusion of drug or saline four times during the experiment (16 min/infusion) with the first infusion (MK801 3 mg/kg) beginning 8 min pre-injury, and the other infusions (MK801 1. 5 mg/kg) at 1 h intervals after injury. Control experiments on uninjured rats were performed in 10 rats using the same procedure as above except the clip compression injury of the cord was omitted. In the MK801 groups with or without SCI, the amplitude of the evoked potential peaks, especially the SSEPs, was significantly lower than in the saline group. There were no differences in SCBF or cord water content between the MK801 and saline groups. Pre- and posttreatment with MK801 inhibits evoked potentials, but does not improve SCBF or cord edema after acute compression SCI in rats. For the first time it has been shown that MK801 produced a blockade of glutamate excitatory transmission in afferent pathways after SCI. Further work is required to determine whether this inhibition is reversible and related to neuroprotection and functional recovery after SCI.

The Effects of Dexamethasone and MK-801 on Edema Formation in Middle Cerebral Artery Occlusion Model of the Rat

Background: This study was conducted to determine the effects of dexamethasone and MK-801 on the formation of brain edema resulting from a focal ischemic injury by middle cerebral artery occlusion in rats. Methods : Fifteen Sprague-Dawley rats (220 280 g) were freely allowed to drink and eat until just before surgery. Anesthesia was induced with 4% isoflurane in oxygen and then maintained with 2% isoflurane in oxygen. Ischemic injury was induced by an intraluminal suture with a blunted tip inserted into the internal carotid artery and occlusion of the middle cerebral artery. All rats were divided randomly into three groups. In group I (n = 5), normal saline 1 ml was injected intravenously 10 minutes before MCA occlusion. In group II (n = 5), dexamethasone 3 mg/kg was administered 10 minutes before injury. In group III (n = 5), a N-methyl-D-aspartate receptor antagonist, MK-801 1 mg/kg was injected 30 minutes before injury. Rectal temperatures were monitored during the experiment. After 60 minutes of MCA occlusion, the intraluminal sutures were removed and all the rats were returned to their cages. Their brain were quickly removed and the cerebral hemispheres were sepaerated into ischemic cores and penumbra zones after 1 hour of reperfusion. The separated cerebral hemispheres were dried 7 days at 60oC dry oven. The cerebral water content was assessed by the dry-weight method. Results: In the dexamethasone group, there were no significant changes in cerebral edema formation in both ischemic core and the penumbra. In the MK-801 group, there were significant reductions in the brain edema formation in the penumbra (P＜ 0.05), but not in the core. Conclusions : Dexamethasone has no effect on ischemic brain edema; however, MK-801 is effective in the ischemic penumbra zone by reducing of edema formation. (Korean J Anesthesiol 1999; 37: 327∼333)

Isoflurane and Pentobarbital Reduce the Frequency of Transient Ischemic Depolarizations During Focal Ischemia in Rats

Repetitive transient ischemic depolarizations (IDs) during focal cerebral ischemia are thought to contribute to ischemic damage.Isoflurane and pentobarbital reduce injury (versus the nonanesthetized state) after focal cerebral ischemia. The mechanism by which these drugs reduce injury is not known. This protective effect might be mediated by a reduction in the number of IDs. We measured the frequency of IDs during focal cerebral ischemia in animals anesthetized with isoflurane or pentobarbital and compared it with that in N2 O/fentanyl anesthetized animals and in animals in which the N-methyl-D-aspartate receptor antagonist MK801 (dizocilpine) was given. Focal cerebral ischemia was induced by the occlusion of the middle cerebral artery for a period of 2 h. Cortical infarct volumes were determined after 3 h of reperfusion by image analysis of 2,3,5-triphenyl tetrazolium-stained coronal brain sections. The infarct volume was significantly greater in the N2 O/fentanyl group than in the other three groups. Infarct volumes in the isoflurane, pentobarbital, and MK801 groups were similar. The frequency of IDs was significantly greater in the N2 O/fentanyl group than in the other three groups, and was the least in the MK801 group. There was a direct correlation between the number of IDs and the volume of tissue injury. The data indicate that the protective effect of isoflurane and pentobarbital might, in part, be determined by their ability to reduce IDs during focal ischemia. However, the observation that the infarct volume was similar in the MK801, isoflurane, and pentobarbital groups, despite a greater frequency of IDs in the latter two groups, suggests that mechanisms other than a simple reduction in the number of IDs probably also play a role in anesthetic-mediated cerebral protection. Implications: Transient ischemic depolarizations during focal ischemia contribute to brain injury. Both isoflurane and pentobarbital reduced the frequency of these depolarizations. Isoflurane- and pentobarbital-mediated reduction in the frequency of depolarizations might, in part, mediate the previously documented neuroprotective effect of these drugs. (Anesth Analg 1998;86:773-80)

