Susan M. Irtenkauf

Central anticonvulsant effects of magnesium sulfate on N-methyl-D-aspartate-induced seizures

OBJECTIVE The objective of this study was to determine if magnesium sulfates central anticonvulsant activity is effective against N-methyl-D-aspartate-induced seizures. STUDY DESIGN In two separate experiments we investigated magnesium sulfates ability to inhibit N-methyl-D-aspartate-induced hippocampal seizures in rats. In the first experiment magnesium sulfate was administered peripherally before an intracranial injection of 20 micrograms of N-methyl-D-aspartate. In the second experiment magnesium sulfate was injected intracranially concurrently with N-methyl-D-aspartate. The ability of magnesium sulfate to suppress N-methyl-D-aspartate-induced seizure activity under both conditions was assessed. RESULTS Peripherally administered magnesium sulfate significantly increased the latency from the time of an N-methyl-D-aspartate injection to the first seizure both by acute injection and after 2 hours of sustained elevation of serum magnesium levels when compared with saline solution-injected controls (p < 0.01). The duration of the first seizure was also significantly reduced. Intracranially administered magnesium sulfate significantly (p < 0.01) increased the seizure latency period by 120%. Overall, central magnesium sulfate prevented seizure activity in 40% of the animals (p < 0.01). CONCLUSION Magnesium sulfate has a central anticonvulsant action on N-methyl-D-aspartate-induced seizures in this rat model of hippocampal seizures.

Peripheral magnesium sulfate enters the brain and increases the threshold for hippocampal seizures in rats

OBJECTIVES Our objectives were to determine whether magnesium sulfate crosses the blood-brain barrier and whether it has central anticonvulsant action. STUDY DESIGN In experiment 1 34 female Long-Evans rats were divided into six groups: control (n = 7); single magnesium sulfate injection and evaluation after 20 minutes in 3 conditions: normal rats (n = 7), sham-operated animals (n = 5), and after electrical stimulation by hippocampal electrode (n = 5); single injection and evaluation after 2 hours (n = 5); and prolonged (2 hours) serum magnesium elevation (n = 5). Serum, cerebrospinal fluid, and specific brain areas were analyzed for magnesium concentrations. In experiment 2 threshold for electrical seizure was measured in eight rats before and after intraperitoneal injections of magnesium sulfate versus saline solution. RESULTS In experiment 1 there was a significant correlation between blood and cerebrospinal fluid magnesium concentrations (r = 0.80, p < 0.0001). Magnesium concentrations were increased in the cortex and hippocampus, with the largest changes occurring after two hours of sustained serum magnesium concentrations (p < 0.01). Induction of hippocampal seizure activity resulted in further elevations in cerebrospinal fluid magnesium concentrations but did not change brain concentrations. In experiment 2 magnesium sulfate increased the electrical threshold required to induce seizures by 34% (p = 0.01). CONCLUSIONS Magnesium sulfate enters the cerebrospinal fluid and brain and has a central anticonvulsant effect.

Magnesium Sulfate Treatment Decreases N-Methyl-D-Aspartate Receptor Binding in the Rat Brain: An Autoradiographic Study

