Long-term reduction of benzodiazepine receptor density in the rat cerebellum by acute seizures and kindling and its recovery 6 months later by a pentylenetetrazole challenge.

Seizures induced by an acute pentylenetetrazole (50 mg/kg) injection were accompanied by a long-term (at 1-48 h, but not on day 7) decrease in the density (B(max)) of [3H]-diazepam binding to benzodiazepine receptors in rat cerebellar cortex with no change in affinity (K(d)). Kindling for 24 days by daily administrations of pentylenetetrazole (20 mg/kg) led to the same decrease in benzodiazepine receptor density (at 1-48 h, but not on day 7) as that observed after a single dose of pentylenetetrazole (50 mg/kg). This suggests a common mechanism for both acute and kindling-induced seizures, dependent on the long-term receptor changes. The increased susceptibility to seizures persisted for 6 months after the termination of kindling, with BDZ receptor density in cerebellar cortex reduced almost by half. In age-matched controls, an acute dose of PTZ (30 mg/kg) induced seizures and decrease in both B(max) and K(d) of [3H]-diazepam binding. In kindled rats, at 6 months post-kindling, the same dose of PTZ (30 mg/kg) restored the benzodiazepine receptor density to the level found 6 months before, at the time of termination of kindling. Also, the severity of seizures was enhanced in the kindled rats. The results are discussed in terms of a balance of inhibitory and excitatory processes, in which the reduced BDZ receptor density at 6 months post-kindling may represent a compensatory reaction to outbalance some alterations in excitatory systems that have been reported to be induced by kindling.

[The properties of the benzodiazepine receptors of the rat cerebellum 6 months after korazol-induced kindling and recurrent seizures]. / Svoistva benzodiazepinovykh retseptorov mozzhechka krys cherez shest' mesiatsev posle korazolovogo kindlinga i povtornykh sudorog.

It was shown that the increased brain seizure readiness persisted within 6 months after termination of corazol kindling. Seizures of the same severity as during kindling (corazol injection in a dose of 20 mg/kg) were reproduced by corazol injection in a dose of 30 mg/kg. In contrast to the control rats, in this situation an autoenhancement of seizures was observed in the kindled animals. Acute corazol seizures induced a decrease in Bmax and Kd of 3H-diazepam binding with benzodiazepine receptors (BDR) in the cerebellum of the 10-months-old control rats white the young animals demonstrated only a decrease in Bmax of binding. In 6 months after kindling termination the BDR activity (Bmax) was reduced by one half. However, we think that the increase in Bmax is not responsible for persistence of the increased seizure readiness. It seems possible that down regulation of receptor activity develops independently of kindling but in response to long-lasting corazol application. Probably, Bmax spontaneously decreases after the termination of the long-term corazol application. The single dose of corazol (30 mg/kg) restores the changes in BDR density to the level when seizure readiness has been just fixed (6 months after kindling termination), independently of the primary receptor density.

[The properties of the benzodiazepine receptors in the rat cerebellum after acute seizures and the development of korazol-induced kindling]. / Svoistva benzodiazepinovykh retseptorov mozzhechka krys posle ostrykh sudorog i razvitiia korazolovogo kindlinga.

The acute korazol (pentylenetetrazol) injection (50 mg/kg) induced seizures which were accompanied by a long-lasting (from 30 minutes to 3 days) decrease in benzodiazepine receptor (BDR) density (Bmax) in rat cerebellum without change in affinity. The density of the BDR was normalized on the 7th day after seizure termination. There were no differences in the initial BDR characteristics between the animals more sensitive to korazol (a dose of 25 mg/kg was sufficient for seizure induction) and less sensitive (30 mg/kg were ineffective). The chronic daily (for 24 days) administration of korazol in a subconvulsive dose led to an increase in seizure readiness (kindling). In 30 min after the last korazol injection the BDR density was decreased to the same extent as after the acute 50 mg/kg korasol administration. The BDR density was normalized on the 7th day after kindling. It was demonstrated that the high-dose-induced and after-kindling seizures were underlain by the same mechanisms. The results suggest that the development of kindling depends on the state of the long-lasting receptors rather than the development of kindling forms the long-lasting reactions. The process of summation is at the basis of kindling development. The long-lasting decrease in activity of BDR receptors induced by a subconvulsive dose of korazol is summed with the following effect of the same dose.

[The action of corticosterone on the macroerg pool and membrane permeability in rat hippocampal slices during hypoxia]. / Deistvie kortikosterona na pul makroérgov i sostoianie membrannoi pronitsaemosti v srezakh gippokampa krys pri gipoksii.

