Pentylenetetrazol In Rats Research Articles

Diaphragm relaxation causes seizure-related apnoeas in chronic and acute seizure models in rats

Ictal central apnoea is a feature of focal temporal seizures. It is implicated as a risk factor for sudden unexpected death in epilepsy (SUDEP). Here we study seizure-related apnoeas in two different models of experimental seizures, one chronic and one acute, in adult genetically-unmodified rats, to determine mechanisms of seizure-related apnoeas. Under general anaesthesia rats receive sensors for nasal temperature, hippocampal and/or neocortical potentials, and ECG or EMG for subsequent tethered video-telemetry. Tetanus neurotoxin (TeNT), injected into hippocampus during surgery, induces a chronic epileptic focus. Other implanted rats receive intraperitoneal pentylenetetrazol (PTZ) to evoke acute seizures. In chronically epileptic rats, convulsive seizures cause apnoeas (9.9 ± 5.3 s; 331 of 730 convulsive seizures in 15 rats), associated with bradyarrhythmias. Absence of EEG and ECG biomarkers exclude obstructive apnoeas. All eight TeNT-rats with diaphragm EMG have apnoeas with no evidence of obstruction, and have apnoea EMGs significantly closer to expiratory relaxation than inspiratory contraction during pre-apnoeic respiration, which we term “atonic diaphragm”. Consistent with atonic diaphragm is that the pre-apnoeic nasal airflow is expiration, as it is in human ictal central apnoea. Two cases of rat sudden death occur. One, with telemetry to the end, reveals a lethal apnoea, the other only has video during the final days, which reveals cessation of breathing shortly after the last clonic epileptic movement. Telemetry following acute systemic PTZ reveals repeated seizures and seizure-related apnoeas, culminating in lethal apnoeas; ictal apnoeas are central – in 8 of 35 cases diaphragms initially contract tonically for 8.5 ± 15.0 s before relaxing, in the 27 remaining cases diaphragms are atonic throughout apnoeas. All terminal apnoeas are atonic. Differences in types of apnoea due to systemic PTZ in rats (mainly atonic) and mice (tonic) are likely species-specific. Certain genetic mouse models have apnoeas caused by tonic contraction, potentially due to expression of epileptogenic mutations throughout the brain, including in respiratory centres, in contrast with acquired focal epilepsies. We conclude that ictal apnoeas in the rat TeNT model result from atonic diaphragms. Relaxed diaphragms could be particularly helpful for therapeutic stimulation of the diaphragm to help restore respiration.

Brain connectivity analysis in preictal phases of seizure induced by pentylenetetrazol in rats

Abnormal patterns of brain connectivity characterize epilepsy. However, little is known about these patterns during the stages preceding a seizure induced by pentylenetetrazol (PTZ). To investigate brain connectivity in male Wistar rats during the preictal phase of PTZ-induced seizures (60 mg/kg), we recorded local field potentials in the primary motor (M1) cortex, the ventral anterior (VA) nucleus of the thalamus, the hippocampal CA1 area, and the dentate gyrus (DG) during the baseline period and after PTZ administration. While there were no changes in power density between the baseline and preictal periods, we observed an increase in directional functional connectivity in theta from the hippocampal formation to M1 and VA, as well as in middle gamma from DG to CA1 and from CA1 to M1, and also in slow gamma from M1 to CA1. These findings are supported by increased phase coherence between DG-M1 in theta and CA1-M1 in middle gamma, as well as enhanced phase-amplitude coupling of delta-middle gamma in M1 and delta-fast gamma in CA1. Interestingly, we also noted a slight decrease in phase synchrony between CA1 and VA in slow gamma. Together, these results demonstrate increased functional connectivity between brain regions during the PTZ-induced preictal period, with this increase being particularly driven by the hippocampal formation.