OBJECTIVE: We determined the effect of peripherally administered magnesium sulfate on N-methyl-D-aspartate (NMDA) receptor binding capacity in various regions of the rat brain. METHODS: Three separate experiments were performed. 1) Six rats were injected intraperi toneally with 270 mg/kg of magnesium sulfate, followed by 27 mg/kg every 20 minutes for 4 hours; controls (n = 6) received saline. 2) Six rats received intraperitoneal injections of magne sium sulfate (270 mg/kg) every 4 hours for 24 hours, while six received saline. 3) Six rats received intraperitoneal magnesium sulfate (270 mg/kg) every 12 hours for a total of 2 weeks, and six received saline. Rats were subsequently perfused and sacrificed, and their brains were dissected, rinsed, and frozen. Cryostat sections were taken, labeled by in vitro [3H]-CGP 39653, assayed autoradiographically, and mounted on Ultrofilm for 4 weeks. Optical density measurements of binding on each section were performed using an image analyzing system. Eleven brain regions were sampled: 1, 2) frontal and occipital cortex; 3-7) hippocampus—CA-1, CA-3, stratum radiatum, stratum oriens, dentate gyrus; 8) thalamus; 9) hypothalamus; 10) caudate nucleus; and 11) cerebellum. RESULTS: The NMDA receptor binding density in the hippocampus was significantly higher than in all other brain regions in all three experiments. In experiment 1, there was no significant effect on NMDA receptor binding. However, prolonged systemic administration of magnesium sulfate for 24 hours resulted in significantly reduced [3H]-CGP binding in all brain regions sampled. After chronic magnesium sulfate administration (2 weeks), the [3H]-CGP binding was still reduced in the cortex and some regions of the hippocampus; however, there was no significant change in other regions. CONCLUSIONS: Peripheral treatment with magnesium sulfate results in a significant reduction in the NMDA receptor binding capacity in the rat brain. These results support the hypothesis that magnesium central activity is mediated, at least in part, via the NMDA receptor. (J Soc Gynecol Invest 1994 ;1 :25-30)

Neurochemical changes after acute binge toluene inhalation in adolescent and adult rats: A high-resolution magnetic resonance spectroscopy study

Inhalant abuse in young people is a growing public health concern. We reported previously that acute toluene intoxication in young rats, using a pattern of exposures that approximate abuse patterns of inhalant use in humans, significantly altered neurochemical measures in select brain regions. In this study, adolescent and young adult rats were exposed similarly to an acute (2 x 15 min), high dose (8000-12,000 ppm) of toluene and high-resolution magic angle spinning proton magnetic resonance spectroscopy (HR-MAS 1H-MRS) was used to assess neurochemical profiles of tissue samples from a number of brain regions collected immediately following solvent exposure. The current investigation focused on N-acetyl-aspartate (NAA), choline-containing compounds, creatine, glutamate, GABA, and glutamine. Contrary to our predictions, no significant alterations were found in the levels of NAA, choline, creatine, glutamate, or glutamine in adolescent animals. In contrast to these minimal effects in adolescents, binge toluene exposure altered several neurochemical parameters in young adult rats, including decreased levels of choline and GABA in the frontal cortex and striatum and lowered glutamine and NAA levels in the frontal cortex. One of the more robust findings was a wide-ranging increase in lactate after toluene exposure in adult animals, an effect not observed in adolescents. These age-dependent effects of toluene are distinct from those reported previously in juvenile rats and suggest a developmental difference in vulnerability to the effects of inhalants. Specifically, the results suggest that the neurochemical response to toluene in adolescents is attenuated compared to adults, and imply an association between these neurochemical differences and age-influenced differences in solvent abuse in humans.

Magnesium is more efficacious than phenytoin in reducing N-methyl-D-aspartate seizures in rats

OBJECTIVE Although magnesium sulfate is one of the most commonly used agents for seizure prophylaxis in preeclampsia, its efficacy relative to other anticonvulsants is incompletely investigated. The underlying mechanisms of eclamptic seizures are unknown, and there is currently no universally accepted animal model for eclampsia. However, one commonly used method for studying the relative efficacy of antiepileptic drugs is through their effect on N-methyl-D-aspartate-induced seizures. Our aim was to compare the anticonvulsant effects of phenytoin and magnesium sulfate in an N-methyl-D-aspartate-induced seizure model. STUDY DESIGN Twenty-one female rats were each stereotaxically implanted with a chronic indwelling bipolar recording electrode in the hippocampus and an injection cannula in the lateral cerebral ventricle. After 7 days animals were randomly given 90 mg/kg magnesium sulfate (n = 7), 50 mg/kg phenytoin, or saline solution (n = 7) intravenously. Fifteen minutes after the infusions animals were given 20 micrograms/microliters N-methyl-D-aspartate by direct intraventricular injection, and seizure activity was assessed for 20 minutes thereafter. All data were analyzed with the Mann-Whitney test. RESULTS When compared with saline solution controls, total duration of seizure activity in animals treated with magnesium sulfate was significantly decreased (p < 0.05) and time to onset of seizure activity was significantly increased (p < 0.05). However, rats that received phenytoin did not show significant changes in these parameters. The post-N-methyl-D-aspartate seizure mortality rate was 50% in the saline solution controls and 29% in the phenytoin group, whereas none of the rats that received magnesium sulfate died. CONCLUSION These results suggest that magnesium sulfate is a significantly more effective prophylactic agent than phenytoin for N-methyl-D-aspartate-induced seizures.

