Lack of effect of secretin on kindling and seizures

Secretin infused into rats activates neurons located in brain areas controlling autonomic function and emotion. The brain activity of secretin is mediated, at least in part, through vagal pathways. It is known that afferent stimulation of the vagus nerve results in considerable antiepileptic effects. Whether or not secretin has an effect on seizures is unknown. In this study, we evaluated the efficacy and safety of secretin as an antiepileptogenic agent in electrical kindling and as an anticonvulsant in fully kindled seizures. To assess antiepileptogenic effects, we administered secretin (10, 30, or 100 microg/kg/dose) or normal saline intravenously 5 min before twice-daily kindling stimulation. To assess the anticonvulsant effect of secretin, we administered either normal saline or secretin (100 microg/kg/dose) 5 min before the electrical stimulation to fully kindled rats. We observed no effect on kindling rate or afterdischarge duration. In fully kindled rats, secretin administration had no effect on kindling stage or afterdischarge duration. Thus, in the dose range used in this preliminary acute treatment study, secretin had no discernible antiepileptogenic or anticonvulsant effects. Secretin was very well tolerated in this multidose protocol.     

The effect of repeated seizures on anticonvulsant drug response in the kindling model

Clinical reports suggest that the occurrence of numerous seizures prior to the onset of anticonvulsant therapy is associated with a poor prognosis for drug control. This seems to imply that seizures become harder to suppress as they recur--a possibility which argues for the early initiation of anticonvulsant therapy. The present study attempted to determine whether this effect could be demonstrated under controlled laboratory conditions. The kindling preparation, an animal model involving the elicitation of repeated focal seizures, was used. Drug response (ED50) was measured in Royal Victoria hooded rats after either a small or a large number of pretreatment seizures, administered in the kindling paradigm. A variety of experiments were performed, involving: different drugs (phenobarbital, phenytoin, and carbamazepine), different seizure types (amygdala focal seizure, cortical focal seizure, generalized convulsion), and different stimulation parameters (0, 20, 40 or 100 pretreatment seizures: 'near-threshold' vs. 'standard' stimulation). In no case were seizures found to be harder to suppress following repeated pretreatment seizures. After large numbers of pretreatment seizures (40, 100), drug response was actually enhanced. These data indicate that the mere repetition of seizures does not automatically lead to a decrease in anticonvulsant effectiveness. They offer no particular rationale for the early initiation of anticonvulsant therapy in clinical situations.     

The effect of minocycline on seizures induced by amygdala kindling in rats

PurposeMinocycline is known as a chemical with neuroprotective, anti-inflammatory, and antimicrobial properties. In this study, the effects of minocycline on seizures induced by amygdala kindling in rats were studied.MethodsKindled Wistar rats were injected intraperitoneally with saline and, on the following day, with minocycline (50, 25, and 12.5 mg/kg for the three groups (1–3), respectively). The animals in groups 1–3 had similar protocols. Groups 4 and 5 were given for the rotarod test and received 25 or 50 mg/kg minocycline, respectively, without any kindling stimulation. The animals in groups 6 and 7 (seven each) received 25 mg/kg minocycline or saline, respectively. All the injections were carried out 1 h before kindling stimulation. Seizure parameters, including after discharge duration (ADD), stage 4 latency (S₄L), stage 5 duration (S₅D), and seizure duration (SD), were recorded and compared with those of the saline groups.ResultsMinocycline (50 mg/kg) significantly reduced ADD, 1/S₄L, S₅D, and SD (P < 0.001, P < 0.05, P < 0.001, and P < 0.001, respectively) in group 1. While the administration of 25 mg/kg of minocycline decreased the ADD and S₅D (P < 0.05), in group 2. The injection of 12.5 mg/kg resulted in decreased S₅D (P < 0.001) in group 3. The daily injection of minocycline (25 mg/kg) significantly decreased ADD, S₅D, and SD (P < 0.001) in group 6.ConclusionThe obtained results revealed that minocycline has anticonvulsant effect on seizures induced by amygdala kindling. Thus, it may be useful for epilepsy treatment.     