Sıçanlarda Pentilentetrazol ile İndüklenen Akut Epilepsi Modelinde Valproik Asidin NO/cGMP Üzerindeki Etkileri

Epilepsy is a disease which causes neuronal damage and loss of consciousness in consequence of recurrent seizures. Nitric oxide as a neuromodulator in brain is a gas which can penetrate into cells. It has a significant role on physiological cases,pathology of many diseases such as inflammation and degenerative diseases. The purpose of this research is to search the activity of NO/cGMP pathway of valproic acid in an experimental acuteepileptic model which is induced with pentylenetetrazol in rats.18 adult male Wistar Albino rats were used in the study. The rats were randomly divided into 3 groups (n=6) as control group, pentylenetetrazol (PTZ+salin) 45 mg kg-1, valproic acid (PTZ+VPA) 150 mg kg-1. After 24 hours of PTZ application, all rats brain tissues were removed and then cortex and hippocampus were separated. While PTZ increased the hippocampus and cortex NO/cGMP levels compared to control (p <0.01), VPA decreased the hippocampusand cortex NO levels by comparison with PTZ (p <0.001). On the other hand, while VPA decreased cortex cGMP levels (p<0,05) it did not change cGMP levels in hippocampus (p> 0.05). This study has suggested that VPA can show antiepileptic activity via NO/cGMP pathway.

Selenium Nanoparticles with Prodigiosin Rescue Hippocampal Damage Associated with Epileptic Seizures Induced by Pentylenetetrazole in Rats

Simple SummaryEpilepsy is a chronic neurological disease characterized by neuronal hyper electrical activity and the development of unprovoked seizures. Although several antiepileptic drugs are currently available, their application is associated with undesirable adverse effects. In an attempt to find a novel antiepileptic medication with minimum side effects, we have investigated the potential neuroprotective activity of prodigiosin, a red pigment produced by bacterial species that have important pharmaceutical and biological activities biosynthesized with selenium formulation (SeNPs-PDG) against a murine epileptic model mediated by pentylenetetrazole. The main recorded findings revealed that SeNPs-PDG delayed the onset of epileptic seizures and decreased their duration significantly. Additionally, SeNPs-PDG prevented hippocampal cell loss, oxidative stress, neuroinflammation, restored the balance between excitatory and inhibitory neurotransmitters, and notably normalized the monoaminergic and cholinergic transmission. These promising findings indicate that SeNPs-PDG might serve as a naturally derived anticonvulsant agent due to their active antioxidant, anti-inflammatory, anti-apoptotic, and neuromodulatory properties.Background: Prodigiosin (PDG) is a red pigment synthesized by bacterial species with important pharmaceutical and biological activities. Here, we investigated the neuroprotective and anticonvulsant activities of green biosynthesized selenium formulations with PDG (SeNPs-PDG) versus pentylenetetrazole (PTZ)-induced epileptic seizures. Methods: Rats were assigned into six experimental groups: control; PTZ (60 mg/kg, epileptic model); sodium valproate (200 mg/kg) + PTZ; PDG (300 mg/kg) + PTZ; sodium selenite (0.5 mg/kg) + PTZ; and SeNPs-PDG (0.5 mg/kg) + PTZ. The treatment duration is extended to 28 days. Results: SeNPs-PDG pre-treatment delayed seizures onset and reduced duration upon PTZ injection. Additionally, SeNPs-PDG enhanced the antioxidant capacity of hippocampal tissue by activating the expression of nuclear factor erythroid 2–related factor 2 and innate antioxidants (glutathione and glutathione derivatives, in addition to superoxide dismutase and catalase) and decreasing the levels of pro-oxidants (lipoperoxidation products and nitric oxide). SeNPs-PDG administration inhibited inflammatory reactions associated with epileptic seizure development by suppressing the production and activity of glial fibrillary acidic protein and pro-inflammatory mediators, including interleukin-1 beta, tumor necrosis factor-alpha, cyclooxygenase-2, inducible nitric oxide synthase, and nuclear factor kappa B. Moreover, SeNPs-PDG protected against hippocampal cell loss following PTZ injection by decreasing the levels of cytosolic cytochrome c, Bax, and caspase-3 and enhancing the expression of anti-apoptotic Bcl-2. Interestingly, SeNPs-PDG restored the PTZ-induced imbalance between excitatory and inhibitory amino acids and improved monoaminergic and cholinergic transmission. Conclusions: These promising antioxidative, anti-inflammatory, anti-apoptotic, and neuromodulatory activities indicate that SeNPs-PDG might serve as a naturally derived anticonvulsant agent.

