Increase in the activity of an epileptogenic focus in frog hippocampus as a result of kynurenines and its reduction by serotonin]

In frogs with epileptogenic focus induced by injection of penicillin (1000 U in 0.4 microliter) into the primordial hippocampus it was shown that pretreatment with two kynurenines (quinolinic acid -- 0.1 microgram, and d,l-kynurenine -- 1 microgram) into the focus region and their injection into the functioning epileptogenic focus led to a sharp increase of the interparoxysmal epileptiform discharges and electrographic correlates of the fit on the EEG. Anthranilic acid (5 microgram) did not influence the activity of epileptogenic foci, and serotonin (1 microgram) and 5-methoxytryptamine (1 microgram) decreased it significantly. It is suggested that the effect of kynurenines on neurones in the epileptogenic foci may play a certain role in the pathogenesis of epilepsy.     

A specific 70K protein found in epileptic rat cortex: induction of bursting activity and negative resistance by its intracellular application in Euhadra neurons

The effect of 70-kD protein (P70, a specific protein found in cobalt-induced epileptogenic focus of rat cerebral cortex) on membrane properties was examined in identified neurons of the snail, Euhadra peliomphala, using the pressure injection method combined with the voltage-clamp technique. In neurons that normally exhibited spontaneous regular firing, intracellular injection of P70 elicited bursting activity and a negative slope resistance (NSR) region in their current-voltage (I-V) curve in a manner corresponding to the duration of its injection. These responses were suppressed by prior injection of an antibody to P70 into the neurons, and were markedly inhibited by a reduction of extracellular Na+ ions and the anticonvulsant agent phenytoin, but not by Co(2+)-substituted Ca(2+)-free saline. In addition, intracellularly applied P70 potentiated both bursting activity and the NSR induced by a Na channel activator, veratridine. However, prior application of a saturating dose of this activator occluded the effect of P70. These results suggest that P70 elicits a Na(+)-dependent negative resistance, which may contribute to the generation of bursting activity.     

Pacemaker potentials are the physiologic basis of epileptiform activity in the buccal ganglia of Helix pomatia

Mechanisms of epileptic activity in nervous systems were studied using the identified neurons B1 through B4 in the buccal ganglia of the snail Helix pomatia as a model system. Activities were recorded with intracellular microelectrodes. Epileptiform activity was induced by bath application of an epileptogenic drug (pentylenetetrazol: 1 mM to 40 mM, or etomidate: 0.1 mM to 1.0 mM). Epileptiform potentials recorded from the somata of neurons consisted of paroxysmal depolarization shifts (PDSs). With increasing concentration of an epileptogenic drug, pacemaker potentials in neuron B3 developed into PDS. Simultaneously several types of chemical post-synaptic potentials were suppressed in amplitude. Since on the one hand epileptic seizures only appear when PDS are synchronized in many neurons and since on the other hand synaptic potentials were found to be suppressed during epileptic conditions, mechanisms underlying neuronal synchronization were studied. Evidence was found that, under epileptogenic conditions only, neurons were synchronized by an non-synaptic release of substances. Strong depolarizations accompanied by an increase in intracellular calcium concentration are known to induce an unspecific exocytosis. Thus, an unspecific exocytosis from the dendrites of PDS-generating neurons probably appears under epileptic conditions and synchronizes neighbouring neurons.     

Effect of diazepam on the Na, K-ATPase state in a penicillin--induced hyperactivity focus in the cerebral cortex]

Intramuscular injection of diazepam to rats at doses of 0.01 and 2 mg/kg 25-30 min after penicillin application to the rat brain cortex leads to alteration of periodic appearance of epileptic seizures (ES), to changes in the seizure pattern, and to emergence of periodic acceleration of epileptiform discharges (ED). Injection of diazepam at a dose of 2 mg/kg 20 min before penicillin application results in the reduction of ED latency in the epileptogenic focus and in a decrease in their frequency before seizures as compared to the control animals without diazepam injection. ES appear irregularly, their quantity is markedly reduced while duration is increased. Diazepam injection leads to disappearance of the rat moving reaction during ER and ES. In vivo experiments diazepam (2 mg/kg) does not influence brain cortex Na, K-ATPase of crude synaptosomes. However, diazepam leads to an increase in Na, K-ATPase activity both in the primary and dependent secondary epileptogenic foci. It is suggested that the anticonvulsant action of diazepam may be underlain by its activating effect on Na, K-ATPase of neuronal membranes in the epileptogenic focus.     

