Anticonvulsant Effects Of Ethanol Research Articles

Sex differences in effects of mild chronic stress on seizure risk and GABA A receptors in rats

Social stress is a common occurrence in our society that can negatively impact health. Therefore, we wanted to study the effects of a mild stressor designed to model social stress on seizure susceptibility and GABA A receptors in male and female rats. The mild chronic stress of individual housing consistently decreased bicuculline (but not pentylenetetrazol, PTZ) seizure thresholds by 10–15% in both sexes. Housing conditions did not alter the anticonvulsant activity of diazepam or ethanol, although the anticonvulsant effect of ethanol was significantly greater against PTZ-induced seizures. Experiments testing the addition of an acute restraint stress unmasked sex differences in seizure induction. The acute stress also selectively decreased the potency of GABA to modulate GABA A receptor-mediated chloride uptake in group-housed females. There were additional sex differences by housing condition for GABA A receptor-gated chloride uptake but no differences in [ 3H]flunitrazepam binding. We also found significant effects of sex and housing on ethanol-induced increases in corticosterone (CORT) levels. In summary, there were complex and sex-selective effects of mild chronic stress on seizure induction and GABA A receptors. Gaining a better understanding of mechanisms underlying interactions between sex and stress has important implications for addressing health concerns about stress in men and women.

Direct Evidence for a Cause-Effect Link Between Ethanol Potentiation of GABAA Receptor Function and Intoxication From Hyperbaric Studies in C57, LS, and SS Mice

Background: This article uses a direct ethanol antagonist, increased atmospheric pressure, to further test the causative link between ethanol potentiation of γ-aminobutyric acid (GABA) type A receptor function and ethanol's behavioral effects. This was done by determining whether initial biochemical findings in long-sleep (LS) mice extended to other genotypes and whether the previously reported insensitivity of short-sleep (SS) mice to pressure antagonism of ethanol-induced loss of righting reflex extended to a nonselected ethanol-induced behavior. Methods: The effects of 12 times normal atmospheric pressure of helium-oxygen gas (heliox) versus ethanol (25–200 mM) potentiation of GABA-activated Cl− uptake in brain membranes (microsacs) from C57, LS, and SS mice were tested by using a 36Cl− flux assay. The effects of pressure versus ethanol's (2 g/kg) anticonvulsant effect in SS mice were tested by using time to onset of isoniazid-induced myoclonic seizures. Results: Exposure to 12 times normal atmospheric pressure heliox antagonized ethanol potentiation of GABA-activated Cl− uptake in all three genotypes across a range of ethanol concentrations that cause ethanol's behavioral and anesthetic effects. Pressure did not affect baseline receptor function. The threshold for initiating ethanol potentiation differed between genotypes in accordance with their behavioral sensitivities to ethanol (C57 and LS, ≤25 mM; SS, >50 mM). Pressure antagonized ethanol's anticonvulsant effect in SS mice. Conclusions: The results add important direct evidence supporting the hypothesis that ethanol potentiation of GABAA receptor function is an initial action of ethanol causing its behavioral effects. These findings also provide insight into possible effects of selective breeding on GABAA receptor function.

Direct Evidence for a Cause‐Effect Link Between Ethanol Potentiation of GABAA Receptor Function and Intoxication From Hyperbaric Studies in C57, LS, and SS Mice

This article uses a direct ethanol antagonist, increased atmospheric pressure, to further test the causative link between ethanol potentiation of gamma-aminobutyric acid (GABA) type A receptor function and ethanol's behavioral effects. This was done by determining whether initial biochemical findings in long-sleep (LS) mice extended to other genotypes and whether the previously reported insensitivity of short-sleep (SS) mice to pressure antagonism of ethanol-induced loss of righting reflex extended to a nonselected ethanol-induced behavior. The effects of 12 times normal atmospheric pressure of helium-oxygen gas (heliox) versus ethanol (25-200 mM) potentiation of GABA-activated Cl- uptake in brain membranes (microsacs) from C57, LS, and SS mice were tested by using a 36Cl- flux assay. The effects of pressure versus ethanol's (2 g/kg) anticonvulsant effect in SS mice were tested by using time to onset of isoniazid-induced myoclonic seizures. Exposure to 12 times normal atmospheric pressure heliox antagonized ethanol potentiation of GABA-activated Cl- uptake in all three genotypes across a range of ethanol concentrations that cause ethanol's behavioral and anesthetic effects. Pressure did not affect baseline receptor function. The threshold for initiating ethanol potentiation differed between genotypes in accordance with their behavioral sensitivities to ethanol (C57 and LS, < or =25 mM; SS, >50 mM). Pressure antagonized ethanol's anticonvulsant effect in SS mice. The results add important direct evidence supporting the hypothesis that ethanol potentiation of GABA(A) receptor function is an initial action of ethanol causing its behavioral effects. These findings also provide insight into possible effects of selective breeding on GABA(A) receptor function.