Mild foot electrical stimulation is comparable with phenytoin in inhibiting pentylenetetrazol-induced kindling in rats.

Increasing evidence demonstrates that electric stimulation has anticonvulsant effects. The present study was undertaken to investigate the effects of mild foot electrical stimulation (MFES) on the development of pentylenetetrazol (PTZ) kindling and compare its effectiveness with the more commonly used treatment, phenytoin. Kindling was induced in rats by repeated injections (every 24h) of PTZ (37.5mg/kg). The rats were subjected to either MFES (0.2mA in intensity for a 160ms duration with a 160ms interval for 20min) or phenytoin (30mg/kg) before PTZ injections. Following this treatment, rats received MFES every other day for 10days or 26days after establishment of PTZ kindling. The data showed that MFES significantly inhibited development of chemical kindling induced by PTZ in rats (p = 0.001, as compared to PTZ-treated animals). This inhibitory effect is comparable with the effect of 30mg/kg doses of phenytoin (P = 0.99, as compared to phenytoin group). However, 10days or 26days durations of MFES had no effect on established kindled seizures (P = 0.58 as compared to PTZ-treated animals). Our data demonstrate that although MFES significantly inhibited the development of chemical kindling, this experimental paradigm had no effect on established kindled seizures.

Investigating the effect of rTMS on acute pentylenetetrazol injection induced seizures in rat

Absence epilepsy is a kind of epileptic syndrom which has not an absolute treatment. So much research to find new ways to treat the disease is in progress. On the other hand, it has been shown that application of repetitive transcranial magnetic stimulation (rTMS) can be applied through magnetic induction causes changes in neuronal excitability. In the present study we investigated the effect of particular pattern of rTMS on absent epileptic model .The epileptic model was induced by an acute injection of pentylenetetrazol in rats. The inhibitory effects of rTMS were investigated by evaluating 4 convulsive index including the latency until onset of stage 2 and 4, duration of generalized seizures and maximum seizure stage. Result indicated that application of particular pattern of rTMS 3 minute before injection of PTZ significantly inhibited PTZ-induced convulsions.

Vitamin D protects against hippocampal apoptosis related with seizures induced by kainic acid and pentylenetetrazol in rats

ObjectivesThe hippocampus is susceptible to damage in patients with epilepsy and in animals with seizures caused by excitotoxic agents. The effect of vitamin D on hippocampal apoptosis related with seizures has not been reported. However, epileptic patients have an increased risk of hypovitaminosis D which is most likely due to the effects of antiepileptic drugs. Therefore, in this study, it was aimed to evaluate the effects of vitamin D on hippocampal apoptosis related with seizures by using pentylenetetrazol (PTZ) and kainic acid (KA) in rats. MethodsMale Sprague Dawley rats, aged 5.5 weeks, were randomly divided into six groups: control, vitamin D, PTZ, KA, PTZ + vitamin D and KA + vitamin D groups. The groups that received vitamin D were given 500 IU/kg of vitamin D daily for two weeks in addition to a standard diet. At the end of this period, PTZ and KA were applied to trigger seizures in the rats in the seizure groups. 24 h after the administration of PTZ and KA, the rats were decapitated. In the hippocampal region, apoptosis was assessed by TUNEL and brain-derived neurotrophic factor (BDNF), Bax, caspase-3 and c-fos activation were evaluated by immunohistochemical method. ResultsBDNF level increased while c-fos, Bax and caspase-3 levels decreased (p < 0.0001, in all) in the hippocampal neurons of the groups that were pre-treated with vitamin D before the administration of PTZ and KA, in comparison with the PTZ and KA groups. Vitamin D significantly decreased the number of apoptotic cells in these rats in comparison with the PTZ and KA groups (p < 0.0001). ConclusionThis study indicates that vitamin D has neuroprotective effects on hippocampal apoptosis induced by PTZ and KA in rats. With this study it is suggested that keeping vitamin D levels within normal limits may be beneficial for patients with epilepsy, especially children.

