Development Of Amygdaloid Kindling Research Articles

Stimulation of the superior cerebellar peduncle during the development of amygdaloid kindling in rats

Cerebellar manipulations have been used successfully in some intractable epileptic patients, however, their intrinsic mechanisms are not fully understood. To further clarify the cerebellar participation in epilepsy, we stimulated 10 rats with 100 Hz, 20 μA at the superior cerebellar peduncle (SCP) during amygdaloid kindling. Results were compared to 10 rats with an electrode placed at the SCP without stimulation and 10 rats without electrodes at the SCP used as control. We found that SCP stimulation increased the theta and alpha rhythms at the contralateral motor cortex. Such a stimulation produced hypertonicity of the forelimbs and tremor of the head. In this condition, we found that each of the behavioral stages during amygdaloid kindling in the SCP stimulated rats was reached earlier, while the amygdaloid electrographic afterdischarges (ADs) were longer during the first and shorter in the final trials as compared to controls. Moreover, amygdaloid ADs recorded exclusively during the behavioral stage-5 were significantly shorter than those recorded in the control conditions. We suggest that SCP stimulation could change the customary electrographic and convulsive expression of amygdala kindling in such a manner as to initially facilitate the limbic seizures and impede the secondary generalized seizures.

Involvement of central histamine in amygdaloid kindled seizures in rats

The involvement of central histamine in amygdaloid kindled seizures in rats was investigated using histamine-related compounds. Histamine contents in the amygdala of electrical stimulation site was significantly decreased after development of amygdaloid kindling. Intracerebroventricular (i.c.v.) injection of histamine resulted in inhibition of amygdaloid kindled seizures. The H 1-agonists 2-methylhistamine and 2-thiazolylethylamine also inhibited amygdaloid kindled seizures. In addition, intraperitoneal (i.p.) injection of histidine and metoprine inhibited amygdaloid kindled seizures at doses that caused increases in histamine contents of the brain. H 1-antagonists (diphenhydramine and chlorpheniramine) attenuated histamine (i.c.v.)-induced inhibition of amygdaloid kindled seizures, however, no significant antagonism was observed with H 2-antagonists (cimetidine, ranitidine or zolantidine). Intracerebroventricular injection of H 3-antagonists (thioperamide and AQ 0145) resulted in a dose-related inhibition of amygdaloid kindled seizures. The same findings were observed when thioperamide and clobenpropit were injected i.p. The effects of thioperamide (i.p.) and AQ 0145 (i.p.) were inhibited by an H 3-agonist [(R)-α-methylhistamine] and H 1-antagonists (diphenhydramine and chlorpheniramine). On the other hand, H 2-antagonists (cimetidine and ranitidine) showed no antagonistic effects. These findings suggested that a histaminergic mechanism plays an important role in suppressing amygdaloid kindled seizures through histamine H 1-receptors.

Role of central histamine in amygdaloid kindled seizures

The role of central histamine in amygdaloid kindled seizures in rats was studied. Histamine content in the amygdala was significantly decreased after development of amygdaloid kindling. Intracerebroventricular (i.c.v.) injection of histamine resulted in inhibition of amygdaloid kindled seizures. The H1-agonists 2-methylhistamine and 2-thiazolylethylamine also inhibited amygdaloid kindled seizures. In addition, intraperitoneal (i.p.) injection of histidine and metoprine inhibited amygdaloid kindled seizures at doses that caused increases in histamine contents of the brain. H1-antagonists (diphenhydramine and chlorpheniramine) attenuated histamine- or histidine-induced inhibition of amygdaloid kindled seizures. Both i.c.v. and i.p. injections of H3-antagonists (thioperamide, AQ0145 and clobenpropit) resulted in a dose-related inhibition of amygdaloid kindled seizures. The effects of thioperamide and AQ0145 were inhibited by an H3-agonist (R)-alpha-methylhistamine and H1-antagonists. On the other hand, H2-antagonists showed no antagonistic effect. GABAmimetic drugs, diazepam, sodium valproate and muscimol potentiated the effect of clobenpropit. Bicuculline caused significant antagonism of the inhibition of amygdaloid kindled seizures induced by clobenpropit. These findings suggested that a histaminergic mechanism plays an important role in suppressing amygdaloid kindled seizures through histamine H1-receptors. In addition, an inhibition of amygdaloid kindled seizures induced by histamine is closely related with the action of GABA.

