Characterization of chemical ingredients and anticonvulsant activity of American skullcap (Scutellaria lateriflora).

American skullcap (the aerial part of Scutellaria lateriflora L.) has been traditionally used by Native Americans and Europeans as a nerve tonic, sedative, and anticonvulsant. However, despite some previous studies, the quality and safety, the bioactive ingredients, and the pharmacological properties of American skullcap are not fully understood. The aims of this study were to characterize the chemical ingredients of American skullcap and to evaluate its anticonvulsant activity. Twelve phenolic compounds including 10 flavonoids and two phenylethanoid glycosides were isolated and identified from American skullcap and used as marker compounds. An HPLC analytic method for analyzing these marker compounds in commercial American skullcap products from different sources was established and validated. The anticonvulsant activity of American skullcap was determined in rat models of acute seizures induced by pilocarpine and pentylenetetrazol. The results from this study indicate that (1) phenolic compounds, especially flavonoids, are the predominant constituents in American skullcap; (2) American skullcap products have similar constituents, but the content and relative proportions of the individual constituents varies widely; and (3) American skullcap has anticonvulsant activity in rodent models of acute seizures.

Possible mechanisms for the anticonvulsant activity of fructose-1,6-diphosphate.

Fructose-1,6-diphosphate (FDP), an intracellular metabolite of glucose, has anticonvulsant activity in several models of acute seizures in laboratory animals. The anticonvulsant effect of FDP is most likely due to a direct effect since intraperitoneal and oral administration results in significant increases in brain levels. A number of mechanisms have been proposed for this action of FDP. One possibility is that peripheral administration of FDP results in changes in brain metabolism that are anticonvulsant. Glucose can be metabolized through the glycolytic or pentose phosphate pathway. There is evidence that the pentose phosphate pathway is more active in the brain than in other tissues, and that, in the presence of elevated levels of FDP, the majority of glucose is metabolized by the pentose phosphate pathway. The pentose phosphate pathway generates NADPH, which is used to reduce glutathione. The reduced form of endogenous glutathione has been shown to have anticonvulsant activity. Taken together, the data suggest a hypothesis that exogenously administered FDP gets into the brain and astrocytes where it increases the flux of glucose through the pentose phosphate pathway, generating additional NADPH for the reduction of glutathione.

Oral administration of fructose-1,6-diphosphate has anticonvulsant activity.

Recently it has been shown that fructose-1,6-diphosphate (FDP) has dose-dependent anticonvulsant activity in rat models of acute generalized motor seizures induced with chemical convulsants. The present study asked whether FDP also has activity in an epileptic brain after oral administration and activity against non-convulsive seizures. Animals (n = 14) were administered pilocarpine to induce status epilepticus. Several weeks later, these animals had spontaneous seizures and a baseline rate of seizure frequency was determined over a 6-day period. Animals were then continued without treatment (n = 8) or 0.5% FDP was added to the drinking water (n = 6). In animals treated with FDP the seizures completely stopped after 7 days. Removal of FDP from the water resulted in the return of seizure activity in 4 of the 6 animals by 16 days of observation. To induce non-convulsive seizures, animals (n = 6) received a single injection of gamma-butyrolactone (GBL, 100 mg/kg i.p.). All animals had spike-wave activity recorded in the cortex within minutes after GBL administration. Administration of a single injection of FDP (500 g/kg i.p.) had no effect on the baseline cortical activity, nor on the spike-wave activity induced by GBL (n = 5). These experiments suggest that oral administration of FDP may have utility in the treatment of partial or generalized convulsive seizure disorders, but not absence seizures.

Antioxidants and free radical scavengers do not consistently delay seizure onset in animal models of acute seizures.

A number of herbal compounds with direct antioxidant activity slow the onset, or completely block, the occurrence of seizures. This increase in latency has been proposed to be due to the antioxidant activity. This hypothesis was directly tested by determining the effects of Trolox, a vitamin E analog, vitamin C, melatonin, and alpha-lipoic acid on the latency to acute seizures induced with pilocarpine, kainic acid, or subcutaneous pentylenetetrazol (PTZ) in adult rats. Trolox, vitamin C, and alpha-lipoic acid had significant anticonvulsant activity against pilocarpine, but there were no acute changes in reduced glutathione levels at 15 or 120 minutes. Other than reduced mortality with vitamin C in the PTZ model, none of the antioxidants had a significant effect against PTZ- or kainic acid-induced seizures. The lack of consistent anticonvulsant effect suggests that the antioxidant activity of the herbal preparations cannot account for the delay in seizure onset.