Justicia spicigera Schltdl. and kaempferitrin as potential anticonvulsant natural products

Justicia spicigera Schltdl. is a vegetal species traditionally used to control epilepsy, but scientific evidence is required to reinforce this activity. The aim of the study was to evaluate the anticonvulsant-like activity of J. spicigera aqueous extract (JsAE) and a bioactive compound. JsAE was assessed in a dose-response manner (30, 100 and 1000mg/kg, i.p.) using the pentylenetetrazol (PTZ)-induced seizures and maximal electroshock seizure (MES) test in mice in comparison to ethosuximide (ETX, reference drug 100mg/kg, i.p.) or phenytoin (25mg/kg, i.p.), respectively. Then a significant dosage (1000mg/kg, i.p.) was chosen to examine electrographic activity (EEG) in rats. Treatment groups were compared to the vehicle and ETX in the convulsive behavior alone or simultaneous to EEG after PTZ-induced seizures (80 or 35mg/kg, i.p., mice or rats). Kaempferitrin (a flavonoid of JsAE) and ETX were administered via intracerebroventricular (i.c.v, 4th ventricle, 1μg/μL) and tested in the presence of PTZ in rats. Results confirmed that JsAE delayed the onset of seizures and reduced frequency of tonic convulsion and mortality in mice. JsAE or kaempferitrin also decreased the EEG spikes frequency and amplitude in a similar manner than EXT in rats. In conclusion, these preliminary data give evidence of the potential of J. spicigera as possible anticonvulsant as recommended in folk medicine for treating epilepsy, where kaempferitrin is suggested as a partial responsible bioactive compound.

Bumetanide reduce the seizure susceptibility induced by pentylenetetrazol via inhibition of aberrant hippocampal neurogenesis in neonatal rats after hypoxia-ischemia.

Hypoxia-ischemia brain damage (HIBD) is one of prevalent causes of neonatal mortality and morbidity. Our data demonstrated that hypoxia-ischemia (HI) induced Na+-K+-Cl--co-transporter 1 (NKCC1) increasing in hippocampus. Previous studies demonstrated that NKCC1 regulates various stages of neurogenesis. In this study, we studied the role of increased NKCC1 in regulating of HI-induced neurogenesis. HIBD model was established in 7days old Sprague-Dawley rat pup, and the expression of NKCC1 was detected by western blot and qPCR. Brain electrical activity in freely rats was monitored by electroencephalography (EEG) recordings. HI-induced neurogenesis was detected by immunofluorescence staining. Neurobehavioral test was to investigate the neuro-protective role of bumetanide, an inhibitor of NKCC1, on neonatal rats after HI. The results showed that bumetanide treatment significantly reduced brain electrical activity and the seizure stage of epilepsy induced by pentylenetetrazol (PTZ) in vivo after HI. In addition, bumetanide restored aberrant hippocampal neurogenesis and associated cognitive function. Our data demonstrated that bumetanide reduces the susceptibility of epilepsy induced by PTZ in rats suffering from HI injury during neonatal period via restoring the ectopic newborn neurons in dentate gyrus (DG) and cognitive function.

The Influence of Vagus Nerve and Spinal Cord Stimulation on the Ictal Fast Ripple Activity in a Spike-and-Wave Rat Model of Seizures.

Fast ripple (FR) activity has received increasing attention as a potential epileptic marker. The current knowledge on how neurostimulation affects FRs is, however, very limited. In this study, we assess the influence of the vagus nerve stimulation (VNS) and spinal cord stimulation (SCS) frequency on ictal FRs associated with spike-and-wave (SW) seizures. SW discharges were induced and maintained by an infusion of pentylenetetrazol in rat. During ongoing SW seizures, SCS was conducted at 30, 80, 130, and 180 Hz and VNS at 10, 30, 80, 130, and 180 Hz. The FRs were derived from intracortical recordings and the FR rate was used for quantifying the level of FR activity. The FR rate was significantly correlated (r = 0.81) with the level of total pentylenetetrazol dose. Compared with no stimulation intervals, SCS conducted at 80, 130, and 180 Hz significantly reduced the normalized FR rate by 24, 38, and 44%, respectively. Similarly, VNS conducted at 30, 80, 130, and 180 Hz significantly reduced the normalized FR rate by 23, 40, 61, and 65%, respectively. In the present model of sustained SW seizures, the FR rate was proportional with the severity of the SW seizures. Both SCS and VNS attenuated the FR rate and this attenuation was consistently strongest at the higher stimulation frequencies. Our results suggest that SCS may induce some of the same antiepileptic effects as VNS.