脑肿瘤时致癫痫病变的免疫组织化学和电镜观察的研究

11例肿瘤致癫痫病患者在脑皮质电图和深部电极描记监测致癫痫灶下,行肿瘤和致痫灶的切除。用免疫组织化学方法检测肿瘤和致痫灶内的细胞浆内胶质原纤维酸性蛋白、神经细丝、纤维联结蛋白和内皮细胞及电镜观察。发现致痫灶分别在肿瘤灶(5例)和肿瘤的周边区(6例)内,每一致痫灶内见残留神经元或变性的轴索由瘤性或增生的星形细胞的胶质原纤维或增生血管周细胞的胶原原纤维围绕或包绕;致痫灶内神经元固缩或空泡变性,线粒体肿胀、粗面内浆网扩张,轴索变性及髓鞘板层分离。结合上述所见,讨论了星形细胞在癫痫发作中的机理。     

Induction and blockade of epileptic foci by intracerebral injection of glutamatergic agonists and antagonists in frerly moving cats

The aim of the present work was to test in adult cats the capability of three glutamatergic agonists, NMDA, AMPA and ACDP as epileptogenic agents. Drugs were microinjected in amygdala or hippocampus, and once generated an epileptic focus three selective glutamatergic antagonists NMDA, CNQX and MCPG, were tested. Before and after injection both the EEG and the behavior were continuously monitored. The results were as follows: 1) AMPA showed a greater capability than NMDA or ACPD to generate a chronic epileptic focus; 2) AMPA elicited a greater epileptogenic effect in hippocampus than in amygdala; NMDA had similar epileptogenic effect in both cited structures, and ACPD had not effect; 3) of the three glutamatergic antagonists used to block a long lasting focus, the most effective one was CNQX, which showed a greater effect in hippocampus than in amygdala; 4) comparison between the epileptogenic effect of AMPA and kainic acid (first paper) in the same structure, showed that kainic acid is about 15 fold more epileptogenic. A discussion of the probable mechanisms of these results was undertaken.     

Selective Depletion of Dopamine, Octopamine, and 5-Hydroxytryptamine in the Nervous Tissue of the Cockroach (Periplaneta americana)

High-performance liquid chromatography with electrochemical detection was used to measure the concentrations of 3,4-dihydroxyphenylethylamine (dopamine), 5-hydroxytryptamine (5-HT), p-hydroxyphenylethanolamine (octopamine), alpha-methyl-p-tyrosine, and tryptophan in the cerebral ganglia of cockroaches (Periplaneta americana) after peripheral administration of alpha-methyl-p-tyrosine and alpha-methyltryptophan. In addition, the levels of dopamine, 5-HT, octopamine, alpha-methyl-p-tyrosine, and tryptophan were determined after injection of alpha-methyl-p-tyrosine, 6-hydroxydopamine, or 5,7-dihydroxytryptamine directly into the cerebral ganglia by means of microinjection needles. Peripheral administration of alpha-methyl-p-tyrosine (400-1,600 micrograms/insect) caused a reduction in dopamine and 5-HT concentrations in cockroach cerebral ganglia, although the reduction in dopamine concentrations was more pronounced. Peripheral injections of alpha-methyl-p-tyrosine also reduced octopamine levels in the cerebral ganglia. Peripheral injection of alpha-methyltryptophan (400-1,600 micrograms/insect) caused a marked reduction in 5-HT and tryptophan concentrations in cockroach cerebral ganglia without altering dopamine or octopamine concentrations. Central injections of alpha-methyl-p-tyrosine (80 micrograms/insect) reduced dopamine concentrations in the cerebral ganglia. However, neither 6-hydroxydopamine (20 micrograms/insect) nor 5,7-dihydroxytryptamine (20 micrograms/insect) caused reductions in amine levels when applied near or directly into the cerebral ganglia. The results suggest that specific lesions of aminergic neurons in insects by either 6-hydroxydopamine or 5,7-dihydroxytryptamine are impractical. The specific, long-lasting depletion of 5-HT by alpha-methyltryptophan suggests that this chemical may be useful in elucidating the functions of 5-HT in insects.     