Influence of ethanol on the pentylenetetrazol-induced kindling in rats.

The effects of ethanol on the development of pentylenetetrazol (PTZ)-kindling as well as on fully PTZ-kindled convulsions in rats were investigated. Ethanol (0.5, 1.0 and 1.5 g/kg i.p.) administered 15 min prior to each PTZ-injection (35 mg/kg i.p.; 3 times/week) significantly inhibited the progressive seizure development compared to saline-treated controls. For the higher doses of ethanol the kindling process was restricted to seizure stages of 1 or 2. Tolerance to this antiepileptogenic action did not occur even after 20 PTZ-stimulations. In a second series of experiments, 0.5 g/kg ethanol administered 10h before each PTZ-injection facilitated the rate of kindling development after 7 to 10 PTZ-injections, while the higher doses of ethanol did not modulate or even slightly reduced the seizure development. In a third test, intermittent administration of a high dose of ethanol (2 g/kg p.o.; twice daily for 6 days) before the kindling procedure (0.5 g/kg i.p. ethanol 10h prior to each PTZ-injection), significantly intensified the kindling development. In addition, studies with fully PTZ-kindled rats demonstrated that ethanol (0.1 to 1.5 g/kg i.p.), given 15 min prior or 2 min after PTZ, reduced the seizure severity in a dose-dependent manner. In conclusion, the present findings provide evidence for pronounced antiepileptogenic and anticonvulsant effects of ethanol after acute application, whereas repeated administration of high doses with longer withdrawal periods leads to proconvulsant actions, possible mediated via neuroadaptive changes in NMDA and/or GABA(A) receptor-related mechanisms.

Reduced Tolerance to Certain Pharmacological Effects of Ethanol After Chronic Administration in Perinatally Undernourished Rats

We have previously reported that recovered adult rats undernourished at perinatal age failed to develop tolerance to the anticonflict effect of ethanol after chronic ethanol administration (1 g/kg/day during 30 days) [4]. To further study the extent of this finding, we examined the effect of a similar chronic ethanol treatment on the hypothermic and anticonvulsant effects of ethanol in perinatally deprived rats. Hypoalgesic activity was assessed in ethanol treated rats during 15 days. After chronic ethanol treatment, a similar development of tolerance to the hypothermic effect of ethanol was observed in control and deprived rats. However, tolerance to the anticonvulsant and hypoalgesic effect of ethanol was significantly reduced in deprived as compared with control animals. Thus, early undernutrition differentially affects the development of tolerance elicited by chronic ethanol administration.

Contingent drug tolerance: Differential tolerance to the anticonvulsant, hypothermic, and ataxic effects of ethanol

The kindled-convulsion model of epilepsy was used to study contingent tolerance to ethanol's (1.5 g/kg; IP) anticonvulsant, hypothermic, and ataxic effects in adult male rats. In the present experiments, three groups of amygdala-kindled rats received a series of bidaily (one every 48 h) convulsive stimulations: one group received ethanol 1 h before each stimulation; one group received ethanol l h after each stimulation; and another group served as the saline control. Tolerance to ethanol's anticonvulsant effect (Experiments 1 and 2) was greatest in those rats that received ethanol before each convulsive stimulation; whereas, tolerance to ethanol's hypothermic (Experiments 1 and 2) and ataxic (Experiments 2) effects developed in both groups that received ethanol. These results were predicted on the basis of the drug-effect theory of drug tolerance: the theory that functional drug tolerance is an adaptation to the disruptive effects of drugs on concurrent patterns of neural activity, not to drug exposure per se.

Prenatal ethanol exposure: susceptibility to convulsions and ethanol's anticonvulsant effect in amygdala-kindled rats.