Acute seizure suppression by transcranial direct current stimulation in rats.

ObjectiveCathodal transcranial direct current stimulation (tDCS) is a focal neuromodulation technique that suppresses cortical excitability by low-amplitude constant electrical current, and may have an antiepileptic effect. Yet, tDCS has not been tested in status epilepticus (SE). Furthermore, a combined tDCS and pharmacotherapy antiseizure approach is unexplored. We therefore examined in the rat pentylenetetrazol (PTZ) SE model whether cathodal tDCS (1) suppresses seizures, (2) augments lorazepam (LZP) efficacy, and (3) enhances GABAergic cortical inhibition.MethodsExperiment 1 aimed to identify an effective cathodal tDCS intensity. Rats received intraperitoneal PTZ followed by tDCS (sham, cathodal 1 mA, or cathodal 0.1 mA; for 20 min), and then a second PTZ challenge. In Experiment 2, two additional animal groups received a subtherapeutic LZP dose after PTZ, and then verum or sham tDCS. Clinical and electroencephalography (EEG) epileptic activity were compared between all groups. In Experiment 3, we measured GABA-mediated paired-pulse inhibition of the motor evoked potential by paired-pulse transcranial magnetic stimulation (ppTMS) in rats that received PTZ or saline, and either verum or sham tDCS.ResultsCathodal 1 mA tDCS (1) reduced EEG spike bursts, and suppressed clinical seizures after the second PTZ challenge, (2) in combination with LZP was more effective in seizure suppression and improved the clinical seizure outcomes compared to either tDCS or LZP alone, and (3) prevented the loss of ppTMS motor cortex inhibition that accompanied PTZ injection.InterpretationThese results suggest that cathodal 1 mA tDCS alone and in combination with LZP can suppress seizures by augmenting GABAergic cortical inhibition.

Analysis of the Expressions of TrkB and GLT-1 in Brain Tissues on a Rat Pentylenetetrazol Kindling Model of Chronic Epilepsy.

Tyrosine kinase receptor-B (TrkB), a high-affinity receptor for brain-derived neurotrophic factor (BDNF), functions via specific binding with the BDNF. Although studies have shown that BDNF promotes the expression of glutamate transporter-1 (GLT-1) in the glial cells during neurodegeneration, evidence is lacking regarding whether BDNF/TrkB pathway is associated with the abnormal function of GLT-1.The aim of this study is to analyze the expressions of TrkB and GLT-1 in the brain of pentylenetetrazol (PTZ) kindling model of chronic epilepsy in rats, and to provide evidence for the correlation between the BDNF/TrkB pathway and GLT-1 and clues for the prevention and treatment targets of epilepsy. In total, 40 Sprague-Dawley rats were randomly classified as model (n= 30) and control (n= 10) groups respectively. Western blotting was used for analyzing the expression of TrkB and GLT-1, and double immunofluorescence staining was used for detecting the cells positive for both glial fibrillary acidic protein (GFAP) and TrkB in the hippocampus and temporal cortex 7 days after establishing the kindling model. Compared with the control group, protein level expression of TrkB and GLT-1, and mean optical density (OD) value of the cells positive for both GFAP and TrkB in the hippocampus and temporal cortex were significantly increased after the onset of epilepsy (P < 0.05). The BDNF/TrkB pathway may participate in the epileptogenesis through modulating the biological effect of GLT-1 in the glial cells of PTZ kindling rats, though the specific mechanisms require further investigations.

High dosage of agmatine alleviates pentylenetetrazole-induced chronic seizures in rats possibly by exerting an anticonvulsive effect.