Mitochondrial function and energy metabolism after hypoxia-ischemia in the immature rat brain: involvement of NMDA-receptors.

Treatment after hypoxia-ischemia (HI) in immature rats with the N-methyl-D-aspartate receptor (NMDAR) antagonist dizocilpine maleate (MK-801) reduces areas with high glucose utilization and reduces brain damage. The object was to study the metabolic effects of MK-801 treatment after HI. Seven-day-old rats were randomized to the following groups: non-HI, HI, or HI plus MK-801 (0.5 mg/kg immediately after HI). In the parietal cortex, the mitochondrial respiration was measured in homogenates 1 to 4 hours, and the energy metabolites at 3 and 8 hours after HI. The energy use was calculated from changes in energy metabolites after decapitation at 3 hours after HI. State 3 respiration was reduced by 46%, 32%, and 25% after HI compared with non-HI with pyruvate plus malate, glutamate plus malate, or glutamate plus succinate as substrates, respectively. Uncoupler-stimulated but not state 4 respiration was similarly reduced. The MK-801 augmented pyruvate plus malate-supported state 3 respiration after HI by 42%. The energy utilization was not affected by HI but was reduced by MK-801 treatment in the ipsilateral cortex from 4.6 +/- 2.3 to 2.6 +/- 1.8 micromol high-energy phosphate bond/min/g. The levels of ATP and phosphocreatine did not differ between the HI and HI plus MK-801 groups at 3 hours, but were lower in the HI than in the HI plus MK-801 group at 8 hours after HI. In conclusion, treatment with MK-801 reduced energy utilization and improved mitochondrial function and energy status after HI, suggesting a linkage between NMDAR activation and impaired energy metabolism during reperfusion.

The effect of MK-801 and of brain-derived polypeptides on the development of ischemic lesion induced by photothrombotic occlusion of the distal middle cerebral artery in rats

The effect of neuroprotective drugs on the early and late electrophysiological manifestations of photothrombotic occlusion of distal branches of middle cerebral artery was studied in rats treated with MK-801 and Cerebrolysin (CL). DC potentials were recorded from the irradiated cortex (ischemic core), from the adjacent penumbra zone and from remote intact cortex. Irradiation elicited after a few minutes of spontaneous spreading depression (SD) waves followed during 10–15 min by focal ischemic depolarization (FID) developing in the irradiated cortex and spreading into the perifocal areas. While the core FID amplitude reached about 30 mV and decayed during subsequent 2 h to 10–13 mV, FID in the penumbra zone was broken by periods of partial repolarization and returned during 30–90 min almost to baseline. At the same time, generation of spontaneous SD waves almost stopped. MK-801 (0.5 mg/kg, i.p., 45 min after ischemia) blocked SD waves, but did not shorten penumbra FID, the decay of which was slowed down to the rate found in the ischemic core. CL treatment (2.5 ml/kg, i.p., 1 h after ischemia) did not influence FID in the acute phase of the experiment, but its 10-day administration facilitated post-ischemic recovery indicated by higher amplitude of evoked SD waves penetrating into the former penumbra zone. Morphological examination showed that the volume of total and partial necrosis was increased in the MK-801 group and marginally reduced in the CL group. It is suggested that the absence of the SD-induced hyperperfusion episodes in MK-801-treated rats may accelerate perifocal thrombotization in this model of focal ischemia.