Incubation of rat hippocampal slices for 30 min in medium containing 900, 700, or 500 microM of O2 did not significantly change ATP, ADP, or CP levels, but when O2 content dropped to 100-200 microM the levels of ATP and CP decreased by 2 to 5 times. Under the same conditions CPK maximal solubilization was shown. Corticosterone acetate (CSA, 160 microM) did not show a membranotropic effect, nor did it influence the macroerg pool under conditions of high O2 content, but when O2 concentration was 100 microM a twofold increase of ATP and CP levels was observed. This effect correlated with a twofold increase of CPK membrane permeability. The results of this study permit us consider that membrane labilization is not a reflection of CSA neurotoxic effect in hypoxia. A relationship between CSA antihypoxic effect and macroerg pool increase is discussed.

[Changes in the level of cyclic nucleotides in the left and right sensorimotor areas of the cerebral cortex in rats after corazol-induced kindling]. / Izmeneniia urovenia tsiklicheskikh nukleotidov v levoi i pravoi sensomotornoi oblasti kory mozga krys posle korazolovogo kindlinga.

Levels of cAMP and cGMP in the left and right areas of the rat sensorimotor cortex (SMC) 7 days after pentylenetetrazole kindling (30 days) were determined by radioimmunological assay. It is shown that levels of cyclic nucleotides remained unchanged in rats SMC 7 days after single injection of seizure dose (70 mg/kg) of pentylenetetrazole. Levels of cAMP and cGMP in SMC increased by 2-3 times after 30-day administration of physiological saline as compared to single injection; cortical asymmetry of cAMP and cGMP is absent. Differences in the levels of cyclic nucleotides in sensitive and low-sensitive animals to pentylenetetrazole were not revealed. When kindling was completed cGMP level increased in right SMC and it decreased in left SMC as compared to control animals; cAMP level was unaltered in left and increased in right SMC. The role of cortical asymmetry of cyclic nucleotides development of seizure state is discussed.

[Differences in the hormonal status of rats highly and poorly resistant to hypoxia]. / Razlichiia gormonal'nogo statusa u vysoko- i nizkoustoichivykh k gipoksii krys.

Rats were tested for hypobaric hypoxia and decapitated two weeks later at 9-12, 13-14 and 15-17 o'clock. The levels of corticosterone, insulin and testosterone were determined in serum by RIA. Great daily fluctuations and higher elevation of corticosterone level in response to additional 1.5-min hypoxia were observed in low hypoxia-resistant (HHR) rats. The correlation of corticosterone/insulin ratio was directly proportional in LHR rats and inversely proportional in HHR rats. No difference in testosterone and insulin levels in LHR and HHR rats was found.

[Organic calcium antagonists verapamil and ryodipine prevent an increase of free calcium in synaptosomes of the rat brain during corazol kindling]. / Organicheskie antagonisty kal'tsiia verapamil i riodipin predotvrashchaiut uvelichenie svobodnogo kal'tsiia v sinaptosomakh mozga krys v techenie korazolovogo kindlinga.

In experiments on Wistar male rats it was shown that i.p. administration of verapamil or ryodipine (1,4-dihydropyridine) 15 min before each daily injection of pentylenetetrazol (PTZ) in a subconvulsive dose 30 mg/kg during 28 days significantly delayed the development of PTZ-induced kindling and attenuated kindled seizure reactions during 21-23 days as compared with control. One week after kindling an increase in free Ca2+ concentration in control rat brain synaptosomes by 60% was revealed; the treatment by verapamil and ryodipine prevented this increase. Nevertheless, Ca2+ antagonists studied at least under our experimental conditions inspite of a significant decrease in free Ca2+ concentration in synaptosomes would not completely prevent the development of kindling. Thus, an alteration in calcium homeostasis will not be the only mechanism of plasticity and long-term changes of neurons during kindling.

[Effects of corazol on the regulation of GABAa receptor-channel complex]. / Deistvie korazola na reguliatsiiu GAMKa-kanal-retseptornogo kompleksa.

Effects of corazol (pentylenetetrazole, PTZ) on the GABAA receptor-channel complex mediated 36Cl- influx into synaptoneurosomes isolated from rat cerebral cortex, were studied. PTZ inhibited the 36Cl- influx stimulated by a GABA agonist, muscimol, concentration-dependently with an IC50 of 2.11 +/- 0.25 mM. In addition, the rate of muscimol-dependent desensitization of the GABAA receptor-channel complex was significantly reduced in the presence of PTZ. Apparently, the superimposition of these two mutually opposing effects is the cause of a mixed (competitive/noncompetitive) type of inhibition displayed by PTZ. The rate of desensitization of GABAA receptor-channel complex is suggested to be regulated by intracellular concentration of Cl- through a feed-back mechanism.

