Ontogenetic development of sensitivity of the cerebral cortex to an antagonist of GABA(A) receptor bicuculline.

Local application of four concentrations of bicuculline methiodide (a specific antagonist of GABA(A) receptors) was used to study a sensitivity of somatosensory cortex in four age groups of immature rats with implanted electrodes. Presence and latencies of two epileptic phenomena (focal discharges and seizures) were evaluated. Focal discharges exhibited moderate tendency to a decrease of sensitivity to bicuculline methiodide with maturation. Concentration-effect relation of incidence of focal discharges was observed only in 7- and 12-day-old but not in older animals. Results with incidence and latencies of seizures did not show relations to age or concentration of bicuculline. Neither of the epileptic phenomena can be used as a reliable index of cortical maturation.

Effect of prenatal methamphetamine exposure and challenge dose of the same drug in adulthood on epileptiform activity induced by electrical stimulation in female rats.

Our previous study demonstrated that chronic prenatal methamphetamine (MA) exposure and a single dose of MA in adulthood decrease focally induced epileptiform activity in adult male rats. As seizures are known to be dependent on sex and female estrous cycle, the goal of the present study was to examine the combined effect of prenatal MA exposure (5mg/kg) and the MA challenge dose (1mg/kg) in adulthood on electroencephalography (EEG) recordings and consequences of brain stimulation in freely moving adult female rats with respect to the estrous cycle. Overall, 12 groups of adult female rats were tested: prenatally MA-exposed, prenatally saline-exposed and rats without prenatal injections, each of these groups was either postnatally challenged with MA or with saline injection (MA-MA, MA-S; S-MA, S-S; C-MA, C-S) and further divided according to the stage of the estrous cycle to metestrus/diestrus (M/D) or proestrus/estrus (P/E). Seizures were induced by repetitive electrical stimulation (15s/8Hz) of sensorimotor cortex. Stimulation threshold, duration of afterdischarges (ADs), and presence and duration of spontaneous ADs (SADs) were evaluated. Additionally, behavior associated with stimulation and ADs, and occurrence of wet-dog-shakes (WDS) were analyzed. The present study demonstrates that the prenatal MA exposure decreased the seizure threshold in females in M/D, but not in females in P/E. In addition, prenatally MA-exposed M/D females injected with saline in adulthood had increased the duration of ADs as well as SADs. The challenge dose of MA also decreased the seizure threshold. Moreover, prenatal as well as adult MA administration decreased the number and occurrence of WDS, respectively. Thus, the present study demonstrates that the effect of prenatal MA exposure and challenge dose of the same drug on focally induced epileptiform activity in adult female rats depends on the estrous cycle.

An antagonist of GABA-B receptors potentiates activity of cortical epileptic foci.

Cortical epileptic foci elicited by local application of bicuculline methiodide represent a model of interictal epileptic activity with a transition into ictal phases. We studied a role of GABA-B receptors in this model using GABA-B receptor antagonist CGP35348 in adult rats with implanted cortical electrodes and cannula. CGP35348 (100 or 200 mg/kg i.p.) did not affect interictal discharges but it augmented ictal activity. Latency to the first ictal episode was decreased by the lower dose of CGP35348, duration of episodes was increased by the higher dose. GABA-B receptor antagonist did not influence purely cortical epileptic phenomenon but it is proconvulsant in ictal activity generated with participation of subcortical structures.

Increased seizure susceptibility induced by prenatal methamphetamine exposure in adult female rats is not affected by early postnatal cross-fostering.

Our previous studies repeatedly demonstrated that prenatal methamphetamine (MA) exposure alters seizure susceptibility in adult rats. Both the inhibitory GABA system and the excitatory NMDA system play a role in the effect of MA on epileptic seizures. On the basis of our previous behavioral results, the effect of cross-fostering on seizure susceptibility in adult female rats was examined in the present study. Bicuculline (GABA(A) receptor antagonist) and NMDA (NMDA receptor agonist) were used to induce seizures in adult female offspring exposed to MA in the prenatal and/or preweaning periods. Female dams were injected with MA (5mg/kg daily) or physiological saline (S) for approximately 9 weeks [about 3 weeks prior to impregnation, for the entire gestation period (22 days), and in the preweaning period (21 days)]. Absolute controls (C) did not receive any injections. On postnatal day 1, pups were cross-fostered so that each mother received pups from all three treatments. Thus, nine groups (based on the prenatal and postnatal drug exposures) of adult female rats were tested in each seizure test: C/C, C/S, C/MA, S/C, S/S, S/MA, MA/C, MA/S, MA/MA. The present study demonstrated that both the excitatory NMDA system and the inhibitory GABA system are involved in the proconvulsive effect of MA during prenatal and partially also postnatal development in female rats. However, because our results did not show any improvement in seizure susceptibility in prenatally MA-exposed animals that were fostered by control mothers (MA/C) relative to their siblings fostered by MA-treated mothers (MA/MA), our hypothesis of the cross-fostering effect seems to be incorrect in contrast to our behavioral studies.