Association of NRH:quinone oxidoreductase 2 gene promoter polymorphism with higher gene expression and increased susceptibility to Parkinson's disease.

The N-ribosyldihydronicotinamide (NRH):quinone oxidoreductase 2 (NQO2) gene encodes an enzyme that catalyzes activation of quinones. Blood DNA from 80 control individuals and 118 age-matched Parkinson's disease patients were analyzed for NQO2 gene promoter polymorphisms. The results revealed three allelic variants, designated I-29, I-16, and D. These results were confirmed in fibroblast cell lines. In patients with Parkinson's disease, there was a significant increase in the frequency of the D allele, but there was no difference in the frequency of the alleles in familial compared to sporadic Parkinson's disease. The D and I-16 promoters direct higher NQO2 gene expression that results in higher enzyme activity. Overexpression of NQO2 in the catecholaminergic neuroblastoma SH-SY5Y cells resulted in increased production of reactive oxygen species when exposed to exogenous dopamine. The results suggest that the association of the D promoter with Parkinson's disease may be due to an increase in expression of the NQO2 gene.

Pharmacokinetics of fructose-1,6-diphosphate after intraperitoneal and oral administration to adult rats.

Exogenously administered fructose-1,6-diphosphate (FDP) has been studied for its ability to protect tissue during hypoxia or ischemia. Recently, a clear effect of FDP on the central nervous system has raised the question whether FDP can get into the brain. FDP levels were measured in blood, brain, liver, kidney, muscle and fat after intraperitoneal administration of a single 0.5gkg(-1) dose of FDP to adult male Sprague-Dawley rats. A complete time course of the levels in blood and brain was determined. The levels of FDP in the blood and brain increase simultaneously, i.e. there is no lag in the increase in the brain. The levels of FDP fall to baseline in liver, kidney, muscle and fat by 12h, but remain elevated in blood and brain. However, levels in the blood at 12h are significantly decreased from the peak levels, while those in brain are not different from the peak levels, suggesting that the kinetics of FDP in blood and brain are quite different. Stripping the endothelial cells from the brain tissue sample did not change the levels of FDP indicating that FDP is not trapped in the capillary cells. Incubation of brain slices in a solution of FDP, followed by washing, raised tissue levels of FDP indicating that FDP is taken up into cells within the brain. Finally, the experiments demonstrate a significant increase in brain levels of FDP after oral administration. These data suggest that an oral formulation of FDP might be developed for treatment of neurological disease.

Fructose-1,6-bisphosphate has anticonvulsant activity in models of acute seizures in adult rats.

A variety of observations suggest that decreasing glycolysis and increasing levels of reduced glutathione, generated by metabolism of glucose through the pentose phosphate pathway, would have an anticonvulsant effect. Because fructose-1,6-bisphosphate (F1,6BP) shifts the metabolism of glucose from glycolysis to the pentose phosphate pathway, it was hypothesized to have anticonvulsant activity. The anticonvulsant activity of F1,6BP was determined in rat models of acute seizures induced by pilocarpine, kainic acid, or pentylenetetrazole. The efficacy of F1,6BP was compared with that of 2-deoxyglucose (2-DG; an inhibitor of glucose uptake and glycolysis), valproic acid (VPA), and the ketogenic diet. One hour before each convulsant, Sprague Dawley rats received either saline (as seizure controls), F1,6BP (0.25, 0.5 or 1 g/kg), 2-DG (0.25 or 0.5 g/kg), or VPA (0.3 g/kg). Additional animals received the ketogenic diet (starting at 20 or 60 d old). Time to seizure onset, seizure duration, and seizure score were measured in each group. F1,6BP had dose-dependent anticonvulsant activity in all three models, whereas VPA had partial efficacy. 2-DG was only effective in the pilocarpine model. The ketogenic diet had no effect in these models. F1,6BP was also partially effective when given at the first behavioral seizure after pilocarpine. Administration of sodium lactate, which bypasses the block in the glycolytic pathway, abolished the anticonvulsant activity of 2-DG in the pilocarpine model, but only decreased the efficacy of F1,6BP. These data demonstrate that F1,6BP has significant anticonvulsant efficacy.

Regulation of extracellular calcium in the hippocampus in vivo during epileptiform activity--role of astrocytes.