Effects of topical N-methyl-D-aspartate on blood-brain barrier permeability in the cerebral cortex of normotensive and hypertensive rats

This study was performed to examine if blood-brain barrier (BBB) permeability could be increased by N-methyl-D-aspartate (NMDA) in the cerebral cortex, and to compare the degree of alteration of BBB permeability in normotensive and iii chronic hypertensive rats. Twenty- to 22-week-old spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were anesthetized with isoflurane. After craniotomy in 7 animals in each group (SHR and WKY group), an NMOA patch (10 mM) was placed on one cortex (ipsilateral cortex: IC)I and a normal saline patch was placed on the other cortex (control cortex: IC). The other 7 rats in each group were pretreated with MK-801 before placing NMDA and normal saline patches (SHR MK-801 and WKY MK-801 group). The BBB transfer coefficient (ki) was determined using 14C-alpha-aminoisobutyric acid. The mean arterial pressures of the SHR and the SHR MK-801 group were about 65% higher than those of the WKYand the WKY MK-801 groups. In the WKY groupl the Ki of the IC was significantly higher than that of the CC (IC: 1O.O±2.71 CC: 6.2± 2.4 μl g-l min-l). In the WKY MK801 groupl the Ki was similar in both cortices (IC: 8.6± 4.01 CC: 8.2± 3.3). In the SHR groupl the Ki of the IC was significantly higher than that of the CC (IC: 9.5± 3.71 CC: 6.5± 3.4), and the Ki of each cortex was similar to that ofthe corresponding cortex ofthe WKY group. In the SHR MK-B01 groupl the Ki was similar in both cortices (IC: 7.2 ± 1.51 CC: 7.1 ± 2.7), and was also similar to those of the WKY MK-801 group. Our data suggest that NMOA is involved in increasing BBB permeability. In chronic hypertensionl the response of the BBB to NMOA is not altered when compared with normotension. [Neural Res 1997; 19: 539-544]

Effects of excitatory amino acids on cerebral oxygen consumption and blood flow in rat.

This investigation tested the importance of excitatory amino acids' effects on regional cerebral O2 consumption and the concomitant changes in cerebral blood flow (rCBF) in isoflurane anesthetized rats. In the glutamate or N-methyl-D-aspartate (NMDA) groups, 10(-2) M glutamate or NMDA was topically applied to the right cortex and the left cortex was used as a control. One mg/kg dizocilpine maleate (MK-801), a non-competitive NMDA receptor antagonist, was administered (iv) to the MK-801 group and saline was given to the control group. Cortical rCBF was determined using 14C-iodoantipyrine and regional O2 extraction was measured microspectrophotometrically. Cerebral O2 consumption increased 77% after glutamate (contralateral cortex: 9.0 +/- 1.1 ml O2/min/100 g, glutamate treated cortex: 15.9 +/- 3.9), while a 46% increase was observed with the same concentration of NMDA (contralateral cortex: 9.8 +/- 2.0, NMDA treated cortex: 14.3 +/- 5.5). After MK-801, the O2 consumption decreased to 37% of the control value (control cortex: 7.0 +/- 1.3, MK-801 treated cortex: 2.6 +/- 3.9). MK-801 significantly decreased cerebral O2 extraction from 7.1 +/- 1.3 ml O2/100 ml (control cortex) to 5.3 +/- 0.6 (MK-801 treated cortex). However, there was no significant difference in cerebral O2 extraction between treated and contralateral cortex in either the glutamate or NMDA groups. The increase in O2 consumption caused by glutamate or NMDA was coupled with increased rCBF. Glutamate increased rCBF from 95 +/- 5 ml/min/100 g (contralateral cortex) to 165 +/- 31 (treated cortex), while NMDA increased rCBF from 114 +/- 12 (contralateral cortex) to 178 +/- 60 (treated cortex). MK-801 decreased O2 consumption with a lesser decrease of rCBF. The rCBF was 48 +/- 9 in the MK-801 treated cortex and 99 +/- 22 in the control cortex. Some substances produced by the activation of NMDA receptors may be related to the coupling of cerebral metabolism and blood flow, since after blockade of NMDA receptors with MK-801, this relationship is uncoupled. These findings suggest that glutamatergic processes have a major effect on cerebral O2 consumption and that this is at least partly due to NMDA receptors.