Effect of cross-fostering on seizures in adult male offspring of methamphetamine-treated rat mothers.

Stimulant drugs are often associated with increased seizure susceptibility. Inhibitory gamma-aminobutyric acid (GABA) and excitatory N-methyl-D-aspartate (NMDA) systems play a role in the effect of stimulants in the genesis of epileptic seizures. Our previous studies showed that prenatal methamphetamine (MA) exposure induced long-term changes in seizure susceptibility. The aim of the present study was to investigate the effect of cross-fostering on the prenatal and postnatal MA-exposed rats, respectively, on their seizures in adulthood. Bicuculline (GABA(A) receptor antagonist), NMDA (NMDA receptor agonist) and flurothyl (a convulsant gas) were used to induce seizures in adult male offsprings. Female dams were injected with MA (5 mg/kg daily) or physiological saline (S) for approx. 9 week [about 3 week prior to impregnation, for the entire gestation period (22 days) and in preweaning period (21 days)]. Absolute controls (C) did not receive any injections. On postnatal day 1, pups were cross-fostered so that each mother received pups from all three treatments. Thus, nine groups (based on the prenatal and postnatal drug exposure) of adult male rats were tested in each seizure test: C/C; C/S; C/MA; S/C; S/S; S/MA; MA/C; MA/S; MA/MA. The present study demonstrates that the effect of prenatal and/or postnatal MA exposure is seizure model specific. In addition, our data show that there is an effect of cross-fostering on seizures; particularly, the effect of prenatal MA exposure shown in animals fostered by control mothers is no longer apparent in animals fostered postnatally by MA-treated mothers. Such effect of postnatal treatment is not manifested in prenatal controls. In summary, it seems that: (1) prenatal MA exposure alters seizure susceptibility more than postnatal MA exposure; (2) especially in seizures induced by chemicals that affect GABAergic system (bicuculline, flurothyl) notable effect of adoption (cross-fostering) is apparent; (3) in seizure models that are associated with NMDA system (NMDA, flurothyl), effect of prenatal stress seems to play a role.

Similar effects of lamotrigine and phenytoin against cortical epileptic foci in immature rats.

Action of antiepileptic drugs in immature brain may differ from that in adult brain. The aim of our study was to study an anticonvulsant action of lamotrigine and phenytoin, i.e. two drugs active against partial seizures in adult experimental animals as well as human patients, in a model of simple partial seizures in immature rats. Epileptic foci were induced by local application of bicuculline methiodide on sensorimotor cortical area of 12-day-old rat pups. The animals were pretreated with lamotrigine (LTG, 10 or 20 mg/kg i.p.) or phenytoin (PHT, 15, 30 or 60 mg/kg i.p.). Control rats for LTG received saline, controls for PHT solvent composed of propyleneglycol, ethanol and water. Influence of either drug on interictal activity was negligible. High doses of both LTG and PHT suppressed the transition into ictal phases and shortened the duration of persisting seizures. The tricomponent solvent exhibited moderate activity against ictal activity if compared with saline controls. The two drugs exhibited similar action in our model, i.e. the suppression of secondary generalization from epileptic focus. This action is comparable to that described for human patients and adult experimental animals. In favor of lamotrigine speaks the absence of serious side effects.

Bone metabolism: a note on the significance of mouse models.

This minireview briefly surveys the complexity of regulations governing the bone metabolism. The impact of clinical studies devoted to osteoporosis is briefly summarized and the emphasis is put on the significance of experimental mouse models based on an extensive use of genetically modified animals. Despite possible arising drawbacks, the studies in mice are of prime importance for expanding our knowledge on bone metabolism. With respect to human physiology and medicine, one should be always aware of possible limitations as the experimental results may not be, or may be only to some extent, transposed to humans. If applicable to humans, results obtained in mice provide new clues for assessing unforeseen treatment strategies for patients. A recent publication representing in our opinion the important breakthrough in the field of bone metabolism in mice is commented in detail. It provides an evidence that skeleton is endocrine organ that affects energy metabolism and osteocalcin, a protein specifically synthesized and secreted by osteoblasts, is a hormone involved. If confirmed by other groups and applicable to humans, this study provides the awaited connection of long duration between bone disorders on one hand and obesity and diabetes on the other.

Influence of melatonin pretreatment and preconditioning by hypobaric hypoxia on the development of cortical photothrombotic ischemic lesion.