The effect of glutaurine on thyroid hormones in the rat

Glutaurine (gamma-L-glutamyl taurine), a proposed hormone isolated from parathyroid gland oxyphil cells, was examined for its effect on circulating thyroid hormones in the rat. In acute experiments performed over a 24 hr. period, glutaurine depressed plasma triiodothyronine (T3) levels in a dose-dependent manner; however, thyroxine (T4) levels were not affected significantly. In chronic experiments performed over a 2 wk. period, glutaurine significantly increased T3 levels, but, as with acute studies, the effect of T4 levels was not significantly altered. Following acute glutaurine administration, TSH levels were elevated above control. The increased T3 observed following chronic glutaurine administration may be due to a secondary increase in TSH levels. These data support the hypothesis that glutaurine aids in peripheral thyroid hormonal regulation. Observed differences between acute and chronic glutaurine action are though to result from the effect of glutaurine on the negative feedback inhibiting action of TSH.     

Influences of pituitary-adrenal hormones on kindling

The kindling process represents a progressively augmenting electrical and behavioral response to brain stimulation, that has been proposed as a model for the development of epileptogenesis. The first experiment examined the effects of hypophysectomy on the kindling process. The effects of hypophysectomy were found to be dependent on the number of days between hypophysectomy and the beginning of kindling. The kindling rate was slowed, compared to intact controls, when hypophysectomy was performed less than 2 weeks prior to the first kindling stimulation. However, if more than 4 weeks and passed since hypophysectomy, the kindling rate was faster than that of intact controls. The second experiment tested the influence of systemic administration of cortisone (10 mg/animal), upon kindling of control and hypophysectomized animals. Cortisone was found to reverse the kindling deficit of hypophysectomized animals at less than 2 weeks and to slow kindling of intact controls, but to have no effect on hypophysectomized animals at more than 4 weeks (at this dose). These results are interpreted as indicating that pituitary-adrenal hormones have profound influences on the kindling process, but it is unclear whether the influence is upon brain excitability or more directly upon modification of synaptic function.     

The effect of tricyclic antidepressants on cortex- and amygdala-kindled seizures in the rat

The anticonvulsant effects of amitriptyline, imipramine, nortriptyline and desipramine were tested against focal and generalized seizures, triggered from either the amygdala or the cortex in fully kindled rats. Tests were administered on a 72- or a 24-hour schedule. Significant seizure suppression was achieved with only one drug, amitriptyline, and it occurred only at toxic or near-toxic doses. The differential, low-dose suppression of amygdala-kindled seizures, reported in earlier studies, was not seen in the present experiments. It may occur only in the short-interval test paradigms used by previous experimenters.     

Seizure susceptibility in aged rats--pentylenetetrazol-induced seizures and amygdaloid kindling seizures]

Seizure susceptibility in male aged rats (21-25 mo. age) was investigated by pentylenetetrazol (PTZ)-induced seizures and amygdaloid kindling seizures. Male adult rats (3-4 mo. of age) were used as the control group. During 60 min. after the administration of PTZ (70 mg/kg, s.c.), the aged rats had a higher incidence and a significantly longer duration of generalized convulsion than the control group. On the other hand, the kindling rate of the aged rats was significantly slower than that of the control. The aged rats remained longer at stage 1-2 indicative of partial seizures. However, the aged rats skipped stage 3 on their way to stage 4-5 (generalized seizures), and showed a more violent stage 5 seizure than the control group. They had a longer duration of afterdischarges (AD) through the kindling process with a significantly faster propagation of AD to the contralateral amygdala as compared with the control group. The present study suggests that aged rats are prone to convulsion, while they have a difficulty in acquisition of epileptogenesis.     

Neuronal injury and brain-derived neurotrophic factor expression in a rat model of amygdala kindling seizures Differences in brain regions and kindling courses