The aim of the present study was to investigate the mechanism underlying the effects of different doses of agmatine in rats with chronic epilepsy. To generate chronic epilepsy models, rats pretreated with different doses of agmatine (20, 40 and 80 mg/kg) were intraperitoneally injected with pentylenetetrazole (35 mg/kg) for 28 consecutive days. Epileptic behavior was observed using behavioral measurements of convulsion for 1 h after each treatment with pentylenetetrazole. Morphological alterations of the hippocampal neurons were also observed using hematoxylin and eosin staining. In addition, the expression levels of glial fibrillary acidic protein and inducible nitric oxide synthase (iNOS) in the hippocampus were detected by immunohistochemistry. Furthermore, reverse transcription polymerase chain reaction was performed to detect the mRNA expression of two subunits (NR1 and NR2B) of the N-methyl-D-aspartic acid (NMDA) receptor in the rat hippocampus. The results demonstrated that administration of agmatine (80 mg/kg) significantly decreased the daily average grade of epileptic seizures and also reduced neuronal loss and astrocyte hyperplasia in the hippocampal area. Furthermore, agmatine (80 mg/kg) affected the mRNA expression levels of the NR1 subunit of the NMDA receptor, however, agmatine had no effect on the expression of iNOS in the hippocampus. Higher doses of agmatine inhibited the effect of pentylenetetrazole in rats, reduced astrocytic hyperplasia and neuronal damage in the hippocampus caused by seizures and selectively reduced the expression of the NR1 subunit of NMDA. Our results suggest that agmatine has an anticonvulsive effect in chronic epilepsy.

EFFECT OF SOME ANTIDIABETIC DRUGS ON BIOCHEMICAL PARAMETERS IN EXPERIMENTALLY INDUCED EPILEPTIC RATS

The present study was conducted to evaluate the effect of glibenclamide and metformin on some biochemical parameters in rats induced epilepsy. Male Wister was induced epilepsy by injection of pentylenetetrazole (PTZ) at a dose 100 mg/kg of body weight, the rats randomly divided into 3 groups (10-12 rat/group). The group 1: leaved without treatment and served as control group; Group 2: was treated with glibenclamide 5 mg/kg.b.w.; Group 3: was treated with metformin 150 mg/kg.b.w. All treatments were once daily for 1 week, blood samples were collected at 3, 24 hours, and week after induced of epilepsy. The results show that treated with PTZ leads to significant decrease in glucose level in all times after treatment, and significantly decreased level of cholesterol after 3 hours, and a week after treatment, while level of albumin was significantly decreased after a week of treatment, also PTZ treatment increased level of aspartate aminotransferase (AST) in all times after treatment, while level of alanine aminotransferase (ALT) was significantly increased after 3 hours, then significantly decreased after a week of treatment. PTZ treatment doesn't show any effect on levels of total protein, and globulin. Treatment with glibenclamide leads to significant increase level of glucose in all times after treatment, also level of cholesterol was significantly increased at 3, 24 hours after treatment with glibenclamide. Level of (AST) was significantly decreased in all times after treatment with glibenclamide, but level of (ALT) was increased only after a week of treatment, glibenclamide don’t cause any effect on levels total protein, albumin, and globulin. Treatment with metformin leads to significantly decreased level of glucose after 3, 24 hours of treatment, with significant increase after a week of treatment, while level of cholesterol was significantly increased after 3 hours, and significantly decreased after 24 hours of treatment. Levels of total protein and globulin were increased significantly after 3 hours only, level of albumin significant decrease after 24 hours treatment, also metformin lead to significantly decreased level of (AST) after 3 hours, and significantly increased after a week of treatment, while (ALT) level significantly increased after 24 hour, and week of treatment. These results indicate that glibenclamide and metformin have good roles in control of epilepsy-induced by PTZ in rats through several significant changes of biochemical parameters.

