L. Velíšek

N-Methyl-d-aspartate (NMDA)-induced seizures in developing rats

Intraperitoneal administration of N-methyl-D-aspartate (NMDA) elicited epileptic motor seizures in developing rats aged from 7 to 25 days as well as in young adults. The very first sign of NMDA action is locomotor hyperactivity which is followed by clonic and tonic seizures. In rat pups during the first 3 postnatal weeks flexion seizures (emprosthotonus) appeared as the first pattern of motor seizures; later they were replaced by generalized tonic clonic seizures. Only regular tonic-clonic seizures were observed in 25-day-old and adult rats. The youngest animals are the most sensitive to NMDA. CD50 for tonic-colonic seizures is 6.7 mg/kg in 7-day-old rat pups and it increases up to 86.6 mg/kg in 25-day-old animals. Similar changes could also be demonstrated for LD50, a lethal outcome being very frequent. EEG recordings demonstrated mainly suppressed activity, thus exhibiting a lack of correlation with motor phenomena.

Ketamine suppresses both bicuculline- and picrotoxin-induced generalized tonic-clonic seizures during ontogenesis.

An anticonvulsant action of ketamine, a noncompetitive N-methyl-D-aspartate (NMDA) antagonist (5-40 mg/kg IP), on the bicuculline-induced (3-8 mg/kg IP) or picrotoxin-induced seizures (3-6 mg/kg IP) was assessed in male Wistar rats aged 7, 12, 18, 25 and 90 days. Ketamine alone caused moderate ataxia which was more pronounced in younger animals. In combination with both aforementioned convulsants, ketamine exerted anticonvulsant effects against generalized tonic-clonic seizures in all developmental stages studied. This effect was more pronounced in bicuculline-treated animals. Moreover, ketamine also suppressed the lethality induced by both drugs during all the development. On the contrary, the action of ketamine on minimal (clonic) seizures was moderate or absent. Our results suggest an important role of ketamine-affected transmission in the generation of the generalized tonic-clonic seizure pattern; moreover, an action of high doses of ketamine on GABA-A receptors might be present.

Action of antiepileptic drugs against kainic acid-induced seizures and automatisms during ontogenesis in rats

Summary: Kainic acid (KA 4–14 mg/kg) administered intraperitoneally (i.p.) produces automatisms (scratching until third postnatal week, “wet dog” shakes thereafter), and clonic and tonic‐clonic seizures in rats aged 7, 12, 18, 25, and 90 days. Administration of carbamazepine (CBZ) i.p. (25 or 50 mg/kg), phenobarbital (PB 20–80 mg/kg), clonazepam (CZP 0.2 or 1 mg/kg), or valproate (VPA 200 mg/kg) influenced neither incidence nor latency of automatisms. Clonic seizures that are regularly observed after the third postnatal week in controls were either abolished or substantially suppressed by any of the aforementioned antiepileptic drugs (AEDs). Tonic‐clonic seizures observed in the first 3 postnatal weeks were suppressed only by solvent [including propyleneglycol (PEG), ethanol, and water]; the effect of AEDs on tonic‐clonic seizures was proconvulsant instead. The automatisms were most resistant to AED therapy. These results induce some doubts about the adequacy of the KA model for identifying AEDs effective against complex partial seizures, but forthcoming AEDs that suppress automatisms in the KA rat model might also be active against human complex partial seizures.

Picrotoxin-induced tonic-clonic seizures and lethality are decreased by MK-801 in developing rats

The action of MK-801 (NMDA antagonist; 0.1 and 0.5 mg/kg, IP) was tested against picrotoxin-induced seizures (3-6 mg/kg, IP) in rats aged 7, 12, 18, 25, and 90 days. We found MK-801 only inconsistently affected clonic seizures in 12- and 25-day-old rats, whereas tonic-clonic seizures were suppressed or delayed in almost all age groups. In addition, the lethality of picrotoxin was diminished by the higher dose of MK-801 in all age groups. The results suggest: a) different generators for both seizure patterns (clonic and tonic-clonic), b) an involvement of NMDA receptors in the genesis of tonic-clonic seizure pattern, and c) an interaction of MK-801 with GABAergic transmission throughout the entire development studied.

Kainate/AMPA receptor antagonists are anticonvulsant against the tonic hindlimb component of pentylenetetrazol-induced seizures in developing rats.

Non-NMDA receptor antagonists CNQX, DNQX, and NBQX (10-40 mg/kg IP) were tested against pentylenetetrazol-induced (100 mg/kg SC) seizures in 7 to 90-day-old rats. All three drugs significantly decreased the incidence of tonic hindlimb component of tonic-clonic pentylenetetrazol seizures, often in favor of increased incidence of forelimb tonus throughout development. In addition, in 7 to 25-day-old rats, DNQX and NBQX decreased the severity of seizures due to a decrease in total incidence of the tonic component of tonic-clonic seizures compared to age-matched controls. However, neither drug was able to consistently suppress the incidence or increase latency to onset of clonic and tonic-clonic pentylenetetrazol seizures. The data suggest that, during development, non-NMDA receptor transmission may play a role in the generation of the tonic component, but not in the generation of other components of pentylenetetrazol-induced seizures.