Sympathetic and hyperthermic reactions by orexin A: role of cerebral catecholaminergic neurons

This experiment tested the effect of a lesion of cerebral catecholaminergic neurons on the sympathetic and thermogenic effects induced by an intracerebroventicular (icv) injection of orexin A. The firing rates of the sympathetic nerves to the interscapular brown adipose tissue (IBAT), along with IBAT, colonic temperatures and heart rate were monitored in urethane-anesthetized male Sprague-Dawley rats before an injection of orexin A (1.5 nmol) into the lateral cerebral ventricle and over a period of 150 min after the injection. Three days before the experiment, the rats were pre-treated with an icv injection of 6-hydroxydopamine (6-OHDA) or 6-OHDA plus desipramine or saline. The results show that orexin A increases the sympathetic firing rate, IBAT, colonic temperatures and heart rate in the rats pre-treated with saline. This increase is blocked by the pre-treatment with 6-OHDA alone or 6-OHDA plus desipramine. These findings indicate that cerebral catecholaminergic neurons (particularly the dopaminergic pathway) play a fundamental role in the complex reactions related to activation of the orexinergic system.     

Calcitonin inhibition of physiological and stimulated prolactin secretion in rats

The administration of salmon Calcitonin (sCT) intravenously (2.5 or 10 μg/kg) or into the lateral cerebral ventricles (2.5 or 25 ng/rat, i.c.v.) of unanaestized male rats induced clearcut decreases in plasma prolactin(PRL) levels. The i.c.v. injection of one of these doses of sCT (25 ng/rat) into rats with median eminence lesions was completely ineffective, while it induced a dramatic decrease in plasma PRL levels of sham-operated rats. Morphine- and heat stress-stimulated PRL levels were also abolished by sCT injection (250 ng/rat i.c.v.). The sCT-induced decrease in PRL levels was completely overcome by haloperidol, a dopamine-receptor blocker. We conclude that sCT may affect PRL secretion via an hypothalamic system, probably involving dopaminergic neurons. The present results indicate that CT, like many others peptides, may affect PRL secretion, directly or indirectly, even though further research is necessary to determine whether this effect has pharmacological or physiological importance.     

Retrogression of the metabolic activity of astrocytes after extinction of an experimental epileptogenic focus. Histochemical investigation

Previous anatomochemical and experimental investigations had resulted in the histochemical characterisation of the epileptogenic focus by the presence of "activated astrocytes"; cortical reactive astrocytes acquiring an intense activity of sundry dehydrogenases (DH) and becoming visible in spite of the positivity of the neuropil, and stopping at the amylose step in the synthesis of glycogen. This activation exists from the first electrical sings of epilepsy and even preceeds them in the case of glucose-6-phosphate DH. The present study deals with the fate of activated astrocytes after the disappearance of electrical signs of the epileptogenic focus produced by implantation of a cobalt powder-gelatin pellet into the cerebral cortex of the rat. After extinction of the focus, high activity of DH persists in astrocytes only in the immediate vicinity of the glial scar surrounding the implant. Later, among the DH investigated only glutamate and specially glucose-6-phosphate DH maintain an activity intense enough to reveal these cells on the background of the neuropil. These results point to a progressive return to a normal metabolism of astrocytes after the disappearance of the electrical signs of epilepsy by way of steps similar to those preceeding these signs.     

Effect of actinomycin D on the ultrastructure of the hippocampal cortex

Ultrastructure of the CA1 zone in the rat dorsal hippocampus has been investigated after injection of actinomycin D into the cerebral lateral ventricles. Actinomycin D possesses a wider spectrum of action as it was previously thought. The data obtained make it possible to suppose that certain cerebral disturbances (in particular, memory), produced with actinomycin D, can be dependent on or stipulated by: decreased DNA synthesis in the neuronal nuclei, disorder of RNA synthesis in neurons and astrocytes, damage of the protein synthesis apparatus only in neurons with a dense granular endoplasmic reticulum, decreasing contents of functionally active neurons, possessing a loose granular endoplasmic reticulum, decreasing production of energy by mitochondria of synapses, neurons and astrocytes.     