The NMDA receptor and cocaine: Evidence that MK-801 can induce behavioral sensitization effects

Antagonism of the NMDA receptor with MK-801 is considered to be an effective pharmacologic manipulation to prevent the development of sensitization effects to drugs such as cocaine. The present study investigated this issue by comparing the behavioral response of separate groups of rats to three treatment cycles of either saline, 0.1 mg/kg MK-801, 10 mg/kg cocaine, or combined MK-801-cocaine ( 0.1 10 mg/kg ). The treatments were spaced 1 week apart and were preceded by two nondrug baseline tests. In the first test cycle, the four groups had equivalent activity levels in the two nondrug tests. In the first drug test only the MK-801-cocaine group exhibited hyperactivity. By the third drug test, the MK-801-cocaine group exhibited an enhanced hyperactivity and the MK-801 group became hyperactive. Thus, behavioral drug sensitization developed but only with groups treated with MK-801. Antagonism of the NMDA receptor under some circumstances can be a highly effective treatment for the induction of behavioral sensitization effects.

Evidence for N-methyl- d-aspartate receptor mediation of cocaine induced corticosterone release and cocaine conditioned stimulant effects

The role of the N-methyl- d-aspartate (NMDA) receptors in cocaine conditioning and sensitization of locomotor activity was studied in four groups of Sprague-Dawley rats. A sub-motoric dose of the NMDA antagonist MK-801 (0.1 mg/kg, i.p.) was employed using a novel dual-compartment Pavlovian drug conditioning paradigm. The animals were placed sequentially in two different test environments in which locomotor activity was monitored. In the first compartment, the animals always received a non-drug test for 20 min. Upon completion of this test, the animals received either saline, cocaine (10 mg/kg i.p.), MK-801 or MK-801 plus cocaine depending on group assignment and were then placed immediately into the second compartment and again tested for 20 min. A total of six non-drug and six drug tests were conducted every other day over a 12-day period. Across all drug/saline treatment and post-treatment tests for conditioning, there were no statistical differences in locomotor activity among the saline and drug treatment groups in the non-drug test environment. In the drug/saline associated environment, however, cocaine had a reliable stimulant effect on locomotion when administered alone or in combination with MK-801. Following a 1-day and again after 21-days of withdrawal, all animals were administered a non-drug test for conditioning in which no injections were administered. On both tests, all groups had equivalent activity levels in the non-drug environment. In the drug/saline environment, only the cocaine group of the three drug treatment groups exhibited conditioned hyperlocomotion. Importantly, MK-801 blocked conditioned hyperlocomotion in the combined cocaine + MK-801 group. MK-801 did not alter serum or brain cocaine concentration or the cocaine effects on dopamine metabolism in limbic brain tissue. The co-administration of MK-801 with cocaine, however, blocked the corticosterone release effect of cocaine. Thus, the NMDA receptor site appears critical for cocaine induced conditioning and for corticosterone release.

The effects of MK-801 injections and dorsal cortex lesions on maze learning in turtles (Chrysemys picta)

The present studies examined the effects of dorsal cortex lesions and injections of MK-801, an NMDA receptor blocker, on learning of a position habit in a maze. In Experiment 1, turtles were trained in a T-maze for food reward. After they had mastered this task, the turtles were randomly assigned to three groups: dorsal cortex lesion, MK-801 injection, or sham-lesion/saline-injection control. The animals were run in an X-maze for 40 days or until they reached criterion. In comparison with controls, the lesion and MK-801 groups were significantly impaired in X-maze learning. Experiment 2 examined the effects of MK-801 injections on measures of general activity. These measures were not affected by injections of MK-801. The results provide evidence for the involvement of the turtle dorsal cortex and the NMDA receptor in the acquisition of a position habit.

Anti-ischaemic efficacy of a nitric oxide synthase inhibitor and a N-methyl-D-aspartate receptor antagonist in models of transient and permanent focal cerebral ischaemia.