Models of simple partial and absence seizures in freely moving rats: Action of ketamine

The action of ketamine was studied in two models of seizures: a) bilateral neocortical discharges produced by topical application of pentylenetetrazol (model of simple partial seizures); and b) rhythmic spike-and-wave activity induced by systemic administration of pentylenetetrazol (model of absence seizures). Ketamine exerted biphasic effects. In the first model, the dose of 20 mg/kg ketamine significantly suppressed the ictal neocortical discharges (i.e., continuous spiking or ictal activity) accompanied by clonic motor seizures. However, at the dose of 40 mg/kg ketamine significantly accentuated the onset and increased the number of individual discharges (interictal spikes) in bilateral neocortical foci. In the model of rhythmic spike-and-wave activity, the lower dose of ketamine (20 mg/kg) decreased the number of rhythmic spike-and-wave episodes when compared to the higher dose (40 mg/kg) of ketamine, which increased the number of episodes. However, neither result differed significantly from control values. The present results suggest a dose-dependent action of ketamine: Lower doses (10 and 20 mg/kg in the rat) are able to suppress seizure activity, whereas a higher dose (40 mg/kg) potentiates the seizures. Moreover, the action of ketamine may be dependent upon the seizure model used. The study presents a new model of acute epileptic focus in freely moving rats.

Effects of a Benzodiazepine, Bretazenil (Ro 16‐6028), on Rhythmic Metrazol EEG Activity: Comparison with Standard Anticonvulsants

A novel anticonvulsant benzodiazepine breta‐zenil (Ro 16–6028) was studied electrophysiologically in a model of human absence seizures: rhythmic metrazol activity (RMA) in rats. The effects of Ro 16–6028 pretreatment (0.01, 0.05, or 0.1 mg/kg intraperitoneally, i.p.) were compared with those of clonazepam (CZP, 0.02 or 0.1 mg/kg i.p.), valproate (VPA, 200, 300, or 400 mg/kg) and ethosuximide (ESM, 31.25, 62.5, or 125 mg/kg i.p.) in 45 rats with implanted electrocorticographic electrodes. RMA was elicited by an injection of pentylenetetrazol (metrazol, PTZ) in a dose of 40 or 35 mg/kg i.p. The effects of Ro 16–6028 were similar to those of CZP and VPA, i.e., suppression of RMA episodes, an increase in latency and a decrease in number, and total as well as mean duration. On the other hand, ESM differed from these antiepileptic drugs (AEDs) in inability to shorten the duration of RMA episodes. Based on these results, Ro 16–6028 might be predicted to be efficient against human absence seizures.

Excitatory amino acid antagonists and pentylenetetrazol-induced seizures during ontogenesis: III. The action of kynurenic acid and glutamic acid diethylester

N-methyl-D-aspartate (NMDA) receptor antagonists are anticonvulsant drugs with specific activity against tonic-clonic pentylenetetrazol-induced seizures. However, they do not affect clonic seizures with preserved righting reflexes. In these experiments, we tested the anticonvulsant activity of strychnine-insensitive glycine receptor (at the NMDA site) antagonist kynurenic acid and nonspecific excitatory amino acid receptor antagonist glutamic acid diethylester (GDEE) in the pentylenetetrazol-induced seizure model in developing rats 7, 12, 18, 25, and 90 days old. Control rats were injected with pentylenetetrazol (100 mg/kg subcutaneously). Other rats were pretreated either with kynurenic acid (40, 80, or 240 mg/kg IP) or with GDEE (0.48-480 mg/kg IP), followed by pentylenetetrazol (100 mg/kg). In very young rats (7 and 12 days), both kynurenic acid and GDEE increased the incidence of clonic seizures, whereas the occurrence of tonic-clonic seizures was suppressed or delayed compared to controls. This effect is very similar to the anticonvulsant action of the competitive and noncompetitive NMDA receptor antagonists. In adult rats, the pretreatment with rather higher doses of kynurenic acid or GDEE suppressed or delayed clonic seizures as well as tonic-clonic seizures. Both drugs also induced behavioral side effects: repetitive orientation, wet dog shakes, and frequent jumping. Our data show that there are only weak and nonconsistent age-specific anticonvulsant effects resulting from the blockade of strychnine-insensitive glycine receptor often associated with serious side effects, thus decreasing chances to develop effective antiepileptic treatment in this drug class.

Ketamine blocks cortical epileptic afterdischarges but not paired-pulse and frequency potentiation

Cortical epileptiform afterdischarges (spike-and-wave rhythm) induced by low-frequency stimulation of the sensorimotor cortex were dose-dependently shortened by ketamine (10, 20 and 40 mg/kg, i.p.). Myoclonic jerking during stimulation was unaffected by ketamine whereas the same motor pattern accompanying afterdischarges was markedly suppressed by this drug. Paired-pulse as well as frequency potentiation of the cortical interhemispheric (transcallosal) responses were not significantly altered by the 40 mg/kg dose of ketamine. The two simple potentiation phenomena studied probably did not play a role in initiation of cortical epileptic afterdischarges.

Bicuculline-induced neocortical epileptiform foci and the effects of 6-hydroxydopamine in developing rats

Catecholamines (dopamine and norepinephrine) are considered to be predominantly inhibitory neurotransmitters in the brain and their depletion produced by 6-hydroxydopamine may result in proconvulsant effects. In our experiments on rats aged 5, 7, 9, 12, 15, 18, 25 and 90 days under urethane anesthesia we demonstrated the development of neocortical epileptic focus evoked by topical application of bicuculline methiodide. In experimental groups aged 7, 12, 18, 25 and 90 days a chronic depletion of catecholamines was induced using pretreatment with 6-hydroxydopamine early postnatally. An epileptogenic focus was induced in all age groups; duration of a single discharge decreased with age in both control and experimental animals. The spread of activity from the primary focus to contralateral frontal cortex via callosal connections was as rapid as in controls. However, the transfer of discharge to occipital regions was delayed and the number of discharges decreased in experimental rats. Our study demonstrated a substantial role of catecholamines for synchronization of focal discharges in neocortex and a promoting role of catecholamines in association pathways within neocortex.