Astrocytes have been suggested to regulate the extracellular calcium concentration ([Ca(2+)](o)), but this has not been thoroughly investigated. Adult, male Sprague-Dawley rats were used to record changes in [Ca(2+)](o) in the hippocampus during epileptiform activity. Maximal decreases in [Ca(2+)](o) in CA1 were measured in the pyramidal cell layer during 20 Hz, 20s stimulus trains to the contralateral CA3 region. Maximal decreases in [Ca(2+)](o) in the dentate gyrus were measured when maximal dentate activation had appeared-irrespective of the location, frequency or duration of the stimulation. Maximal decreases were 36% greater in the dentate gyrus than in CA1. During prolonged discharges, [Ca(2+)](o) recovered partially towards the baseline in both hippocampal regions. To investigate the role of astrocytes, local injections of fluorocitrate (FC), a metabolic toxin selectively taken up by astrocytes, were used. FC (0.1, 0.25 or 0.5mM FC), but not vehicle (2 microl), caused a small, but significant decrease in the maximal changes in CA1, but an increase in the dentate gyrus. The results suggest that maximal decreases in [Ca(2+)](o) occur in the hippocampus in response to burst firing of neurons and that astrocytes play a minimal role in the regulation of [Ca(2+)](o) during epileptiform activity.

Anticonvulsant and neuroprotective effects of ginsenosides in rats.

A partially purified extract from American ginseng has been shown to have anticonvulsant activity. To identify the active components in this extract, the activities of the individual ginsenosides (Rb(1), Rb(3) and Rd), mixtures of the purified ginsenosides and a newly prepared Rb fraction were determined. One hour after treatment with vehicle or one of the ginseng products, seizures were induced in adult, Sprague-Dawley rats with kainic acid (KA, 10 mg/kg), pilocarpine (300 mg/kg) or pentylenetetrazole (PTZ, 50mg/kg i.p. or 90 mg/kg s.c.). Time to seizure onset, duration of seizure activity and seizure severity were determined. Weight change and neuronal damage were assessed 24h after administration of KA or pilocarpine. Mixtures of purified Rb(1), Rb(3) with or without Rd had significant anticonvulsant effects in all three models of acutely induced seizures demonstrating that the ginsenosides are the active components in the Rb extract. The individual ginsenosides significantly increased the latency to onset of seizures after administration of kainic acid. Since no one individual ginsenoside accounted for the majority of the activity of the Rb extract, the results suggest that the most effective anticonvulsant product is a combination of ginsenosides. In addition, all of the ginseng products had significant neuroprotective activity beyond the reduction in seizure severity and duration.

Protective effects of ginseng components in a rodent model of neurodegeneration.

To test the proposed neuroprotective activity of whole ginseng extract and its components, we used 3-nitropropionic acid (3-NP), an inhibitor of succinate dehydrogenase, to produce neurodegeneration. Treatment with 3-nitropropionic acid (90 mg/kg) over a 5-day period resulted in severe impairment of movement and loss of neurons in the striatum. Pretreatment with a preparation from the whole root of American ginseng had no protective effects. Pretreatment with a preparation of ground leaves and stems, which contains greater levels of ginsenosides than ground root, improved the behavioral score and reduced the volume of the striatal lesion. A partial purification of American ginseng was performed to concentrate the putative protective components: Rb1, Rb3, and Rd (termed Rb extract). Pretreatment with the Rb extract significantly reduced the 3-nitropropionic acid-induced motor impairment and cell loss in the striatum, and it completely prevented any mortality. Significant effects on motor function, mortality, and the striatal lesion volume also were measured in animals pretreated with the individual ginsenosides, Rb1, Rb3, or Rd. The results demonstrate that some of the ginsenosides have neuroprotective activity, and that a partial purification of whole ginseng to concentrate the neuroprotective components may have utility as a neuroprotective agent.

Anticonvulsant activity of ginseng on seizures induced by chemical convulsants.