Photothrombotic model of ischemia (PT) is based on free radical-mediated endothelial dysfunction followed by thrombosis. Free radicals are also involved in hypoxic preconditioning. We tested the sensitivity of PT to preconditioning with hypobaric hypoxia and to pretreatment with melatonin. In adult Wistar rats, after intravenous application of Rose Bengal, a stereo-tactically defined spot on the denuded skull was irradiated by a laser for 9 min. The first experimental group underwent hypobaric hypoxia three days before irradiation. In the second experimental group, melatonin was applied intraperitoneally one hour before irradiation. Three days after irradiation, animals were sacrificed, the brains perfused, and stained with TTC. Ischemic lesions were divided into grades (I, II, III). In the control group (where no manipulation preceded photothrombosis), most animals displayed deep damage involving the striatum (grade III). The group pre-exposed to hypoxia showed similar results. Only 28.57 % of the melatonin pretreated animals exhibited grade III lesions, and in 57.14 % no signs of lesions were detected. Pre-exposure to hypoxia was not protective in our model. Pretreatment with melatonin lead to a significant reduction of the number of large ischemic lesions. This result is probably caused by protection of endothelial cells by melatonin.

Long-term changes of activity of cortical neurons after status epilepticus induced at early developmental stages in rats.

Spontaneous activity of cortical neurons was studied under urethane anesthesia in adult rats 3 months after convulsive status epilepticus induced by lithium-pilocarpine administration at the age of 12 (SE12 group) or 25 (SE25 group) days. Whereas random firing neurons dominated in control animals (61 out of 98 cells), SE25 animals exhibited a significant increase in the incidence of bursting cells (38 out of 59 units). Similar change in SE12 animals did not reach the level of statistical significance. Status epilepticus at an early developmental stage may result in a long-lasting change in functions of surviving cortical neurons.

Action of GABA-B antagonist on cortical epileptic afterdischarges in rats is similar to that of GABA-A antagonist.

Threshold intensities for epileptic phenomena induced by cortical stimulation were used for comparison of the action of GABA-B and GABA-A antagonists in rats with implanted electrodes. Both CGP 35348 (200 mg/kg i.p.) and bicuculline (4 mg/kg i.p.) significantly decreased thresholds for spike-and-wave afterdischarges and their motor counterpart (clonic seizures) whilst transition into the second, limbic type of afterdischarge as well as threshold for movements directly bound to stimulation remained uninfluenced by either drug.

Proconvulsant effect of aminophylline on cortical epileptic afterdischarges varies during ontogeny.

Effect of aminophylline on epileptic afterdischarges (ADs) induced repeatedly by rhythmic electrical stimulation of sensorimotor cortical area was studied in rat pups 12, 18 and 25 days old. The proconvulsant effect of aminophylline (50 and/or 100 mg/kg i.p.) was more expressed in 12- and 18-day-old rats than in the oldest group. In 12-day-old rat pups there was an enormous increase of transition of the spike-and-wave type of ADs into the second, limbic type, a situation observed only exceptionally under control conditions. A prolongation of ADs was related to this transition (limbic ADs are always longer than spike-and-wave ones). Eighteen-day-old rats exhibit this transition less frequently but a marked prolongation of spike-and-wave ADs was recorded in a part of these animals forming a pattern of status lasting some tens of minutes. Aminophylline led only to a transient prolongation of spike-and-wave ADs in the oldest group. The transition into the limbic type of ADs was seen in this age group only exceptionally what is in contrast to age-matched controls in which this transition is common. The effect of aminophylline on cortical ADs which is most marked in the youngest group changes qualitatively during postnatal development.

GABA uptake blocker NNC-711 exhibits marked anticonvulsant action in two cortical epileptic models in immature rats.

PURPOSE: To study the anticonvulsant action of a gamma-aminobutyric acid (GABA) uptake inhibitor NNC-711 in two models of cortical epileptogenesis in immature rats. METHODS: Twelve-, 18-, and 25-day-old rat pups with implanted electrodes were used in this study. Epileptogenic foci were elicited by a local application of bicuculline methiodide (BMI) on the sensorimotor cortical region by means of an implanted cannula, and cortical epileptic afterdischarges (ADs) were induced by low-frequency stimulation (8 Hz) of the same cortical area. Epileptogenic foci were formed after pretreatment with NNC-711 (1 or 10 mg/kg, i.p.), and epileptic ADs were elicited in the second experimental series. Then NNC-711 was administered in the same doses, and stimulation was repeated. RESULTS: NNC-711 did not block the formation of epileptogenic foci, but it significantly suppressed the spontaneous transition of interictal focal into ictal activity in all age groups. The intensity of movements accompanying stimulation of the sensorimotor cortex was less under the influence of NNC-711 in the 18- and 25-day-old rats. The duration of cortical ADs was shortened in all age groups, but transient abolition of ADs was observed only after the higher dose in the 25-day-old rats. In addition, the intensity of clonic seizures appearing during ADs decreased, and the transition of ADs into another type due to an involvement of limbic structures failed to appear in the 18- and 25-day-old rats. CONCLUSIONS: Primary epileptogenesis in the cerebral cortex was hardly influenced by NNC-711, but the spread of epileptic activity was markedly suppressed. This effect was better expressed in the 18- and 25-day-old animals than in the youngest group.