Comparison of toluene-induced locomotor activity in Four Mouse Strains

The mechanisms by which abused inhalants exert their neurobehavioral effects are only partially understood. In research with other drugs of abuse, specific inbred mouse strains have been useful in exploring genetic loci important to variation in behavioral reactions to these drugs. In the present investigation, mice from three inbred strains (Balb/cByj, C57BL/6J and DBA/2J) and one outbred strain (Swiss Webster) were studied for their acute and chronic sensitivity to toluene-induced changes in locomotor activity. Mice were exposed to toluene (0, 100, 2000, 8000, and 10,000 ppm) for 30 min in static exposure chambers equipped with activity monitors. In the acute condition, concentrations of toluene <8000 ppm increased ambulatory distance while the concentrations of > or =8000 ppm induced temporally biphasic effects with initial increases in activity followed by hypoactivity. Between-group differences in absolute locomotor activity levels were evident. The inbred Balb/cByj and DBA/2J strains as well as the outbred Swiss Webster strain of mice showed greater increases in activity after an acute challenge exposure to 2000 ppm than the inbred C57BL/6J strain. The same animals were then exposed 30 min/day to 8000 ppm toluene for 14 consecutive days. Re-determination of responses to 2000-ppm challenge exposures revealed that sensitization developed in locomotor activity and that the DBA/2J strain showed the greatest increase in sensitivity. These baseline differences in acute sensitivity and the differential shifts in sensitivity after repeated exposures among the inbred mouse strains suggest a genetic basis for the behavioral effects to toluene. The results support the notion that like for other drugs of abuse, using various strains of mice may be useful for investigating mechanisms that underlie risk for inhalant abuse.

Stimulation and inhibition of N-methyl-D-aspartate receptors in rats: Developing a seizure model

OBJECTIVE The objective of this study was to develop an experimental rat hippocampal seizure model based on the stimulatory effects of N-methyl-D-aspartate and to determine the inhibitory effects of MK-801 on N-methyl-D-aspartate-induced seizures. STUDY DESIGN Two separate experiments were performed. In the first experiment chemitrode-implanted rats were injected intracranially with increasing doses (5, 10, 20, and 30 micrograms) of N-methyl-D-aspartate into the hippocampus. Various electrophysiologic and behavioral parameters were examined to determine the dose required to reliably elicit hippocampal seizure activity without having toxic effects on the rats. In the second experiment rats were given an intraperitoneal injection of MK-801 (0.5 or 1 mg/kg), followed 20 minutes later by an intracranial injection of N-methyl-D-aspartate (20 or 30 micrograms). The ability of MK-801 to suppress N-methyl-D-aspartate-induced seizure activity was assessed in this experiment. RESULTS Intrahippocampal injection of 20 micrograms of N-methyl-D-aspartate produced the shortest electrical seizure latency (193 +/- 72 seconds, p < 0.01). At this dose seizure was achieved in 80% (four of five of the animals, and the highest numbers of electrical seizures per animal were produced (2.2 +/- 0.8, p < 0.05). The group that received 30 micrograms of N-methyl-D-aspartate had a shorter latency, a longer duration of behavioral seizure and a higher number of behavioral seizures (p < 0.05). However, this group suffered a 60% (three of five) mortality rate. The addition of MK-801 significantly decreased the number of seizures per animal and the total seizure duration (p < 0.05). MK-801 also reduced the latency period. CONCLUSION Intracranial injection of 20 micrograms of N-methyl-D-aspartate produced reliable hippocampal seizure activity without mortality. MK-801 at a dose of 1 mg/kg injected intraperitoneally had significant inhibitory effects on this seizure model.