BACKGROUND:Studies have demonstrated that brain-derived neurotrophic factor (BDNF) has a dual effect on epilepsy. However, the relationship between epilepsy-induced brain injury and BDNF remains poorly understood.OBJECTIVE:According to ultrastructural and molecular parameters, to detect the degree of neuronal injury and BDNF expression changes at different brain regions and different kindling times to determine the effects of BDNF on epilepsy-induced brain injury.DESIGN, TIME AND SETTING:A randomized, controlled, animal experiment based on neuropathology and molecular biology was performed at the Department of Physiology and Department of Pathology, Basic Medical College of Jilin University in 2003.MATERIALS:UltraSensitiveTM SP kit for immunohistochemistry (Fuzhou Maxim Biotechnology, China), BDNF antibody (concentrated type, Wuhan Boster Biological Technology, China), JEM-1000SX transmission electron microscopy (JEOL, Japan), and BH-2 light microscope (Olympus, Japan) were used in the present study.METHODS:Wistar rats were randomly assigned to control (n = 6), sham-surgery (n = 6), and model (n = 60) groups. The control group rats were not treated; an electrode was embedded into the amygdala in rats from the sham-surgery and model groups; an amygdala kindling epilepsy model was established in the model group.MAIN OUTCOME MEASURES:Pathological changes in the temporal lobe and hippocampus were observed by light and electron microscopy at 1, 3, 7, 14, and 21 days following kindling, and BDNF expression in the various brain regions was determined by immunohistochemistry.RESULTS:In the model group, temporal lobe cortical and hippocampal neurons were swollen and the nuclei were laterally deviated. There were also some apoptotic neurons 3 days after kindling. The nucleoli disappeared and the nuclei appeared broken or lysed, as well as slight microglia hyperplasia, at 7 days. Electron microscopic observation displayed chromatin aggregation in the nuclei and slight mitochondrion swelling 3 days after kindling. Injury changes were aggravated at 7 days, characterized by broken cytoplasmic membrane and pyknosis. With the development of seizure, the number of BDNF-positive neurons in the hippocampus and temporal lobe increased and peaked at 7 days. Moreover, hippocampal and cortical temporal lobe injury continued. Following termination of electrical stimulation after 7 days of kindling, BDNF expression decreased, but continued to be expressed, up to 21 days of kindling. In addition, the number of temporal and hippocampal BDNF-positive neurons was greater than the control group.CONCLUSION:Brain injury and BDNF expression peaked at 7 days after kindling, and hippocampal changes were significant.     

Effects of hypoglycemia on kindling seizures in suckling rats

The effects of insulin-induced hypoglycemia on epileptic disorders of suckling rats were examined using an amygdala kindling model. Kindling stimulations were conducted at 16 and 17 days of age with electrodes implanted in the amygdala 2 days earlier. In 18-day-old kindled rats, which acquired generalized behavioral seizures (stages 4 and 5; Moshé's score) by kindling stimulations, the duration of afterdischarge and behavioral seizures evoked by the stimulation at a threshold intensity to produce a generalized seizure was significantly prolonged after an injection of insulin (25 U/kg, i.p.). The prolongation was not observed in kindled rats that exhibited normal blood glucose concentrations after the application of saline or insulin together with glucose. There were no apparent changes in the amplitude of the afterdischarge, the score of behavioral seizure stages, or the generalized seizure threshold. A similar, marked prolongation of afterdischarge and behavioral seizures following the application of insulin, as in the kindled rats, was also observed during the course of the kindling acquisition without accelerating the development of kindling seizure scores. These results indicate that insulin-induced hypoglycemia easily increases the risk of prolonged seizures in immature brain without precipitating the secondarily generalizing mechanism.     

Effect of serial seizures on subsequent kindling in the immature brain

The hypothesis that seizures permanently alter the mammalian brain, making it more susceptible to further seizures was tested in the immature rat using the kindling model. Rate of kindling and final kindling stage reached was compared in 30-day-old rats previously subjected to 4 daily electroconvulsive seizures and weight-matched littermate controls. There were no significant differences between the 2 groups. This study does not support the hypothesis that seizures increase susceptibility for further seizures.     

Ornithine decarboxylase induction and polyamine synthesis in the kindling of seizures: the effect of alpha-difluoromethylornithine.

It has been suggested that the kindling of seizures may depend on the induction of genes encoding enzymes involved in neurotransmission. Experimental seizures are followed by an especially rapid and massive induction of brain ornithine decarboxylase (ODC), an enzyme which catalyses the rate-limiting step in the synthesis of polyamines. The latter compounds have been shown to act as positive allosteric modulators of the NMDA receptor, and also to play an important role in cell growth and differentiation. The induction of ODC by seizures has accordingly been suggested to play a pivotal role in the changes in synaptic structure and function that underlie kindling. In the present study we examined the progress of kindling during treatment with alpha-difluoromethylornithine (DFMO), an irreversible inhibitor of ODC. We found that progressive increase in the duration and severity of kindled seizures and in the duration of local afterdischarges was unaffected by daily injections of DFMO in doses previously shown to cause substantial depression of brain ODC activity. Treatment with DFMO also failed to produce significant anticonvulsant or proconvulsant effects. Progressive increase in seizure activity during kindling is therefore unlikely to depend to any appreciable extent on enhanced synthesis of polyamines by ODC.     