Age-specific effects of noradrenergic alpha-2 agonist clonidine on the development of amygdaloid kindling in developing rats

The effects of clonidine on the development of amygdaloid kindling were studied in rats of various ages (14, 21, 28 and 70 postnatal days). Administration of clonidine (0.2, 0.5 mg/kg i.p.) caused a significant retardation of kindling development in the 28-day-old rats as well as in the adult rats, whereas, in the 14-day-old rats, the development of kindling was significantly facilitated by clonidine. No significant effect of clonidine was observed in the 21-day-old rats. These results indicate that in rats the effects of clonidine on the development of amygdaloid kindling vary during development.

Histaminergic mechanisms in amygdaloid-kindled seizures in rats.

The present study was undertaken to clarify the role of histaminergic neuron system on amygdaloid kindled seizures in rats. A significant decrease in histamine contents in the amygdala was observed after development of amygdaloid kindling. Histidine and metoprine inhibited amygdaloid kindled seizures at doses causing an increase in histamine contents of the brain. H1-antagonists (diphenhydramine and chlorpheniramine) attenuated histidine-induced inhibition of amygdaloid kindled seizures, however no significant antagonism was observed with H2-antagonists (zolantidine and ranitidine). The development of amygdaloid kindling was retarded by repeated administration of histidine. These findings suggest that histaminergic mechanisms play a suppressive role in amygdaloid kindled seizures through histamine H1-receptors.

Specific group II metabotropic glutamate receptor activation inhibits the development of kindled epilepsy in rats

The effects of intracerebral administration of the group II metabotropic glutamate receptor agonist, 2 R,4 R-APDC, were tested on both the development of amygdaloid kindling and on fully developed stage 5 amygdala kindled seizures. The development of amygdaloid kindling was significantly retarded in 2 R,4 R-APDC (10 nmol in 0.5 μl) treated animals compared to control animals over a period of 8 days. At a low dose, 2 R,4 R-APDC (0.1 nmol) caused a 42.5±26.6% increase of the generalised seizure threshold in fully kindled animals. As higher doses were administered, however, the changes in generalised seizure threshold were less marked, and even a small decrease in the threshold was seen (−19.6±5.36% at 10 nmol). The agonist 2 R,4 R-APDC inhibited depolarization-induced release of [ 3H] d-aspartate from cortical synaptosomes with an IC 50 value of 0.29 μM. This effect was maximal at 1 μM, and decreased with dose thereafter. These findings suggest that the selective activation of the group II metabotropic glutamate receptors by agonists such as 2 R,4 R-APDC may be of therapeutic potential in the treatment of seizure disorders.

Role of serotonin receptor subtypes in the development of amygdaloid kindling in rats

The present study was conducted to identify serotonin (5-HT) receptor subtypes involved in the development of amygdala (AM) kindling. We used 8-hydroxy-2-(di- n-propylamino)tetralin (8-OH-DPAT), a 5-HT 1A agonist, and 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), a 5-HT 2 agonist, both of which were injected subcutaneously 15 min prior to each daily electrical stimulation to the rat AM. Treatment with 8-OH-DPAT (1 mg/kg) slightly suppressed behavioral and electrographic seizure development during the course of kindling. In contrast, DOI (1 mg/kg) strongly facilitated kindling development and reduced the number of stimulations needed to produce generalized seizures. These facilitatory effects of DOI were completely blocked by pretreatment with a 5-HT 2 antagonist ketanserin. The present results suggest that the activation of 5-HT 1A receptors can retard the development of AM kindling, whereas 5-HT 2 receptors play a facilitatory role in this developmental seizure process.

Effects of chronic morphine pretreatment on amygdaloid kindling development, postictal seizure and suppression and benzodiazepine receptor binding in rats.

Effects of chronic morphine pretreatment on the development of amygdaloid kindling, seizure suppression and benzodiazepine (BDZ) receptor binding in rats were evaluated. The morphine-pretreated animals showed faster acquisition of seizure activity. Further evaluation of the postictal seizure suppression immediately after a fully kindled seizure demonstrated that morphine-pretreated rats had a decreased sensitivity to subsequent kindling stimulations. Twenty-four hours after the last electrical stimulation, saline-pretreated fully kindled rats showed enhanced BDZ receptor binding in dentate gyrus, and decreased binding in cingulate cortex ipsilateral to the stimulation site, compared to saline controls. Morphine-pretreated amygdala-kindled rats had significantly higher BDZ binding in piriform, entorhinal and sensorimotor cortices, basolateral and cortical amygdaloid nuclei, dentate gyrus, CAI-3 areas, substantia nigra pars reticulata and periaqueductal gray. The present study indicates that the previous experience with chronic morphine modifies the kindling process and that the enhanced BDZ receptor binding detected in our experiments may be involved in the enhanced postictal seizure suppression observed in these animals.