PURPOSE: To test the anticonvulsant activity of three preparations of American ginseng: whole root extract, whole leaves/stems extract, and a partially purified extract that concentrates the Rb ginsenosides (Rb extract). METHODS: One hour after treatment with normal saline, or one of the three ginseng preparations, seizures were induced in adult, male, Sprague-Dawley rats with kainic acid (KA; 10 mg/kg), pilocarpine (300 mg/kg, preceded by methylscopolamine, 1 mg/kg, s.c.), or pentylenetetrazol (PTZ, 50 mg/kg). Time to onset of seizure activity, duration of seizure activity for PTZ, seizure severity, and weight change for KA and pilocarpine were determined for each animal. The brains from animals who had received KA or pilocarpine were examined for severe neuronal stress, by using immunoreactivity for heat-shock protein (HSP)72. RESULTS: The Rb extract had a dose-dependent anticonvulsant effect in all three models of chemically induced seizures: increasing the latency to the seizures; decreasing the seizure score, weight loss, and subsequent neuronal damage after pilocarpine; and shortening the seizure duration and reducing mortality after PTZ. The Rb extract also significantly reduced the effects of KA, including completely blocking behavioral seizures. The root preparation increased the mortality rate after administration of pilocarpine, but had no other significant effects. The leaves/stems preparation, at 120 mg/kg, reduced the weight loss after pilocarpine, but had no other significant effects. CONCLUSIONS: Ginseng extract made from either the root or leaves/stems is ineffective against chemically induced seizures. A partial purification of the whole extract that concentrates the Rb1 and Rb3 ginsenosides has significant anticonvulsant properties.

Inhibition of aconitase in astrocytes increases the sensitivity to chemical convulsants.

Although there is evidence that astrocytes support neuronal function, the contribution of astrocytes to seizure onset and termination is not known. To determine whether there are changes in seizure susceptibility or neuronal damage when the ability of astrocytes to generate ATP is reduced, 0.5 nmol of fluorocitrate (FC) was injected into the right ventricle. Injection of FC alone did not produce electrographic or behavioral seizures and did not stress or injure neurons or astrocytes, as measured with silver stain and immunohistochemistry for HSP32 or HSP72. However, in animals pretreated with FC, administration of kainic acid, at a dose that does not initiate seizures in control animals (7 mg/kg), caused wet dog shakes and neuronal damage in the hilus. Wet dog shakes did not cause any neuronal damage in control animals. If the dose of FC was increased to 0.75 nmol, then subsequent administration of the same dose of kainic acid (7 mg/kg) caused stage 3-5 seizures. Injection of FC also reduced the dose of pilocarpine needed to produce seizures. Given simultaneously with FC, isocitrate, which bypasses the biochemical inhibition of aconitase, blocked the effects of FC in both kainic acid and pilocarpine treated animals. The data demonstrate that inhibition of aconitase in astrocytes lowers the doses of both kainic acid and pilocarpine that will cause behavioral seizures and may increase neuronal vulnerability to seizures.

Energy failure in astrocytes increases the vulnerability of neurons to spreading depression.

A neuroprotective role of astrocytes has been hypothesized, but the mechanism is debated and in vivo evidence is limited. To test this hypothesis, a sublethal stressor (spreading depression) and fluorocitrate (FC), a selective inhibitor of the astrocytic Krebs cycle, were used in urethane-anaesthetized adult rats. Neuronal damage was assessed 24 h after treatment with silver stain and immunoreactivity for a 72-kDa heat-shock protein. ATP levels and mitochondrial aconitase activity, a marker indicating exposure to reactive oxygen species, were measured after 4 and 24 h. Spreading depression alone did not affect ATP levels, mitochondrial aconitase activity, or induce neuronal injury in the cortex. Local or intraventricular injection of FC significantly decreased ATP levels and mitochondrial aconitase activity, but did not produce neuronal damage. In animals receiving injections of FC and then spreading depression, there was evidence of significant neuronal stress and damage. Isocitrate, which bypasses the metabolic inhibition produced by FC, prevented all of the changes seen after the combination of FC and spreading depression. One-hour pretreatment with dimethyl sulfoxide (a scavenger of hydroxyl radicals), deferoxamine (an iron chelator) or fructose-1,6-bisphosphate also blocked inactivation of mitochondrial aconitase, ATP depletion and the neuronal damage induced by FC and spreading depression. These experiments demonstrate that inhibition of the metabolism of astrocytes, with a decrease in ATP levels, will increase the susceptibility of neurons to the stress induced by spreading depression. The neuroprotective effects of dimethyl sulfoxide, deferoxamine and fructose-1,6-bisphosphate suggest that oxidative stress contributes to the neurotoxicity in this situation.

Astrocytes contribute to regulation of extracellular calcium and potassium in the rat cerebral cortex during spreading depression.