Development of kindling and spontaneous seizures after massed stimulation of different loci in the rat piriform cortex

Massed electrical stimulation of the anterior piriform cortex (PC) in rats using short (5 min) interstimulus intervals has previously been reported to induce severe chronic epilepsy with spontaneous seizures and has thus proposed to represent a novel model of temporal lobe epilepsy. In the present study, we used this stimulation protocol to evaluate the frequency and severity of recurrent spontaneous seizures produced in this way. In addition to the locus in the anterior PC previously used for massed stimulation (MS), we also stimulated rats via a locus in the transition zone between anterior and posterior PC ("central PC"), which previously was found to be more sensitive to electrical stimulation than various other loci in the anterior or posterior PC. During MS (71 stimulations for 1 s each at twice afterdischarge threshold), focal and infrequent secondary generalized seizures occurred in both groups, but there was no consistent progressive increase in seizure severity with increasing number of seizures, possibly as a result of postictal inhibitory processes. Following MS, rats were restimulated after 1, 2, 4, and 7 weeks, using five stimuli at 5-min interstimulus periods at each retest period. In both PC-implanted groups, seizure severity and seizure duration progressively increased over the period of the retests, indicating a delayed development of kindling. Spontaneous seizures were only observed rarely, so that MS of the PC is certainly no effective means of producing recurrent spontaneous seizures.     

Partial and generalized seizures affect reproductive physiology differentially in the male rat

PURPOSE: Reproductive dysfunction and endocrine disorders occur frequently among men with epilepsy. This study tested the hypothesis that focal limbic seizures and generalized seizures may both contribute to reproductive dysfunction. METHODS: The rat kindling model was used to mimic focal limbic seizures. Kindling electrodes were placed in the basolateral amygdala. Male rats were either intact, gonadectomized (GDX) or GDX + testosterone (T) replaced and then kindled. Controls were left intact and sham-kindled. Maximal electroconvulsive shock (MES) treatment was used to model generalized seizures, by using eight stimulations, one every other day, for 2.5 weeks. Animals were killed either 3 h or 6 weeks after MES treatment to determine short- and long-term effects. RESULTS: Kindled seizures resulted in an increase in serum testosterone, estradiol, and prolactin in intact males, accompanied by a significant increase in testis, epididymis, and pituitary weight, as well as a significant decrease in prostate weight. MES treatment caused a short-term reduction in serum testosterone and testis, epididymis, and prostate weight. All parameters were restored to control values within 6 weeks of the last MES seizure, with the exception of pituitary weight and serum prolactin, which remained significantly elevated 6 weeks after MES treatment. CONCLUSIONS: Our results indicate that both focal limbic (amygdaloid) seizures and generalized MES seizures disturb normal reproductive physiology in the male rat. Amygdaloid-kindled seizures have mixed effects on different parameters of reproductive function, whereas MES seizures induce a transient hypogonadal state. These results suggest that reproductive dysfunction in men with epilepsy may result from seizure-related interference with the normal functions of the hypothalamic-pituitary-testicular axis.     

Altered activities of rat brain metabolic enzymes caused by pentylenetetrazol kindling and pentylenetetrazol--induced seizures

The aim of our study was to investigate amino acid and energy metabolism of pentylenetetrazol (PTZ)-kindled animals. Glutamate dehydrogenase, aspartate-aminotransferase (AST), alanine-aminotransferase, gamma-glutamyltransferase, alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and creatine kinase (CK) were determined in the frontal cortex, cerebellum, hippocampus and pons-medulla regions of Hannover-Wistar rats. The rats were randomly divided into four experimental groups: (a) control; (b) rats which received a single PTZ injection in a subconvulsive dose of 40 mg/kg i.p.; (c) rats which received a single PTZ injection in a convulsive dose of 50 mg/kg i.p.; and (d) PTZ-kindled rats. Kindling increased ALP activity throughout the brain, elevated AST as well as LDH activity in the frontal cortex and hippocampus and decreased CK activity in the frontal cortex and cerebellum. Acute seizures of the same intensity did not induce these alterations. The observed effects therefore are obviously linked to the kindling phenomenon and not to seizure activity. Changes appeared mainly in the frontal cortex and hippocampus, i.e. brain areas believed to be directly involved in kindling.     