[Effects of magnesium and nickel ions on penicillin-induced focal epileptic activity in the cerebral cortex of rats]. / Vliianie ionov magniia i nikelia na fokal'nuiu penitsillin-indutsirovannuiu épilepticheskuiu aktivnost' v kore golovnogo mozga krys.

In experiments on freely moving male Wistar rats on the model of penicillin-induced focal epileptic activity (EA) (the application onto the sensorimotor cortex of a filter paper soaked with benzylpenicillin sodium salt solution) it was shown that addition of MgSO4 (series 1) and NiCl2 (series 2) into the solution of penicillin significantly weakened EA. The combination of Mg2+ and Ni2+ with penicillin (series 3) produced a more significant suppression of EA as compared with separate application of the above-mentioned ions: the latency period of appearance of interictal discharges (IID) increased, the frequency and amplitude of IID decreased much more, no ictal discharges appeared in any animal, the duration of epileptic foci reduced to a much greater extent. This effect can be explained by the blockade of Ca current by the above-mentioned ions. One can suppose that the amplification of antiepileptic effects of combined action of Mg2+ and Ni2+ was due to an increase in the number of blocked voltage-dependent and NMDA-operated calcium channels.

[Anti-epileptic effect of the new calcium channel blocker IOS-1.1212]. / Antiépilepticheskie éffekty novogo blokatora Ca-kanalov IOS-1.1212.

In experiments on freely moving male Wistar rats it was shown that IOS-1.1212 (1,4-dihydropyridine) in a dose 2 and 10 mg/kg (i. p.) suppressed the penicillin-induced focal epileptic activity in cerebral cortex. Similar suppressing effect of IOS-1.1212 was shown on acute generalized tonic-clonic pentylenetetrazol (PTZ) seizures (75 mg/kg i. p.) and on chronic PTZ administration (PTZ-kindling, 30 mg/kg i. p. during 30 days): when injected 30 min before each PTZ administration it delayed the development of kindling-induced seizures susceptibility in randomized animals (series 1) and attenuated the severity of seizures in PTZ-sensitive animals (series 2). However, IOS-1.1212 had no effect on the strychnine-induced focal epileptic activity. In male Icr:Icl mice IOS-1.1212 in a dose 1.5 and 5 mg/kg also influenced the PTZ convulsions (i. v. titration of 1% solution at a rate of 0.01 ml/s) and had no effect on the strychnine convulsions (i. v. titration of 0.01% solution at a rate of 0.01 ml/s) and on maximal electroshock. In addition, IOS-1.1212 significantly increased antiepileptic effect of phenobarbital on maximal electroshock.

[Antiepileptic effects of nifedipine]. / Ob antiépilepticheskikh éffektakh nifedipina.

In experiments on freely moving male Wistar rats it was shown that nifedipine in a dose 10 mg/kg (i.p.) suppressed the penicillin-induced focal epileptic activity in cerebral cortex. A similar suppressing effect of nifedipine was shown on acute generalized tonic-clonic pentylenetetrazol (PTZ) seizures (75 mg/kg, i.p.). Nifedipine in the same dose was not effective on chronic PTZ administration (PTZ-kindling, 30 mg/kg i.p. during 28 days): when injected 30 min before each PTZ administration it didn't delay the development of kindling induced seizure susceptibility and had no effect on the severity of seizures. The administration of nifedipine in a dose of 10 or 30 mg/kg to control kindled animals which had not been treated with nifedipine had no influence on the severity of seizures provoked by a testing dose of PTZ (30 mg/kg i.p.): its intensity was similar to that of caused by PTZ injection along.

[Effect of a Ca-channel blocker, ryodipine, on focal and generalized epileptic activity]. / Vliianie blokatora Ca-kanalov riodipina na ochagovuiu i generalizovannuiu épilepticheskuiu aktivnost'.

The experiments were performed on 102 freely moving Wistar rats. Epileptic foci were produced by the application of a filter paper soaked in a sodium benzylpenicillin solution (20,000 IU/ml) onto sensorimotor cortex. It was shown that an intraperitoneal administration of ryodipine (1,2 and 5 mg/kg) during a steady epileptic activity (EA) resulted in suppression of EA in most animals. Antiepileptic effect of ryodipine was manifested by a decreased frequency and amplitude of interictal discharges and a less frequent appearance of ictal discharges (ID). Prior administration of ryodipine (2 mg/kg) 30 min before producing the focus of EA resulted in an increased latency and decreased number of ID, and shortening of the duration of the focus of EA. Generalized convulsions were induced by intraperitoneal of pentylenetetrazol (60 mg/kg). Ryodipine (2 mg/kg, 30 min before pentylenetetrazol) increased latency to first convulsive episodes and delayed the development of generalized tonic-clonic seizures.