The present experiments assessed the effects of prenatal ethanol exposure on the susceptibility to convulsions and on the anticonvulsant effect of ethanol using the electrical kindling model of epilepsy in rats. Adult male Sprague-Dawley rats from prenatal ethanol (E), pair-fed (PF), and ad libitum-fed control (C) treatment groups were tested following the implantation of a stimulation electrode in the left amygdala complex. The same rats were tested in four consecutive experiments. Both E and PF rats showed a slightly slower rate of kindling than C rats, as measured by convulsion class but not as measured by forelimb clonus duration (experiment 1). However, the groups did not differ significantly in the electrical stimulation threshold for kindled convulsions (experiment 2). Furthermore, prenatal ethanol exposure had no significant effect on the dose-response curve for ethanol's (0, 0.9, 1.1, 1.3, and 1.5 g/kg, ip) anticonvulsant effect (experiment 3), or on the rate of tolerance development to ethanol's (1.5 g/kg, ip) anticonvulsant effect (experiment 4) on kindled convulsions. Thus, prenatal exposure to ethanol does not appear to have long-term effects on the susceptibility to convulsions or on the anticonvulsant effect of ethanol in adult male rats in the kindling model as used in the present experiments.

Low‐Level Hyperbaric Antagonism of Ethanol's Anticonvulsant Property in C57BL/6J Mice

This study investigated the ability of hyperbaric exposure to antagonize ethanol's anticonvulsant effect on isoniazid (INH)-induced seizures. Drug-naive, male C57BL/6 mice were injected intraperitoneally with saline, 1.5, 2.0, or 2.5 g/kg ethanol followed immediately by an intramuscular injection of 300 mg/kg of INH. The mice were then exposed to either 1 atmosphere absolute (1 ATA) air, 1 ATA helium-oxygen gas mixture (heliox), or 12 ATA heliox at temperatures that offset the hypothermic effects of helium. Ethanol increased the latency to onset of myoclonus in a dose-dependent manner. Exposure to 12 ATA heliox antagonized ethanol's anticonvulsant effect at 2.0 and 2.5 g/kg, but not at 1.5 g/kg. Ethanol also increased the latency to onset of clonus in a dose-dependent manner beginning at 2.0 g/kg. Exposure to 12 ATA heliox antagonized this anticonvulsant effect. When exposed to 12 ATA heliox, the blood ethanol concentrations at time to onset of myoclonus were significantly higher in mice treated with 2.5 g/kg of ethanol as compared with blood ethanol concentrations of mice exposed to 1 ATA air. These findings extend the acute behavioral effects of ethanol known to be antagonized by hyperbaric exposure and support the hypothesis that low-level hyperbaric exposure blocks or reverses the initial action(s) of ethanol leading to its acute behavioral effects.

Investigation of the involvement of opioid receptors in the action of anticonvulsants.

This study investigates the possible involvement of opioid receptors in the action of a variety of anticonvulsant agents. The opioid antagonist naloxone (0.3, 1 mg/kg IP) and the selective mu-opioid antagonist cyprodime (3 mg/kg IP) significantly inhibited the increase in electroshock seizure threshold induced by phenytoin (3 mg/kg IP) in mice. The anticonvulsant effects of ethanol (1 g/kg IP) were also significantly antagonised by naloxone (1 mg/kg IP) but not by a 0.3 mg/kg IP dose or by cyprodime (3 mg/kg IP). The results with naloxone were confirmed using higher doses of phenytoin (10 mg/kg IP) and ethanol (1.5 g/kg IP). In contrast to the above findings, naloxone (0.3, 1 mg/kg IP) had no effect on the increase in seizure threshold induced by sodium valproate (200 mg/kg IP) or dizocilpine (MK801, 0.5 mg/kg IP) and paradoxically potentiated the increase in seizure threshold produced by phenobarbitone (15 mg/kg IP); carbamazepine (10 mg/kg IP) and the benzodiazepine agonist loprazolam (1 mg/kg IP), clearly differentiating these compounds from phenytoin and ethanol. These findings suggest that the anticonvulsant effects of phenytoin and ethanol (as assessed by their ability to prevent tonic hindlimb extension in the mouse electroshock model) may be mediated, at least in part, by the release of endogenous opioids and subsequent activation of opioid receptors (mu, in the case of phenytoin, but non-mu, in the case of ethanol) although direct activity at opioid receptors cannot be precluded.

Differential development of acute tolerance to the motor impairment and anticonvulsant effects of ethanol.