1. We have recently developed a new model of transient focal ischaemia in the rat utilising topical application of endothelin-1 to the left middle cerebral artery (MCA). In order to validate this approach the present study assessed the neuroprotective efficacy of the NMDA receptor antagonist dizocilpine (MK-801) in the endothelin-1 model. The anti-ischaemic efficacy of the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) was subsequently evaluated, and contrasted with its efficacy against permanent focal ischaemia, to determine the utility of the endothelin-1 model for identification of novel pharmacoprotective agents. 2. MK-801 (0.12 mg kg-1 bolus, 108 micrograms kg-1 h-1 infusion i.v., either 1 or 2.5 h pre-transient MCA occlusion (MCAO)) induced hypotension that persisted for approximately 1.5 h so that mean arterial blood pressure (MABP) at the time of MCAO was significantly lower in the 1 h group compared with control (MABP: 86 +/- 11, 68 +/- 6 and 84 +/- 4 mmHg (mean +/- s.d.) for saline, 1 h MK-801 and 2.5 h MK-801 groups respectively). The 2.5 h pretreatment schedule resulted in significant reduction (71%) in the volume of hemispheric damage (assessed 4 h post onset of ischaemia) while the 1 h pretreatment schedule did not (volumes of hemispheric damage: 59 +/- 38, 51 +/- 51 and 17 +/- 28 mm3 for saline, 1 h and 2.5 h MK-801 groups). 3. Thus the considerable neuroprotective effect of MK-801 in the endothelin-1 model of transient focal cerebral ischaemia was highly sensitive to drug-induced hypotension. This result is in contrast to previous studies of permanent MCAO where MK-801-induced hypotension did not compromise its neuroprotective action.4. L-NAME (3 mg kg-1, i.v. 30 min pre-MCAO) moderately, but significantly, reduced (16%) the volume of ischaemic damage 4 h post-permanent MCA occlusion, whereas the 29% reduction in volume of damage achieved in the model of transient focal ischaemia did not attain significance due to the greater variability associated with this model. L-NAME did not significantly alter MABP in either model.5. The modest neuroprotection achieved with NO synthase inhibition suggests NO is of relatively minor importance as a mediator of neurotoxicity following permanent focal cerebral ischaemia. In addition the comparable efficacy of L-NAME against transient focal ischaemia suggests the presence of reperfusion does not enhance the contribution of NO to neuronal injury in the acute (4 h) phase following a focal ischaemic insult.

Structural, neurochemical and behavioural consequences of neonatal blockade of NMDA receptor through chronic treatment with CGP 39551 or MK-801

Recent evidence suggests that NMDA receptors may be involved in survival of neurons and establishment of correct connectivity during development. We have treated rat pups from postnatal day 1 to 22 with daily s.c. injections of a competitive (CGP 39551) and a non-competitive (MK-801) antagonist of the NMDA receptor. Body weight of treated rats was decreased by 50–65% at postnatal day 24 and by 25–32% at 70 days of age. Brain weight was decreased by 16–24% at both ages. Among the different brain regions, the cerebellum and striatum appeared more decreased in size than the cortex and hippocampus. Only few minor, and in some cases transient, differences were measured in the cerebellum, the hippocampus and the cortex for a battery of neurochemical markers related to cholinergic, GABAergic and glutamatergic transmission as well as to astrocyte and oligodendrocyte activity. When tested in actometric cages from postnatal days 28 to 60, treated rats exhibited a dramatic increase of spontaneous locomotor activity which was maximal in 28-day-old animals (380% and 250% of control values in CGP 39551 and MK-801 groups, respectively) and was still significant at 60 days of age. Therefore, long-lasting alteration of motor behaviour is obtained by the schedule of chronic treatment adopted for the present experiments. Our results suggest that blockade of NMDA receptors during the critical period of brain maturation may result in permanent alteration of neural circuits.

Pretreatment with MK-801 reduces spontaneous nystagmus following unilateral labyrinthectomy