The effects of right and left amygdala kindling on the female reproductive system in rats

Purpose:   Women with epilepsy often have comorbid reproductive dysfunction. Using the amygdala kindling model in rats, the present study examined the effects of seizures of limbic origin on the reproductive system.
Methods:   Female Wistar rats were kindled from the left or right basolateral amygdala to a criterion of 40 stage V seizures. Sham-kindled subjects were handled but not stimulated. Vaginal cytology was assessed daily for the duration of the study. Twenty-four hours following the last kindled seizure, kindled subjects and their yoked controls were sacrificed and their brains and serum were extracted.
Results:   Kindled subjects displayed significantly more abnormal estrous cycle days and significantly elevated levels of estradiol as compared to controls. There was, however, no total suppression of cycling. No laterality effects were seen for estrous cycle abnormalities.
Discussion:   Seizures of limbic origin cause changes in estrous cycling. Right and left kindling seem to have a similar effect. These findings highlight the need for clinicians to monitor reproductive issues among individuals with epilepsy.     

Unidirectional interaction between flurothyl seizures and amygdala kindling

In this report, the interaction between flurothyl convulsions and electrical kindling of the amygdala was investigated. Three flurothyl convulsions decreased the afterdischarge threshold of the amygdala and enhanced the rate of development of electrical kindling without affecting the intensity of postictal refractoriness. On the other hand, 3 generalized kindled convulsions did not alter the flurothyl convulsive threshold. The data suggest that the influence of generalized convulsions on future seizure susceptibility may depend on the agents used to induce the convulsions.     

Effect of sexual hormones on the electroencephalogram and seizures

The role of hormones and reproductive function in electroencephalography (EEG) and epilepsy has long intrigued electroencephalographers. In this report, the relationship between the menstrual cycle and pregnancy and EEG changes and seizure frequency is reviewed. These neurophysiologic changes are likely secondary to hormonal effects. The effect of these hormones on experimental animal models is also reviewed.     

Relationships between triggered seizures, spontaneous seizures, and interictal spiking in the kindling model of epilepsy

Cats were kindled in the amygdala. After completion of kindling, their EEG was monitored almost continuously by a computer system which allowed the detection of spontaneous seizures and the quantification of interictal spikes. The relationships between seizures triggered by stimulation, spontaneous seizures, and interictal spikes in kindled and contralateral amygdalas were examined. Very few spontaneous seizures were observed; their occurrence appeared facilitated by a few triggered seizures after a long seizure-free interval. Stimulation-triggered seizures were followed first by a decrease in spiking activity during a few hours and then by an increase which could take 3 to 8 days to return to baseline. Spontaneous seizures were also, but less systematically, followed by such a sequence. Spontaneous seizures could occur when the interictal spiking rate was high as well as when it was low. It is concluded that, in the fully kindled cat, interictal spiking appears to be mainly regulated by the occurrence of seizures; the rate of spiking appears to have no bearing on the probability of occurrence of spontaneous seizures. Hence, seizures and interictal spiking could be generated by separate pathophysiologic mechanisms, the seizure-generating mechanism influencing the spike-generating mechanism but not the reverse.     

The effects of atropine and reserpine on cortical kindling in the rat.

The effects on cortical kindling of atropine (a muscarinic, cholinergic blocking agent) and reserpine (a depleter of catecholamines and 5 hydroxytryptamine) were tested in this study. Atropine, which had previously been found to retard amygdaloid kindling, had similar but somewhat weaker effects on cortical kindling. Reserpine also had similar effects on cortical kindling compared to subcortical kindling in that it potentiated seizure responses.     

Amygdala kindling and anxiety in the rat

In humans, limbic epilepsy seems to predispose to anxiety. Attempts to model this phenomenon have shown that limbic kindling increases anxiety in domestic cats. No comparable data exist in rodents. The present study was done to investigate the effects of unilateral medial amygdala kindling in Wistar rats on behaviour in the 'elevated plus maze' test of anxiety. It was found that kindling to stage 5 seizures increased anxious response in the plus maze for at least a week following the last seizure. There were equally long lasting decreases in exploratory motivation in the hole board test, which were unrelated to the change in anxiety in the plus maze. The relevance of these findings to epilepsy, stress and anxiety are discussed.