The development of acute tolerance to the motor impairment and anticonvulsant effects of ethanol was examined. Acute tolerance to the motor impairment effect of ethanol was shown by a decrease in the degree of intoxication, as measured on the moving belt task, at higher blood ethanol levels ranging from 206 to 256 mg/dl. There was no evidence of acute tolerance to the anticonvulsant effect of ethanol in rats tested over the same time period. These results indicate that, like chronic tolerance, acute tolerance to ethanol develops at different rates for different effects of the drug. The fact that chronic tolerance to the anticonvulsant effect of ethanol has been well documented raises doubts about the assumption that similar physiological changes underlie acute and chronic tolerance to a drug effect, and support the idea that the relationship between acute and chronic tolerance is more complex than previously thought.

Amygdaloid kindling and kindled seizures in rats receiving chronic ethanol administration

The effects of chronic ethanol administration were studied in rats receiving amygdaloid kindling. Daily ethanol administration 10 min prior to kindling stimulation delayed acquisition of kindling without affecting the electrical afterdischarge. For the lowest tested dose of ethanol (0.5 g/kg), this delay was restricted to kindling stages 1 and 2. For the higher doses of ethanol (1.0 and 1.5 g/kg) this delay became more severe and stages 3 and 4 were blocked. Ethanol produced a clear dose-related anticonvulsant effect upon kindled seizures. After repeated exposure to kindling stimulation and ethanol this anticonvulsant effect vanished. After a 15-day interval without stimulation or ethanol application kindled animals were insensitive to ethanol's anticonvulsant effect. In conclusion, it is suggested that the anticonvulsant effects of low ethanol doses are restricted to kindling stages 1 and 2 and that anticonvulsant effects of high ethanol doses are limited by tolerance and by the level of consolidation of the kindled seizure. Finally, we suggest that the anticonvulsant properties of ethanol are not due to its general depressant effect but to some rather specific action.

Sensitivity to the anticonvulsant effects of ethanol and pentobarbital in mouse lines genetically selected for ethanol sensitivity.

Mouse lines genetically selected for susceptibility [long sleep (LS)] or resistance [short sleep (SS)] to the acute hypnotic effects of ethanol were tested for sensitivity to maximal electroshock seizures. LS mice were slightly more sensitive than SS mice. Ethanol or pentobarbital pretreatment elevated seizure thresholds in both lines. LS and SS mice were approximately equally protected by ethanol, but LS mice were somewhat more protected than SS mice by pentobarbital. These studies do not provide evidence that sensitivity to the anticonvulsant effect of ethanol is mediated by substantially the same genes as those mediating sensitivity to EtOH's hypnotic effects. However, sensitivity to pentobarbital's anticonvulsant effects may be genetically correlated.

RO15-4513 but not FG-7142 reverses anticonvulsant effects of ethanol against bicuculline- and picrotoxin-induced convulsions in rats.

The reversal of anticonvulsant effect of ethanol against chemoconvulsions by RO15-4513 was investigated in rats as this novel imidazobenzodiazepine (ethyl-8 azido-5, 6-dihydro-5-methyl-6-Oxo-4H-imidazo [1,5a] [1,4] benzodiazepine-3-carboxylate) is reported to antagonize the acute behavioral and biochemical effects of ethanol in animals. Reversal of ethanol effects on onset of myoclonic jerks, tonic extensor phase, mortality time and percent protection against mortality were compared with not only other anticonvulsant pentobarbital but also with another inverse agonist FG-7142. Pretreatment with RO15-4513 (4 mg/kg) reversed the protective effect of ethanol against bicuculline-induced tonic extensor phase and mortality (87%). This response was sensitive to reversal by RO15-1788 (10 mg/kg). However, onset of myoclonic jerks and duration of clonus were not significantly altered. It also reversed the effect against picrotoxin but the reversal against mortality was up to 50%. As compared to ethanol, RO15-4513 reversed partially the protective effect of pentobarbital against bicuculline- and picrotoxin-induced convulsions. FG-7142 failed to reverse the protective effect of ethanol and pentobarbital against bicuculline-induced tonic extensor phase although it reversed the effect against onset and mortality. It had no effect on the protective effect against picrotoxin-induced convulsions. Both RO15-4513 and FG-7142 possessed proconvulsant effects against bicuculline but not against picrotoxin. These observations suggest that RO15-4513 has a more preferential action against ethanol effects as compared to the other inverse agonist.