Cathodal transcranial direct current stimulation (tDCS) suppresses seizures, augments lorazepam potency, and enhances cortical inhibition in the rat pentylenetetrazol epilepsy model

Cathodal tDCS has antiepileptic potential. Yet whether its anti-seizure effect is via GABAergic mechanisms is unknown. We therefore tested in the pentylenetetrazol (PTZ) rat epilepsy model whether cathodal tDCS (1) suppresses seizures, (2) augments benzodiazepine efficacy, and (3) enhances GABAergic cortical inhibition.

Positive effect of calcitonin on the seizures induced by pentylenetetrazole in rats

There are many difficulties involved with the treatment of epilepsy, and these problems have driven the search for new agents to control epileptic seizures. Calcitonin is a peptide hormone that has been well studied and shown to have a positive effect on neuropathic and chronic pain. The mechanism by which calcitonin affects these pain syndromes is thought to be similar to the effect of antiepileptic drugs, such as pregabalin, gabapentin and carbamazepine. In this study, we aim to investigate the effects of calcitonin on seizures induced by pentylenetetrazole (PTZ) in rats. The rats were divided into four groups. The first group was the control group, and the rats were given no medications. The second group was given saline+PTZ. The third group was given 50IU/kg calcitonin+PTZ, and the fourth group was given 100IU/kg calcitonin+PTZ. EEG traces, Racine's convulsion stages and the time of onset of the first myoclonic jerk were compared between the groups. Between the groups, there were significant differences in the Racine's convulsion stages, the onset of the 'first myoclonic jerk', and the rate of the spikes in the EEG traces. The differences were more pronounced in the 100IU/kg calcitonin-treated group (p<0.001). It has been stated that calcitonin relieves pain via regulating voltage-gated Ca(2+) and/or Na(+) channels. Calcitonin has a positive effect on convulsions in epileptic rats, possibly using the same mechanisms as is used in the treatment of neuropathic and chronic pain.

P-Glycoprotein Contributes to Cell Membrane Depolarization of Hippocampus and Neocortex in a Model of Repetitive Seizures Induced by Pentylenetetrazole in Rats

P-glycoprotein (P-gp) has been associated with pharmacoresistance and mechanisms regulating the membrane potential. However, at present it is unknown if P-gp overexpression in brain is associated with changes in membrane depolarization in refractory epilepsy. Experiments were designed to evaluate the membrane depolarization and P-gp overexpression induced by repetitive pentilenetetrazole (PTZ)-induced-seizures. Wistar rats were daily treated with PTZ during 4 to 7 days (PTZ4 and PTZ7 groups), and the brain was used to evaluate membrane potential by in vitro electrophysiological procedures and using bis-oxonol dye, [bis-(1,3-dibutylbarbituric acid) trimethine oxonol (DiBAC4(3)], a fluorescence dye voltage-sensitive to membrane potentials. Rats with repetitive PTZ-induced seizures demonstrated lower phenytoin-induced anticonvulsant effects, increased number of DiBAC4(3) fluorescence cells and P-gp overexpression in hippocampus and neocortex, as well as augmentation of the induced fEPSP in CA1 field. These changes were more evident in PTZ7 group. Phenytoin or phenytoin plus nimodipine (a P-gp antagonist) avoided the enhanced fEPSP and decreased DiBAC4(3) fluorescence in animals from PTZ4 group. However, in PTZ7 group these effects were evident only when phenytoin was combined with nimodipine. An additional flow cytometry study demonstrated increased intracellular accumulation of DiBAC4(3) in K562 leukemic cells that overexpress MDR-1 and COX-2 genes, and are refractory to specific cytotoxic agents. These results represent the first evidence supporting the notion that brain P-gp overexpression contributes to a progressive seizure-related membranes depolarization in hippocampus and neocortex. Further experiments should be carried out to confirm the role of P-gp on membrane depolarization and epileptogenesis process.