Blockade of both epileptogenesis and glutamate release by (1S,3S)-ACPD, a presynaptic glutamate receptor agonist

The influence of intracerebrally focally administered doses of a presynaptic metabotropic glutamate receptor agonist, (1S,3S)-ACPD, and of the post-synaptically targeted competitive NMDA receptor antagonist, d-CPPene (SDZ EAA 494), was tested on the development of amygdaloid kindling. The actions of these drugs, compared to that of d-CPP, was also tested on fully developed stage 5 amygdala kindled seizures. Both (1S,3S)-ACPD and d-CPPene dose-dependently increased the generalised seizure threshold in fully kindled animals. They showed a similar potency, with (1S,3S)-ACPD acting presynaptically and d-CPPene postsynaptically. Both drugs reversibly inhibited epileptogenesis at 10 nmol in 0.5 μl of injection vehicle, keeping the kindling stage at or below stage 2. All animals reached stage 5 after withdrawal of the 2 drugs. Whereas (1S,3S)-ACPD inhibited depolarisation-induced release of [ 3H] l-glutamate and [ 3H] d-aspartate from cortical synaptosomes (IC 50 63 μM and 50 μM, respectively), d-CPPene (postsynaptically active) was without effect. These findings suggest a new approach to the development of clinically effective anticonvulsants through the development of presynaptic glutamate receptor agonists which could be administered systematically to control the extent of synaptic release of glutamate.

Changes in TRH and its degrading enzyme pyroglutamyl peptidase II, during the development of amygdaloid kindling

Pyroglutamyl peptidase II (PPII) is a neuronal ectoenzyme responsible for thyrotropin releasing hormone (TRH) degradation at the synaptic cleft. PPII, heterogeneously distributed in different brain regions and adenohypophysis, is regulated under various endocrine conditions where TRH is involved in thyrotropin or prolactin regulation but only at the adenohypophyseal level. TRH can downregulate PPII activity in cultured adenohypophyseal cells. TRH present in extrahypothalamic brain areas has been postulated to serve as a neuromodulator and levels of this peptide increase in amygdala, hippocampus and cortex after electrical stimulation (kindling or electroshock). To study whether brain PPII could be regulated in conditions that stimulate TRHergic neurons, TRH and PPII activity were determined during the development of amygdaloid kindling in the rat. TRH levels increased from stage II to V in amygdala and hippocampus in the ipsi- and contralateral side to stimulation. In n. accumbens a decrease, compared to sham was observed at stage II, but levels raised through stage V. In contrast, PPII activity was increased at stage II, in amygdala of both sides and in hippocampus, frontal cortex, n. accumbens and hypothalamus of the contralateral side; levels decreased at stage V to sham values in most structures (except amygdala and hippocampus where the activity was 30% below controls). These results suggest that PPII activity in the central nervous system can be regulated in conditions known to affect TRHergic neurons.

Simultaneous monitoring of the seizure-related changes in extracellular glutamate and γ-aminobutyric acid concentration in bilateral hippocampi following development of amygdaloid kindling

We simultaneously monitored the seizure-related changes in extracellular hippocampal glutamate (Glu) and gamma-aminobutyric acid (GABA) concentration in brain dialysates in order to clarify the role of Glu and GABA in the development of kindling. Brain dialysates were collected every 5 min from 10 min prior to 80 min after stimulus in the three developing conditions consisting of pre-kindling state, stage 3 (C-3), and five consecutive stage 5 (5*C-5) following kindling in the same rat. Extracellular Glu level increased rapidly, lasting for only 5 min after stimulus. The post-stimulus ratio of Glu increase in partially kindled rats (C-3) was 2.5-3.5 times of the baseline, and in fully kindled rats it was about 5 times of the baseline. Extracellular GABA concentration enhanced gradually, reaching a plateau level at 15-20 min and lasting for several hours after stimulus at each stage. The enhancement of GABA level was about 1.5 times of the baseline in partially kindled stage, and was about 2.5 times of the baseline in fully kindled stage. There was no significant difference between the two hemispheres with respect to either the time-course or the magnitude of Glu and GABA increase respectively. These data show that progressive, transient and stimulus-induced enhancement of extracellular Glu levels combined with long-lasting elevation of extracellular GABA levels in the bilateral ventral hippocampi results in imbalance between the excitatory and inhibitory neuronal systems, causing excessive propagation of seizure activity, culminating in the secondary generalized seizure of amygdaloid kindling.