Motor and electrocorticographic epileptic activity induced by bicuculline in developing rats.

Motor seizures were induced by intraperitoneally (i.p.) injected bicuculline in 270 rats aged 7, 12, 18, 25, or 90 days. Bicuculline was able to elicit both minimal (clonic) and major (tonic-clonic) seizures in all age groups, but in 7-day-old rats minimal seizures were only noted exceptionally. CD50s (for major seizures) ranged from 2.48 to 2.85 mg/kg in the three younger groups and increased to approximately 7 mg/kg in 25- and 90-day-old rats. An intravenous (i.v.) administration of bicuculline in 67 rats, 18 and 25 days old, caused identical CD50s in these groups, indicating that the difference that occurs with an i.p. administration is due to pharmacokinetic reasons. Electrocorticographic (ECoG) studies in acute experiments as well as in young rats with implanted electrodes demonstrated general principles of the development of EEG: an increase in frequency of individual elements, in generalization of the epileptic activity, in synchronization of activity among various cortical regions, and in the correlation between ECoG and motor phenomena. An exception occurred as an age-related phenomenon: rhythmic activity of the spike-and-wave type. This activity appeared in 18-day-old and older rats and was invariably accompanied by "freezing" of the animals.

Influence of cinromide on metrazol--induced seizures during ontogenesis in rats.

The effects of cinromide on two types of metrazol-induced seizures were studied in 236 male rats aged 7, 12, 18, 25 and 90 days. Cinromide was administered intraperitoneally in a dose of 25 or 50 mg.kg-1 30 min before a subcutaneous injection of metrazol. Major, i.e. generalized tonic-clonic seizures could be elicited by metrazol in all age groups. Cinromide was efficient against this type of seizures at all developmental stages, but its action in 7- and especially in 12-day-old rat pups was less marked than in older animals. Minimal metrazol seizures (predominantly clonic) could be reliably elicited since the age of 18 days. Cinromide was also able to suppress this type of seizures, but in adult rats the dose of 75 mg.kg-1 had to be administered because even the 50 mg.kg-1 dose was not sufficient. Quantitative changes of the antimetrazol action of cinromide were demonstrated during maturation in rats.

Ontogenetic development of convulsant action of Ro 5-3663 in the rat.

Motor seizures were induced by Ro 5-3663 in 156 male albino rats aged 7,12,18,25, and 90 days. Both minimal and maximal seizures could be elicited in 18-day-old and older animals, whereas only maximal seizures were induced in the two youngest groups. ECoG changes were studied in other 21 young rats. First changes induced by Ro 5-3663 were formed by isolated sharp waves in 7- and 12-day-old rats and by episodes of rhythmic activity in older animals. An imperfect form of this rhythmic activity could be seen even in 12-day-old rats. Ictal ECoG activity exhibited an increase in frequency of individual graphoelements, in generalization and in synchronization of activity among different cortical regions with maturation. Correlation between motor and ECoG phenomena was poor in 7-day-old rats and ameliorated with age but it never reached perfection. The actions of Ro 5-3663 are identical with those induced by metrazol but they differ from those elicited by bicuculline or 3-mercaptopropionic acid.

Convulsant action of diphenylhydantoin overdose in young rats.

Acute toxicity of diphenylhydantoin (DPH) was studied in 241 male albino rats aged 7, 12, 18, 25 and 90 days. Single intraperitoneal dose of DPH (from 200 to 1000 mg/kg) induced only ataxia and loss of righting reflex in 25-day-old and adult rats. In rats aged 18 days or less ataxia of hindlimbs was also marked. In all these age groups generalized convulsions appeared; they were formed by wild running followed by a clonic phase. The dose of DPH necessary for elicitation of seizures was lowest in 7-day-old rats (75 mg/kg) and increased with age up to 200 mg/kg in 18-day-old rats. The 1000 mg/kg dose was lethal for 25- and 12-day-old rats, but not for 7-day-old ones. The uneven development of excitatory and inhibitory action of DPH is suggested.