Intranuclear membranous inclusions in the CNS neurons of rats detectable after the administration of ascorbic acid and 6-hydroxydopamine

Intranuclear membranous inclusions were found in neurons of the rat's cerebral cortex and caudate nucleus 2-21 days following the intraperitoneal injection of ascorbic acid (0.2 and 2.0 g/kg) or the intracisternal infusion of 6-hydroxydopamine (300 micrograms). The intranuclear inclusions were mostly round, occasionally irregular in shape, consisting of one or several concentric membranes; in addition, they were electron-lucid with the diameter of 0.2-0.5 microns, sometimes up to 1 micron. Possible relationship between the formation of intranuclear membranous inclusions and the acceleration of intranuclear metabolism, particularly lipid peroxidation processes, are discussed.     

Total protein content and concentration in the neurons of the "mirror" epileptiform focus in the rat brain at the late stages of its existence]

The experimental lesion in rat brain was produced by the implantation of cobaltogelatin rod into the right parietalis anterior area. A quantitative measurement of total protein was made by microdensitometer in neurons controlateral to cobalt lesion at 112-115 days after cobalt implantation. In the experimental animals, protein content decreased in neurons of lamina III and V area parietalis anterior and the nucleus lateralis thalami by 6, 60 and 53%, resp. It has been concluded that different type neurons have different protein changes in response to paroxyzmal activity at late stages of epileptogenic mirror focus.     

Cholinotoxicity induced by ethylcholine aziridinium ion after intracarotid and intracerebroventricular administration

The effect of ethylcholine aziridinium ion (AF64A) after an intracerebroventricular (icv) injection was compared to that obtained after an intravascular administration. Reductions in choline acetyltransferase (ChAT) and acetylcholinesterase activities in the hippocampus but not in the cerebral cortex or the corpus striatum were observed 10 days after bilateral injection of AF64A into the rat cerebroventricles (3 nmol/side). However, when AF64A was injected into the carotid artery (1 mumol/kg) following a unilateral opening of the blood-brain barrier by a hypertonic treatment, a significant decrease in ChAT activity was observed in the ipsilateral side of the cerebral cortex but not in hippocampus, corpus striatum, or cerebellum. High-affinity choline transport was reduced significantly 11 days after an icv injection of AF64A in all the above mentioned brain regions, and recovered 60 days post injection in the cerebral cortex and in the corpus striatum but not in the hippocampus. Our results suggest that in various brain regions, AF64A causes various degrees of damage to cholinergic neurons, depending on the quantity of the toxin that reaches the target tissue.     

Phosphorylase and glucosyltransferases at the level of reactive astrocytes. A histochemical study]

Implantation of cobalt powder in the cerebral cortex of rat determines an epileptogenic focus where two types of reactive astrocytes are observed. The first type is mostly represented in the subcortical white matter but it does exist in the cortex around the implant. Phosphorylase and branching enzyme are both very active in these cells which are filled with glycogen. The second type is limited to the cortex and phosphorylase activity leads to an unbranched polysaccharid. These cells correspond to the "activated astrocytes" described by the authors in a previous paper and observed round irritative lesions which, in the cerebral cortex, produce epileptogenic foci.     

Ventricular injection of nerve growth factor increases dopamine content in the striata of MPTP-treated mice

Experimental depletion of dopaminergic striatal neurons was induced in mice with the neurotoxin MPTP. To investigate a possible effect of nerve growth factor on the damaged neurons, we injected 4 g into the right cerebral ventricle of mice three days after the last administration of MPTP. We found a significant increase of dopamine and homovanillic acid in the striatum of MPTP treated mice after NGF administration when compared with dopamine and HVA levels in MPTP-treated control mice (p<0.001). The increase of dopamine and homovanillic acid seems to be related to a partial restorative effect of NGF on the damaged dopaminergic cells, since ventricular administration of NGF to normal mice did not increase dopamine or homovanillic acid contents above the levels measured in untreated controls. It appears that administration of nerve growth factor prcduces a beneficial effect on damaged dopaminergic neurons; this effect could be due to stimulation of neuron sprouting from neurons that survived the toxic effect of MPTP. The increase of dopamine levels was seen 8 days after injection of nerve growth factor and was maintained at least until day 25, showing a lasting persistence of the restorative effect.     