Bilateral seizure-related changes of extracellular glutamate concentration in hippocampi during development of amygdaloid kindling

To monitor the seizure-related changes of extracellular hippocampal glutamate (Glu) concentration during the development of amygdaloid kindling, we used brain dialysates and an enzymatic cycling technique for Glu determination with a highly sensitive assay and high time resolution (1 min). The extracellular Glu level was transiently (for 3 min) enhanced after stimulus and returned rapidly to baseline. In partially kindled rats (stage 3), the extracellular Glu level during the first minute post stimulus was 2.5–3.5-fold that of baseline, while fully kindled rats exhibited about a 5-fold increase in Glu level. Amygdaloid kindling is accompanied by a progressive, transient, stimulus-induced enhancement of extracellular Glu levels during the first minute post stimulus in both hippocampi.

Involvement of Putrescine in the Development of Kindled Seizure in Rats

During the development of kindling by daily electrical stimulations applied to the left amygdala of rats, concentrations of the polyamines putrescine, spermidine, and spermine were measured in the left amygdala and the remainder of the cerebrum. A significant increase of putrescine concentration appeared first at the left amygdala in prekindled rats and then propagated to the remainder of the cerebrum with the development of kindling. This increase in putrescine concentration in the left amygdala was higher in prekindled rats than in fully kindled rats and lasted for at least 24 h after the final kindling stimulation. The concentrations of spermidine and spermine were slightly increased in a fully kindled state. To clarify the role of putrescine in kindling, the development of amygdaloid kindling was examined in rats after microinjections of alpha-difluoromethylornithine, a specific inhibitor of polyamine synthesis, and putrescine into the ipsilateral amygdala. Pretreatment with alpha-difluoromethylornithine (50 nmol) for 10 days accelerated both the development of behavioral kindling and the propagation of the afterdischarge from the left amygdala to the frontal cortex. In contrast, pretreatment with putrescine (200 nmol) for 10 days retarded the development of kindling. These results suggest that the increase in putrescine concentration in the kindled brain has an inhibitory effect on the development of kindling.

Effects of chronic naloxone pretreatment on amygdaloid kindling in rats

Effects of chronic naloxone pretreatment (75 or 270, μg/h for 14 days) on the development of amygdaloid kindling in rats were evaluated. The acquisition of seizure activity was modified in the naloxone pretreated animals, depending on the nucleus stimulated: facilitation of stages IV and V occurred in 37%, variability of electrographic and behavioral responses to electrical stimulation during the kindling development in 33%, and facilitation of stages IV and V followed by long periods of seizure suppression in 29%. Enhancement of postictal seizure suppession during a recycling paradigm was observed in all the naloxone pretreated rats. It was concluded that the chronic administration of naloxone (known to induce opioid binding upregulation and supersensitivity), in association with the enduring changes in opioid mechanisms provoked by kindled seizures, were responsible for the facilitation and suppression of epileptic activity. These findings support bidirectional modulatory effects of opioid peptides on epileptic seizures as well as the view that epileptic seizures can induce enduring alterations in opioid mechanisms.

Amygdaloid kindling during wakefulness and paradoxical sleep in the cat. 1. Inhibitory influence of paradoxical sleep on kindling development

The development of amygdaloid kindling during wakefulness (W) and paradoxical sleep (PS) was compared in chronically implanted adult male cats. The animals were divided into 2 groups. One group was kindled during W (W-K) and the second during a burst of pontogeniculo-occipital (PGO) potentials during PS (PS-K). The threshold of amygdaloid afterdischarge (AD) was determined during W in the W-K group and during PS in the PS-K group. For the development of kindling, amygdala stimulation was applied daily. Results demonstrate that kindling development was markedly retarded in the PS-K group. From the first kindling trials, mean AD duration in the PS-K group was significantly shorter than in the W-K group and this difference was sustained until generalized convulsive seizures (GCSs) were reached by the W-K animals. Also, mean AD frequency in the PS-K group was significantly lower than in the W-K group. AD propagation to the contralateral amygdala and sensorimotor cortex was significantly retarded in the PS-K animals. Time spent in behavioral stages I and II of kindling was significantly longer in the PS-K animals than in W-K animals and the number of daily electrical stimuli required to reach the first GCS was significantly higher in group PS-K than in group W-K. It is concluded that PS exhibits an inhibitory influence over amygdaloid kindling development and this influence is mainly exerted during the early stages of epileptogenesis.

