dizocilpine-maleate and Brain-Concussion

Endogenous cannabinoid receptor ligands (endocannabinoids) may rescue neurons from glutamate excitotoxicity. As these substances also accumulate in cultured immature neurons following neuronal damage, elevated endocannabinoid concentrations may be interpreted as a putative neuroprotective response. However, it is not known how glutamatergic insults affect in vivo endocannabinoid homeostasis, i.e. N-arachidonoylethanolamine (anandamide) and 2-arachidonoylglycerol (2-AG), as well as other constituents of their lipid families, N-acylethanolamines (NAEs) and 2-monoacylglycerols (2-MAGs), respectively. Here we employed three in vivo neonatal rat models characterized by widespread neurodegeneration as a consequence of altered glutamatergic neurotransmission and assessed changes in endocannabinoid homeostasis. A 46-fold increase of cortical NAE concentrations (anandamide, 13-fold) was noted 24 h after intracerebral NMDA injection, while less severe insults triggered by mild concussive head trauma or NMDA receptor blockade produced a less pronounced NAE accumulation. By contrast, levels of 2-AG and other 2-MAGs were virtually unaffected by the insults employed, rendering it likely that key enzymes in biosynthetic pathways of the two different endocannabinoid structures are not equally associated to intracellular events that cause neuronal damage in vivo. Analysis of cannabinoid CB(1) receptor mRNA expression and binding capacity revealed that cortical subfields exhibited an up-regulation of these parameters following mild concussive head trauma and exposure to NMDA receptor blockade. This may suggest that mild to moderate brain injury may trigger elevated endocannabinoid activity via concomitant increase of anandamide levels, but not 2-AG, and CB(1) receptor density.

Topics: Animals; Arachidonic Acids; Brain Concussion; Cannabinoid Receptor Modulators; Cerebral Cortex; Corpus Striatum; Craniocerebral Trauma; Dizocilpine Maleate; Endocannabinoids; Ethanolamines; Excitatory Amino Acid Agonists; Excitatory Amino Acid Antagonists; Glycerides; Male; N-Methylaspartate; Nerve Degeneration; Polyunsaturated Alkamides; Rats; Rats, Sprague-Dawley; Receptors, Cannabinoid; Receptors, Drug; RNA, Messenger

The authors studied the effect of a mild cortical contusion to the rat brain on behavioural and morphological outcome and the influence of NMDA-receptor blockade (MK-801, 0.5 mg/kg i.v. 30 min prior to trauma). Spontaneous motor activity was assessed 16-18 days post trauma. Saline treated traumatised rats showed a significant (p < 0.01) hyperactive behaviour compared to animals without injury. MK-801 treated rats performed significantly better than the saline treated animals (p < 0.05). For histopathological evaluation hippocampal hilar neurons were counted, cortical thickness under the impact was measured and microtubule-associated protein 2 (MAP2) immunoreactivity in the dentate hilus was quantified 1, 3 and 21 days post trauma. In traumatised rats scattered loss of nerve cells, oedema and minute haemorrhages were present at the site of the impact one and three days after injury. At day 21 there was a significant reduction of cortical thickness at the site of impact. One day after trauma there was a bilateral, significant loss of neurons and MAP2 immunostaining in the dentate hilus of the hippocampus. MK-801 pretreated rats showed similar morphological changes. The disturbed spontaneous motor behaviour may be caused by hippocampal damage and a reduction of somatosensory cortical neurons. NMDA-receptor blockade improved the outcome assessed by the functional tests but failed to influence the morphological changes, suggesting that this behavioural test is a more sensitive indicator of outcome after mild traumatic brain injury (TBI).

Topics: Animals; Behavior, Animal; Brain Concussion; Cerebral Cortex; Dizocilpine Maleate; Male; Neurologic Examination; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate

Free radicals, lipid peroxidation and excitatory amino acids have been implicated in the secondary mechanisms of traumatic brain injury. We used the cold injury model in rats to assess the endogenous activity of the protective enzyme superoxide dismutase (SOD) and the lipid peroxidation level in the contused tissue at an early phase of injury. Furthermore, we treated the rats with two different N-methyl-D-aspartate receptor antagonists, namely MK-801 and CPP, and evaluated their effect on lipid peroxidation in the contused tissue. Rats were divided into four groups: sham, control, treatment 1 and treatment 2 groups (n= 16 for each group). Thirty and 60 min after craniectomy or injury, tissue samples were removed. SOD activity didn't change in this period. However, lipid peroxidation in terms of malondialdehyde (MDA) amount showed a significant increase at 60 min. Fifteen minutes after injury, MK-801 (1 mg/kg), CPP (10 mg/kg) or saline (1 ml) were applied intraperitoneally in treatment 1, treatment 2 and the control groups. Treatment with MK-801 attenuated MDA levels, whereas treatment with CPP did not. The protective effect of MK-801 achieved statistical significance. These results demonstrate that SOD activity does not change in the early period of cold injury. Moreover, these results show that lipid peroxidation increases after 60 min of cold injury, and treatment with MK-801 15 min after injury can prevent this elevation.

Topics: Animals; Brain Concussion; Cold Temperature; Disease Models, Animal; Dizocilpine Maleate; Excitatory Amino Acid Antagonists; Free Radicals; Hypothermia; Lipid Peroxidation; Male; Piperazines; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate; Superoxide Dismutase

The authors have examined the effect of experimental traumatic brain injury on the amnesia produced by the N-methyl-D-aspartate (NMDA) antagonist MK-801. Rats were either subjected to a moderate level of fluid-percussion injury or prepared for injury but not injured ("sham injury"). Nine days following injury or sham injury, the rats were injected either with saline (sham/saline group, nine rats; injured/saline group, nine rats) or with 0.1 mg/kg of MK-801 (sham/MK-801 group, nine rats; injured/MK-801 group, eight rats) 30 minutes before being trained on a passive-avoidance task. Twenty-four hours later, the rats were tested for retention of the passive-avoidance task. Results revealed that the low dose of MK-801 did not significantly affect retention of the passive-avoidance task in the sham-injured group. In injured animals, administration of MK-801 produced a profound amnesia in contrast to the sham-injured animals treated with MK-801 and the injured animals treated with saline. To further investigate this enhanced sensitivity to the amnesic effects of MK-801 exhibited by the injured animals, nine injured and eight sham-injured rats were injected with 0.3 mg/kg of MK-801 15 minutes before injury. Results indicated that the animals treated with MK-801 before injury did not significantly differ from the sham-injured animals in retention of the passive-avoidance task. In addition, test results in the animals treated with MK-801 before injury and reinjected with MK-801 before passive-avoidance testing did not differ from those in untreated injured animals reinjected with saline before passive-avoidance testing. These findings indicate that MK-801 treatment before injury prevented the enhanced sensitivity to MK-801-induced amnesia that follows traumatic brain injury.

Topics: Amnesia; Animals; Avoidance Learning; Brain Concussion; Brain Injuries; Dizocilpine Maleate; Male; Rats; Rats, Sprague-Dawley; Reaction Time; Receptors, N-Methyl-D-Aspartate