Unilateral labyrinthectomy results in a syndrome of ocular motor and postural symptoms which abate over time in a process of behavioural recovery known as vestibular compensation. We have previously suggested that an increased Ca 2+ influx in ipsilateral vestibular nucleus (VN) neurons at the time of the unilateral labyrinthectomy may exacerbate the depresssion of VN resting activity caused by the loss of excitatory input from the VIIIth nerve. In order to further test this hypothesis, we administered ( + )-5-methyl-10,11-dihydro-5H-dibcnzo[a,d]cyclohcptcn-5,10-imine hydrogen maleate (MK-801; 1.0 or 2.5 mg/kg i.p.), which blocks Ca 2+ influx via NMDA receptor-mediated ion channels, to guinea pigs 0.5 h before unilateral labyrinthectomy and examined the effects on three symptoms of unilateral labyrinthectomy: spontaneous ocular nystagmus, yaw head tilt and roll head tilt. Pretreatment with MK-801 significantly altered the time course of the vestibular compensation of spontaneous nystagmus and yaw head tilt but had no significant effect on roll head tilt; in particular, 2.5 mg/kg MK-801 depressed spontaneous nystagmus frequency at 10 and 20 h post-labyrinthectomy relative to saline controls ( P < 0.05, post-hoc Scheffé F-test). The reduction in spontaneous nystagmus frequency was not simply a result of extended anesthesia, since other control animals, which received additional injections of the general anesthetic in order to achieve equivalent sleep times to the MK-801 group, did not show reduced spontaneous nystagmus frequency. These results provide preliminary evidence that Ca 2+ influx through NMDA receptor-mediated channels at the time of the unilateral labyrinthectomy may be important to the generation of the ocular motor and postural symptoms which follow, and that recovery from the effects of this Ca 2+ influx may be important to the initiation of vestibular compensation.

Effects of intra-amygdala injections of NMDA receptor antagonists on acquisition and retention of inhibitory avoidance

These experiments examined the effects of intra-amygdala injections of NMDA receptor antagonists on the acquisition and retention of inhibitory avoidance. In Expt. I, rats received bilateral intra-amygdala injections of the NMDA antagonists d,l-AP5 (1–10 μg), d-AP5 (0.03–1 μg), CPP (0.125 or 0.375 μg), or MK-801 (0.2 or 0.5 μg) prior to training in a continuous multiple-trial inhibitory avoidance (CMIA) task. Acquisition of the task was not significantly affected by any of the drug injections. In contrast, all three competitive antagonists, d,l-AP5, d-AP5 and CPP, produced dose-dependent impairment of 48 h retention performance. Although the MK-801 injections did not significantly impair retention performance, the retention scores of the 0.5 μg MK-801 group were bimodally distributed, indicating retention impairment in a subgroup of the animals given that dose. Intra-amygdala injections of 3 or 10 μg d,l-AP5 did not affect footshock sensitivity (Expt. II) or locomotor activity (Expt. III) and their retention-impairing effects were not due to induction of state dependency (Expt. IV). The retention-impairing effects of intra-amygdala injections of NMDA antagonists were not due to diffusion of the drugs dorsally: injections of 1 μg d-AP5 into the striatal area directly above the amygdala impaired acquisition but not retention performance (Expt. V). The retention-impairing effects of 1 μg d-AP5 or 0.5 μg MK-801 were attenuated by giving additional training to the animals shortly after receiving intra-amygdala injections (Expt. VI). The implications of these findings for hypotheses concerning amygdala function in learning and memory are discussed.

MK-801 does not protect against hypoxic-ischemic brain injury in piglets.

The excitatory amino acid inhibitor MK-801 has been shown in many animals species to protect against hypoxic-ischemic brain injury. We sought to determine whether hypoxic-ischemic injury to the newborn pig's brain could be prevented by the use of MK-801. Hypoxic-ischemic injury to the brain was induced in forty 0-3-day-old piglets. They were randomized to receive either 3 mg/kg MK-801 (MK-801 group, n = 20) or vehicle (control group, n = 19) prior to insult. At time 0, the carotid arteries were ligated and the blood pressure was reduced by one third by hemorrhage. At 15 minutes, inspired oxygen was reduced from 50% to 6%. At 30 minutes, inspired oxygen was changed to 100%, carotid ligatures were released, and the withdrawn blood was reinfused. An additional 14 piglets received 3 mg/kg MK-801 but not hypoxic-ischemic injury (drug-only group), and a final group of 11 piglets were subjected to only a sham operation (sham group). Neurological examination scores at 24, 48, and 72 hours showed that MK-801 and drug-only piglets were significantly worse than the controls. Pathological examination of the brains at 72 hours showed significantly greater damage in the brains of the MK-801 and control pigs relative to the sham and drug-only groups. No differences were found between the control and the MK-801 groups. No differences were found between the sham and drug-only groups. MK-801, at a dose of 3 mg/kg, causes neurological dysfunction in piglets lasting at least 72 hours, but neither causes brain damage nor ameliorates the effects of hypoxic-ischemic injury to the brain of the newborn pig.