Chronic morphine administration decreases 5-hydroxytryptamine and 2-hydroxyindoleacetic acid content in the brain of rats

To study the effects of chronic morphine treatment on cerebral 5-hydroxytryptamine (5HT) metabolism morphine was administered twice daily for 5 or 8 weeks to male Wistar rats. Control rats were treated with 0.9% NaCl solution for the same period. In rats treated chronically with morphine for 8 weeks the cerebral concentrations of 5HT and 5HIAA were reduced by 12--15% (P less than 0.05) at 26--28 h after the last morphine injection (50 mg/kg s.c.). No such decrease was found in the brain of rats treated with morphine for 5 weeks. A test dose of morphine (30 mg/kg s.c. 2h) increased the cerebral concentration and probenecid-induced accumulation of 5HIAA in the rats treated with morphine for 8 weeks almost as much as in the brain of the control rats. Naloxone (10 mg/kg s.c. 2h) did not cause clear changes in the cerebral 5HT or 5HIAA concentration. These experiments suggest that endogenous opioid mechanisms are concerned in the regulation of 5HT neurons and that prolonged morphine treatment weakens these mechanisms. This weakening of endogenous regulation of 5HT neurons, which, however, still respond to acute morphine administration, might be part of the mechanism of compulsive drug use in narcotic addiction. It is possible that these neurons in dependent individuals do not function optimally without exogenous morphine. A similar phenomenon--weakening of endogenous regulation combined with clear responsivity to exogenous opiates--occurs in the cerebral dopamine neurons of rats treated chronically with narcotic analgesics.     

Serotonin-like immunoreactivity in the ventral nerve cord of the Colorado potato beetle,Leptinotarsa decemlineata: Identification of five different neuron classes

Summary In an immunohistochemical study of the ventral nerve cord of L. decemlineata, five distinct neuron categories were distinguished: 1) Two paired segmental twin interneurons occur in each ganglion or neuromere; their axons distribute processes over almost the entire nerve cord and run to the cerebral ganglion complex. In contrast, other axons are distributed locally. 2) Four large frontal neurosecretory neurons occur in the suboesophageal ganglion (SOG), two of which have axons that run into the mandibular nerves to form a neurohemal plexus on the surface of cerebral nerves. 3) A pair of large caudal neurons occur in the terminal ganglion and innervate the hindgut. 4) Local miniature interneurons occur in the SOG. 5) Terminal neurons are present in the last abdominal ganglion. Segmental twin interneurons appear to be grouped into 3 functional units spanning several ganglia. Their axons run to specific projection areas, which separate the functional units, and which mark the externally visible separation of condensed ganglion complexes. A possible role of the most caudal functional unit might be the synaptic control of caudal neurons innervating the hindgut.     

α-Guanidinoglutaric Acid in Cobalt-Induced Epileptogenic Cerebral Cortex of Cats

: Guanidino compounds in the cobalt-induced epileptogenic cerebral cortex of cats were fluorometrically analysed by a JASCO G-520 guanidino compounds analyser, and an unknown high peak was observed in the chromatogram that was identical to the peak of authentic α-guanidinoglutaric acid. In another experiment, the substance was extracted from the cobalt focus tissue, converted into dimethylpyrimidyl derivative-butylester, and analysed by a GC/MS technique. The mass spectrum of the substance was identical to the dimethylpyrimidyl derivative of α-guanidinoglutaric acid butylester (M⁺= 365).     

Electroencephalographic study of the effect of neurotoxin DSP-4 in iron model of chronic focal epilepsy.

The effect of the noradrenergic neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) on electroencephalographic activity (EEG) was studied in the model of chronic focal epilepsy induced by intracortical injection of FeCl3 in the rat. EEG activity was recorded from the epileptogenic focus (ipsilateral and contralateral) in chronic experiments before and after DSP-4 treatment. In some experiments EEG activity was also simultaneously recorded from the cortical epileptogenic focus and locus coeruleus before and after DSP-4 treatment to study the effect of iron-induced seizure activity and of DSP-4 on the locus coeruleus electrical activity. The results showed that DSP-4 aggravated the iron-induced epileptiform activity as well as the locus-coeruleus electrical activity. The data also showed that, induction of epilepsy by FeCl3 is accompanied by enhancement of the locus coeruleus electrical activity. Our study demonstrates that DSP-4 intensifies and modifies the epileptic activity in the iron-induced chronic epilepsy model and that the effects of toxin persist for a longer duration.