This study used spreading depression (SD), which is characterized by redistribution of ions, to examine the role of astrocytes in the regulation of extracellular potassium ([K+]o) and calcium ([Ca2+]o) levels. Recurrent spreading depression episodes were induced by application of 3 M potassium chloride to the cortex of adult anesthetized rats while monitoring the extracellular direct current (DC) potential shifts and changes in [K+]o or [Ca2+]o 6-7 mm away. The reversible glial toxins, fluorocitrate (FC) and fluoroacetate (FA), were injected locally into the cortex at doses that are selective for reducing glial function. The peak changes and area under the curve for [K+]o and [Ca2+]o, recovery rate for [K+]o, and interval between spreading depression episodes were measured before and at various times after administration of the toxins. Both fluorocitrate and fluroacetate slowed the recovery of the [K+]o and altered the recovery of the [Ca2+]o. Local injection of glutamate uptake inhibitors or barium had no effect on the peak changes in [K+]o or the rate of recovery of the [K+]o. The slowing of the recovery rate is consistent with the hypothesis that glial cells play a role in the return of [K+]o to baseline after spreading depression in the cortex in vivo. The change in movement of calcium after administration of FC suggests that astrocytes normally extrude calcium during spreading depression, resulting in rapid recovery of the levels of [Ca2+]o with an overshoot. These findings demonstrate that astrocytes contribute to the regulation of both potassium and calcium during and after a stress to the ionic homeostatic mechanisms.

Presence and induction of the enzyme NAD(P)H: quinone oxidoreductase 1 in the central nervous system.

NAD(P)H:quinone oxidoreductase 1 (NQO1) catalyzes a reductive detoxification that is thought to protect cells against the adverse effects of quinones and related compounds. NQO1 activity is present in all tissues. Absence of the enzyme produces abnormalities in the redox state and seizures, suggesting an important role of the protein in the central nervous system. Immunohistochemical analysis showed that the protein was found throughout the brain of the adult rat and mouse, with complete absence of the protein in brains from NQO1-/- mice. NQO1 was not seen in any neuronal population, but was localized to Bergmann glial in the cerebellum and a subset of the oligodendrocytes throughout the brain. Prolonged seizures induced in adult rats with kainic acid resulted in an increase in activity of the enzyme throughout the brain, most prominently in the cerebellum, but immunoreactivity did not appear in neurons. Comparison of the axons in the corpus callosum from a wild-type mouse to a knockout mouse showed that myelin is produced in the absence of NQO1, but there appears to be more small-diameter axons in the knockout animal. These results suggest that NQO1 has a role in myelination in the central nervous system or in the insulating/wrapping function of glial cells.

Epileptogenic activity of granulomas associated with murine cysticercosis.

Neurocysticercosis, caused by Taenia solium, is a common cause of neurologic disease in developing countries and among immigrants to the United States. Seizures are the most common clinical manifestation of neurocysticercosis. Imaging studies of patients with seizures from neurocysticercosis typically reveal evidence of an inflammatory reaction associated with the parasite or calcified granulomas. This study investigated whether a substance produced by the host granulomatous reaction to the dying parasite, in a mouse model of the infection, is sufficient to induce epileptiform activity. Granulomas associated with Taenia crassiceps cysticerci were removed from the peritoneal cavity of infected mice. One piece of the granuloma was used for blinded histological staging of the dying parasite. The second piece was used to generate extracts, which were injected into the hippocampus of an anesthetized Sprague-Dawley rat. Positive controls included animals injected with kainic acid, picrotoxin, or bicuculline. Seizures were recorded after injection of extracts from 6 out of 6 early stage granulomas, but only 1 out of 9 late stage granulomas. Injections of buffered saline, extracts from non-stimulated mouse spleen cells, and homogenates of viable parasite material caused no epileptiform activity. The data suggest that a substance in the granulomas early in the inflammatory response to the dying parasite is capable of inducing seizure activity. Further experiments are needed to dissect out the exact seizure mediator in the granuloma extracts.

Effects of glial toxins on extracellular acidification in the hippocampal CA1 region in vivo.

In the pyramidal cell layer of the CA1 region of the hippocampus in the urethane-anesthetized adult rat, there is an initial alkalinization followed by an acidification in response to synchronized seizure activity induced by stimulus trains. In this study, the role of astrocytes in these extracellular pH changes during neuronal activity was examined using local injection of two relatively selective glial toxins (fluorocitrate (FC) and fluoroacetate (FA)) into the CA1 cell layer. Both glial toxins reduced the peak level of acidification reached after 20 Hz stimulus trains to the contralateral CA3 region, without changing the lengthening of the afterdischarge, when compared to animals that had received a local injection of vehicle. After administration of either glial toxin, the peak level of acidification still correlated with the total discharge duration, but the levels of acidification were consistently lower than in control animals. Administration of either glial toxin had no effect on the peak alkalinization during the stimulus train, or on the rate of recovery from peak level of acidification. Injection of either vehicle, FA, or FC had no effect on the amplitude or frequency of the neuronal discharge during the afterdischarge. The results suggest that, in normal conditions, astrocytes contribute to the acidification of the extracellular space that occurs in response to intense neuronal activity. This acidification may contribute to feedback regulation of neuronal excitability.