Intraventricular injection of vasoactive intestinal polypeptide facilitates the development of amygdaloid kindling

To determine the involvement of vasoactive intestinal polypeptide (VIP) in the development of kindling in rats, VIP or antibody to VIP (anti-VIP) was injected into the third ventricle after the daily stimulation of the basolateral amygdala. The number of stimulations required to reach full amygdaloid kindling was significantly smaller in rats treated with VIP than rats treated with saline, whereas it increased significantly in rats treated with anti-VIP compared to that in the rats treated with either saline or normal rabbit serum. The results suggest that VIP participates in the development of amygdaloid kindling as a facilitatory factor.

Time-dependent pro- and anticonvulsant effects of cysteamine on the development and expression of amygdaloid kindled seizures

The time-dependent pro- and anticonvulsant effects of cysteamine, a depletor of somatostatin, were investigated on the development and expression of amygdaloid kindled seizures. Acute administration of cysteamine (25–400 mg/kg, i.p.) produced a dose-dependent potentiation of kindled seizures when evoked 4 h after the drug. However, the seizures initiated 1 day after drug administration were dose-dependently suppressed. Furthermore, elicitation of seizures 4 h after cysteamine enhanced its anticonvulsant effects at 1 day after the drug, causing a parallel left shift of the dose-responsecurve. Since it has been reported that somatostatin is released during generalized seizures, the seizures given 4 h after cysteamine may encourage the somatostatin depletion by cysteamine and thereby potentiate its later anticonvulsant effects. The repeated administration of cysteamine (100 mg/kg, i.p.) during kindling development strongly retarded the development of generalized seizures but not the development of focal seizures or of afterdischarges in the amygdala. In contrast to the acute experiments, kindling stimulation given 4 h after each cysteamine treatment did not augment the blocking effect on kindling development. These data indicate that chronic cysteamine treatment has a strong inhibitory effect on the development of amygdaloid kindling.

Changes in the dynamic state of brain proenkephalin-derived peptides during amygdaloid kindling.

The dynamic state of the proenkephalin (PE) gene products during and after development of amygdaloid kindling was assayed by monitoring changes of the accumulation of PE mRNA and changes in proenkephalin-related peptides. A parallel determination of PE mRNA and peptides from the same sample was conducted in this study. Electrical stimulation of the amygdala causes early increases in the PE mRNA content in that structure and in the hippocampus. Other areas related with the amygdaloid complex do not exhibit such an early increase, but this alteration occurs when the kindling process is fully established. Enkephalin content increases early in amygdala and hippocampus presumably owing to an increase in synthesis rate. Also, the enkephalin content of areas connected with amygdala and hippocampus such as the entorhinal cortex, the nucleus accumbens, and the frontal and occipital cortex exhibits an increase. A clear tendency towards normalization is observed after a recovery period of 2-3 months. Rekindling of the animals after this recovery period does not elicit a similar pattern of changes in the dynamic state of enkephalin system, even though the animals rekindle with just one single stimulation. The present data suggest that the enkephalinergic neurons participate in the development and spreading of kindling phenomena after amygdaloid stimulation, but they do not seem to play any role in mediating maintenance of the kindling state.

Amygdaloid kindling during rapid eye movement (REM) sleep in cats

Development of amygdaloid kindling was analyzed during REM sleep and during wakefulness. Daily evolution of electrographic and behavioral changes was significantly delayed in REM kindled rats. The number of kindling trials required to reach the first generalized convulsive seizure was also significantly increased in comparison with awake kindled animals. Changes in sleep organization were measured under REM kindling conditions. A significant increase in total sleep time and in the percentage of light slow-wave sleep was found during the kindling process. No significant sleep changes were observed in REM-established kindling. REM inhibitory influence over epileptogenesis is discussed.

Effet de la dl-allylglycine sur l'embrasement amygdalien chez le rat

The effect of decreased cerebral GABA levels on the development of amygdaloid kindling in rats was investigated. DL-Allylglycine, an inhibitor of glutamic acid decarboxylase (GAD), administered for 2 or 9 days (100 mg/kg and 50 mg/kg respectively) was found to accelerate the kindling process. The results are discussed according to a possible role of GABA mechanisms in the kindling phenomenon.