Anticonvulsant effects of magnesium sulfate in hippocampal-kindled rats

The objective of the present study was to determine whether magnesium sulfate has anticonvulsant actions in the hippocampal-kindled rat model of epilepsy. Fully kindled rats received acute intraperitoneal injections of magnesium sulfate (270 mg/kg), phenytoin (20 mg/kg) or saline in random order. Electrical seizure duration, behavioral seizure stage and duration of postictal EEG depression were examined 15, 30 and 60 min after injection. In an additional group of rats, kindled seizures were measured before and after chronic (2 h) intraperitoneal injections of magnesium sulfate versus saline. There was a significant decrease in electrical seizure duration (p < 0.01) and behavioral seizure stage (p < 0.01) with acute magnesium sulfate injections compared to saline injections. Phenytoin had no statistically significant effects on hippocampal-kindled seizures. Chronic magnesium sulfate treatment significantly reduced behavioral seizure stage at 2, 24, and 48 h postinjection (p < 0.05), but did not affect seizure duration. There was a significant time by treatment effect for magnesium sulfate on postictal EEG depression (p < 0.01). We conclude that in this model of hippocampal epilepsy-induced (kindled) rats, magnesium sulfate has significant anticonvulsant effects. Copyright 1995 S. Karger AG, Basel

Maternal Electrical Seizures Are Associated with an Increase in Perinatal Death in Rat Pups

Relatively few investigators have examined the effect of pregnancy on seizure susceptibility. The present study was designed to determine whether there are any changes in hippocampal seizure thresholds during different stages of pregnancy. Second, we examined the effect of maternal seizures on perinatal mortality.Sixteen female rats were surgically anesthetized and a bipolar electrode stereotaxically implanted into the dorsal hippocampus. Following 1-week recovery, baseline values for seizure threshold were obtained for all rats. Seizure thresholds were then re-determined on days 7, 14, and 21 from baseline. Rats were then mated, and seizure thresholds again re-determined at days 7, 14, and 21 of pregnancy. Vaginal smears were taken 3 times weekly to determine stage of estrus on days stimulated during nonpregnancy. The amount of electrical current required to initiate a hippocampal seizure significantly decreased across each of the three stimulations before pregnancy (P <. 01). Vaginal smears indicated th...

Serum ionized magnesium levels decrease with gestational age in normal pregnant women

193 SERUM IONIZED MAGNESIUM LEVELS DECREASE WITH GESTATIONAL AGE IN NORMAL PREGNANT WOMEN. ~ , J.E. Whitty, B.A. Mason, S.M. Irtenkauf*, D.B. Cotton. Department of Ob/Gya, Hutzel Hospital, Wayne State University. OBJECTIVE: Longitudinal changes in serum ionized magnesium. (IMg) and total magnesium (TMg) during pregnancy have not been adequately described. The aim of the present study was to determine levels of IMg, TMg, ionized calcium (ICa), sodium (NA), potassium (K) and pH values over the course of pregnancy in normal pregnant women. STUDY DESIGN: We obtained venous serum samples from 27 normal pregnant women during their first, second and third trimesters. Gestational ages ranged from 6-37 weeks. Samples were analyzed for IMg, TMg, ICa, Na, K and pH using a biomedical stat profile chemistry analyzer with specific ion sensitive electrodes. Data were analyzed with repeated measures ANOVA. RESULTS: In normal pregnant patients, beth serum IMg and TMg decreased linearly with increasing gestational age (p TERB > RITO. CONCLUSION: Similar K~ values and receptor responses support the use of mononuclear leukocytes as a model for I~adrenoreceptors in less accessible myometrium. (Supported by the ETSU Research Development Committee).