Modulation of the in vivo effects of gabapentin by vigabatrin and SKF89976A.

Gabapentin (GBP) has been shown to reduce paired-pulse inhibition in the dentate gyrus of the urethane-anesthetized rat, which is a proconvulsant effect, and to shorten the afterdischarge duration, which is an antiepileptic effect. The mechanism by which GBP exerts these effects is not known, but a number of possibilities have been proposed. Here we tested the ability of vigabatrin (VGB), a GABA transaminase inhibitor, and SKF89976A, a selective GAT-1 blocker, to alter the effectiveness of GBP in the dentate gyrus in urethane-anesthetized adult Sprague-Dawley rats. VGB, alone at 100 mg/kg, had no effect on the evoked potentials or paired-pulse inhibition in the dentate gyrus, but did block lengthening of the afterdischarge. Pretreatment with VGB had no effect on the ability of GBP to reduce paired-pulse inhibition, but blocked the effect of GBP on seizure duration. SKF89976A, alone at 10 mg/kg, increased paired-pulse inhibition and blocked the lengthening of the afterdischarge in the seizure model. Pretreatment with SKF89976A had no effect on the actions of GBP on either paired-pulse inhibition or seizure duration. These results suggest that the action of GBP is not mediated through an inhibition of the GAT-1 transporter and probably not through an increase in basal levels of GABA. The data also suggest that the combination of VGB and GBP may be clinically less effective than the use of GBP alone.

Developmental expression of serum response factor in the rat central nervous system.

Serum response factor (SRF), a transcription factor known to be essential for early embryonic development as well as post-natal regulation of both cellular proliferation and myogenic differentiation, is expressed broadly in neurons within the adult mammalian central nervous system (CNS). The function of SRF within the developing CNS is not well established, but it is likely to play an important role in neuraxial development and neuronal function, since many of its known target genes (e.g., c-fos) and transcriptional partners (e.g., Elk-1) are also highly expressed in neurons. Immunohistochemical survey of the post-natal developing rat brain revealed a progressive increase in SRF immunoreactivity in neurons of the cerebral and cerebellar cortices, and in selective subcortical regions from birth (P0) through post-natal day 28 (P28). SRF immunoreactivity stabilized from P28 into adulthood. A few loci, such as the nucleus of cranial nerve VII, showed the reverse expression pattern (strong immunoreactivity at P0-P7, declining by P28). The developmental expression pattern of SRF overlaps significantly with that of myotonic dystrophy protein kinase, a potential upstream regulator, and of the LIM-only genes Lmo1, Lmo2 and Lmo3, whose products belong to a family of proteins known to be strong positive regulators of SRF's transcriptional activity. These data suggest that SRF has a significant function in the early post-natal development of the CNS.

Effects of antiepileptic drugs on extracellular pH regulation in the hippocampal CA1 region in vivo.

Intracellular and extracellular pH are known to influence neuronal activity and may play a role in seizure termination. In the pyramidal cell layer of the CA1 region of the hippocampus in the urethane anesthetized adult rat, there is an initial alkalinization in response to stimulus trains administered to the contralateral CA3 region. This is followed by an acidification that peaks after termination of the afterdischarge. Initial experiments demonstrated that the peak level of acidification correlated with the duration of the afterdischarge, but that the peak level of alkalinization did not. The effects of several antiepileptic drugs on the initial alkalinization were determined. Systemic administration of acetazolamide (50 mg/kg, n=4) and topiramate (45 mg/kg, n=7) and local administration of benzolamide (n=3), all of which inhibit carbonic anhydrase, decreased the initial alkalinization that occurs during the stimulus train. Diazepam (3 mg/kg, n=5) and phenobarbital (60 mg/kg, n=6), agonists at the GABA(A) receptor complex, increased the initial alkalinization, while sodium channel blockers phenytoin (80 mg/kg, n=5) and carbamazepine (50 mg/kg, n=5) had no significant effect. The data suggest that the alkalinization in CA1 in vivo is predominantly regulated through activity of the GABA(A) receptor, rather than through activation of glutamatergic receptors. The change in alkalinization does not appear to be related to the mechanism of the antiepileptic effect of the drugs that were tested.