Tamoxifen Mimics the Effects of Endogenous Ovarian Hormones on Repeated Seizures Induced by Pentylenetetrazole in Rats

In the present study, the effects of tamoxifen on pentylenetetrazole (PTZ)-induced repeated seizures and hippocampal neuronal damage in ovariectomized rats were investigated. Thirty seven virgin female Wistar rats were divided to: (1) control, (2) sham-PTZ, (3) sham-PTZ-tamoxifen (sham-PTZ-T), (4) Ovariectomized -PTZ (OVX-PTZ) and (5) OVX-PTZ-tamoxifen (OVX-PTZ-T) groups. The animals of groups 3 and 5 were injected by tamoxifen (10 mg/kg) on 7 consecutive days. After 7 days of tamoxifen injection, they also were then injected by tamoxifen 30 min prior each PTZ injection. PTZ (40 mg/kg) was injected on 6 consecutive days and the animal behaviors were observed for 60 min. The histological methods were then used to determine dark neurons in hippocampus. A significant decrease in the seizure score was seen in OVX-PTZ group compared to Sham-PTZ. The animals of OVX-PTZ-T group had a significant higher seizure score compared to OVX-PTZ group. The dark neurons in DG of OVX group were lower than sham group (p<0.01). The numbers of dark neurons in CA1 area of OVX-PTZ-T group was higher than OVX-PTZ group (p<0.05) compared to control, the numbers of dark neurons in CA3 area showed a significant increase in Sham-PTZ and OVX-PTZ group (p<0.05 and p<0.01 respectively). Dark neurons in OVX-PTZ-T group were higher than OVX-PTZ group (p<0.05). It is concluded that pretreatment of the ovariectomized rats by tamoxifen increased PTZ-induced seizure score and dark neurons. It might be suggested that tamoxifen has agonistic effects for estrogen receptors to change the seizure severity.

Contribution of estradiol in sex-dependent differences of pentylenetetrazole-induced seizures in rats

In the present study the contribution of estradiol in sex-dependent differences of pentylenetetrazole (PTZ)-induced seizures was investigated in rats. The rats were divided into four groups: 1) sham, 2) ovariectomized (OVX), 3) ovariectomized-estradiol (OVX-Est) and 4) male. The OVX-Est group received estradiol valerate (2 mg/kg; i.m/4 weeks) while, male, sham and OVX groups received vehicle. The animals were injected by PTZ (90 mg/kg). The latencies to minimal clonic seizures (MCS) and generalized tonic-clonic seizures (GTCS), were recorded. Serum 17β-estradiol and testosterone levels were also determined using an Elisa kit. GTCS latency in OVX rats was higher than in sham-operated animals (P < 0.05). MCS and GTCS latency in the male group was significantly higher than in the sham, OVX and OVX-Est groups (P < 0.001 and P < 0.01). There was no significant difference in MCS or GTCS latencies among OVX-Est, sham and OVX groups. Serum 17β-estradiol level in the OVX group was significantly lower than in the sham (P < 0.01) and in the OVX-Est group it was higher than in the sham, OVX and male groups (P < 0.01 and P < 0.001). Serum testosterone level in the male group was significantly higher than in all the other three groups (P < 0.001). It seems that testosterone probably has a more efficient role than estradiol in the gender dependent difference in seizure caused by PTZ in rats.

Anticonvulsant effect of Marsilea quadrifolia Linn. on pentylenetetrazole induced seizure: A behavioral and EEG study in rats

Ethnopharmacological relevanceMarsilea quadrifolia Linn (MQ) extract has been used traditionally as sedative and antiepileptic drug in India. Aim of this studyTo investigate the anticonvulsive potential of MQ extracts by using behavior and electroencephalographic (EEG) analysis on pentylenetetrazole (PTZ) induced seizure model in rats. Materials and methodsFor anticonvulsant effect, 60minutes after administration of MQ, behavior and EEG were analyzed during PTZ (60mg/kg) induced seizures. Changes of EEG power, latency of onset of seizure, seizure severity score, and duration of epileptic seizure were determined. ResultsBoth the water and ethanol extract of MQ increased the latency of seizure but also decreased duration of epileptic seizure and seizure severity score. This reduction of seizure severity was also observed in EEG recording and EEG power analysis. The effectiveness of MQ ethanol extract is better than MQ water extract. ConclusionBoth water and ethanol extract of MQ were effective in reducing the severity of behavioral and EEG seizures induced by PTZ in rats. This study justifies the traditional use of this plant in epilepsy.