[Inhibition by local anesthetics and barbiturates of the active transport of H+ in the synaptic vesicle membranes of the brain]. / Tormozhenie mestnymi anestetikami i barbituratami aktivnogo transportaH+ v membranakh sinapticheskikh puzyr'kov mozga.

The effects of local anesthetics and barbiturates on the ATP-dependent H+ transport in synaptic vesicle membranes from rat brain were studied using a fluorescent probe, acridine orange. Local anesthetics depressed the active H+ transport with the following order of potencies: tetracaine trimecaine lidocaine procaine. Respective IC50 values were 0.07, 0.28, 0.46 and 0.60 mM. The local anesthetics also disrupted the endogenous pH gradient seen in the absence of ATP. Barbiturates inhibited the active H+ transport showing IC50 values in the range of 2-5 mM except for benzobarbital and barbital characterized by IC50 values of 0.5 and 20 mM, respectively. The order of potencies was benzobarbital hexobarbital amobarbital pentobarbital phenobarbital barbital. The endogenous pH gradient was not affected by the barbiturates. The results show that local anesthetics disrupt the H+ transport by acting as permeable weak bases (uncouplers) whereas barbiturates are likely to block and anion channel which maintains electroneutrality of the H+ transport in the membrane of synaptic vesicles.

[Ca2+- and inositol-1,4,5-triphosphate induced release of Ca2+ in the microsomal fraction of the rat brain]. / Ca2+- i inozitol-1,4,5-trifosfat-indutsirovannoe osvobozhdenie Ca2+ v mikrosomnoi fraktsii mozga krys.

Using the fluorescent probes, Quin 2 and chlortetracycline, a comparative study of the Ca2+ and inositol-1.4.5-triphosphate (IP3)-induced Ca2+ release from rabbit skeletal muscle sarcoplasmic reticulum (SR) terminal cisterns and rat brain microsomal vesicles was carried out. It was shown that Ca2+ release from rat brain microsomal vesicles is induced both by IP3 and Ca2+, whereas that in SR terminal cisterns is induced only by Ca2+. Data from chlorotetracycline fluorescence analysis revealed that CaCl2 (50 microM) causes the release of 15-20% and 40-50% of the total Ca2+ pool accumulated in rat brain microsomal vesicles and rabbit SR terminal cisterns, respectively. Using Quin 2, it was found that IP3 used at the optimal concentration (1.5 mM) caused the release of 0.4-0.6 nmol of Ca2+ per mg microsomal protein, which makes up to 10-15% of the total Ca2+ pool. IP3 does not induce Ca2+ release in SR. Preliminary release of Ca2+ from brain microsomes induced by IP3 diminishes the liberation of this cation induced by Ca2+. It is suggested that brain microsomes contain a Ca2+ pool which is exhausted under the action of the both effectors, Ca2+ and IP3.

[Effect of the Ca ionophore A-23187 on the plasmatic and mitochondrial potentials of the brain synaptosomes in rats: fluorescence measurements]. / Vliianie Ca-ionofora A23187 na plazmaticheskii i mitokhondrial'nyi potentsialy sinaptosom mozga krys: fliuorestsentnye izmereniia.

The applicability of the potential-sensitive dye diS-C3-(5) for the study of A23187 + Ca2+ induced plasma membrane hyperpolarization was tested in rat brain synaptosomes. An appropriate dye synaptosome ratio was chosen for the fluorescence titration dye in Ca-free Krebs-Ringer solution. The fluorescence intensity of the probe was increased upon the addition of Ca2+ (1 microM) to the synaptosomes in the presence of A23187 (1 microM). The effect of Ca2+ + A23187 persisted in a Na+-free medium or when Na+ channels were inhibited by tetrodotoxin as well as in high K+-depolarized synaptosomes (75 microM KCl). In the presence of oligomycin or a protonophore (1 microM) the effect of Ca2+ + A23187 was suppressed. This suggests that the A23187-induced fluorescence increase is due to a depolarization of intrasynaptosomal mitochondria. Therefore, the use of the dye diS-C3-(5) for the study of Ca-induced hyperpolarization does not seem to be feasible unless a quantitative model of changes in fluorescence related to the plasma and mitochondrial membrane potentials is elaborated.