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Abstract Background Many sedative agents, including anesthetics, produce explicit memory impairment by largely unknown mechanisms. Sharp-wave ripple (SPW-R) complexes are network activity thought to represent the neuronal substrate for information transfer from the hippocampal to neocortical circuits, contributing to the explicit memory consolidation. In this study we examined and compared the actions of two barbiturates with distinct amnesic actions, the general anesthetic thiopental and the anticonvulsant phenobarbital, on in vitro SPW-R activity. Results Using an in vitro model of SPW-R activity we found that thiopental (50–200 μM) significantly and concentration-dependently reduced the incidence of SPW-R events (it increased the inter-event period by 70–430 %). At the concentration of 25 μM, which clinically produces mild sedation and explicit memory impairment, thiopental significantly reduced the quantity of ripple oscillation (it reduced the number of ripples and the duration of ripple episodes by 20 ± 5%, n = 12, P < 0.01), and suppressed the rhythmicity of SPWs by 43 ± 15% (n = 6, P < 0.05). The drug disrupted the synchrony of SPWs within the CA1 region at 50 μM (by 19 ± 12%; n = 5, P < 0.05). Similar effects of thiopental were observed at higher concentrations. Thiopental did not affect the frequency of ripple oscillation at any of the concentrations tested (10–200 μM). Furthermore, the drug significantly prolonged single SPWs at concentrations ≥50 μM (it increased the half-width and the duration of SPWs by 35–90 %). Thiopental did not affect evoked excitatory synaptic potentials and its results on SPW-R complexes were also observed under blockade of NMDA receptors. Phenobarbital significantly accelerated SPWs at 50 and 100 μM whereas it reduced their rate at 200 and 400 μM. Furthermore, it significantly prolonged SPWs, reduced their synchrony and reduced the quantity of ripples only at the clinically very high concentration of 400 μM, reported to affect memory. Conclusion We hypothesize that thiopental, by interfering with SPW-R activity, through enhancement of the GABAA receptor-mediated transmission, affects memory processes which involve hippocampal circuit activation. The quantity but not the frequency of ripple oscillation was affected by the drug.

Abstract: Several studies have indicated a functional differentiation across the septotemporal axis of rat hippocampus. Our previous results have shown that the α1β2γ2-GABAA receptor subtype dominates in dorsal hippocampus (DH), while the α2β1γ2-subtype prevails in ventral hippocampus (VH). We therefore studied possible differences in the pharmacological properties and receptor binding parameters of the GABAA receptor subtypes between DH and VH, by examining: (1)(a) the specific binding of [3H]-flunitrazepam (Benzodiazepine sites agonist) by using quantitative autoradiography, (b) the kinetic parameters of [3H]-flunitrazepam specific binding, by using the “wipe off” technique and (2) the competitive displacement of [3H]-flunitrazepam binding by using zolpidem (selective agonist of the α1-subtype) and L-655,708 (selective inverse agonist of the α5-subtype) and the enhancement of [3H]-flunitrazepam binding by using etomidate (selective positive modulator of the β2-subunit), in an autoradiographical saturation kinetic study. Our results showed in VH compared to DH: (A) lower level of [3H]-flunitrazepam binding, apparently due to weaker binding affinity (higher K D value), since no differences in the B max value could be detected, (B) higher IC50 values for zolpidem and lower IC50 values for L-655,708 and (C) higher EC50 values for etomidate. In conclusion, the lower binding for zolpidem and etomidate and the higher binding for L-655,708 observed in VH support the evidence that the α1β2γ2-GABAA receptor subtype dominates in DH and the α5-subtype prevails in VH. Further, our results suggest differential pharmacological effects of the benzodiazepines in DH compared to VH, with the sedative effects being more potent in the dorsal hippocampus. [Copyright &y& Elsevier]

Abstract: The effects of the hexapeptide angiotensin II (3–8) ANG IV, the selective A1 receptor agonist cyclohexyladenosine (CHA) and the combination of ANG IV + CHA on pentylenetetrazol (PTZ)-generalized seizures; kindling development and maintenance were studied. By using in vitro quantitative receptor autoradiography, the regulation of adenosine A1 receptor density at different time points during the kindling procedure and postkindling period was determined. ANG IV and CHA effectively reduced clonic seizures in PTZ-generalized seizure model, in PTZ-kindled mice as well as during kindling development and a week later by rechallenge with PTZ. Furthermore, coadministration of ANG IV and CHA had a strong anticonvulsant effect, both compounds acting synergistically. A significant increase of adenosine A1 receptor density was detected in somatosensory cortex, hippocampus, amygdala and geniculate nuclei early in the kindling procedure (after the 3rd injection), which persisted at least 1 month after the end of kindling procedure. In addition, a delayed up-regulation of adenosine A1 receptor binding was observed a week after kindling in the mamillary bodies and a month later in the motor cortex. The pretreatment with ANG IV caused a down-regulation of adenosine A1 receptor density to the control level in most time points and brain areas. In conclusion, PTZ kindling-induced increase of adenosine A1 receptor binding at different time points and in specific brain structures might represent an adaptive mechanism for coping with the hyperexcitability typical for this phenomenon. The antiepileptogenic effect of ANG IV could be realized partly through an adenosine-dependent mechanism. [Copyright &y& Elsevier]

The present study examined the effect of pentylenetetrazol (PTZ) induced kindling as well as the action of the hexapeptide angiotensin IV (ANG IV) on the dopamine (DA) D1 and D2 receptor binding in the basal ganglia of the mouse brain. By using quantitative receptor autoradiography, it was found that PTZ kindling led to a decrease in DA D2 receptor density (about 20%) in all regions of the neostriatum (NS) as well as in the olfactory tubercle (OT), the nucleus accumbens (NA) and the globus pallidus, which persisted 24 h and 7 days after the kindling procedure. PTZ induced kindling also elicited a decrease in DA D1 receptor binding sites (about 10%), which however was, restricted to the rostral NS (rNA) and NA. ANG IV (0.2 mg/kg), injected prior to PTZ, not only prevented the development of the kindling process but it also reversed the kindling-induced down-regulation of both DA receptors to the control levels. Furthermore ANG IV induced an area-specific increase of DA D1 receptor density above control levels in the dorsal part of rNS. These findings suggest that DA D2 receptors could mainly contribute to epileptogenesis in the PTZ kindling model, whereas the role of DA D1 receptors is limited to particular regions in the basal ganglia. The anticonvulsant effect of ANG IV pretreatment might be influenced by a DA-related mechanism and particularly by preventing D2 receptor down-regulation as well as by an adaptive area-specific increase in DA D1 receptors. [Copyright &y& Elsevier]

Recent data demonstrate weaker gamma-aminobutyric acid (GABA)-ergic inhibition in ventral (VH) compared with dorsal (DH) hippocampus. Therefore, we examined possible differences regarding the GABAA receptors between VH and DH as follows: 1) the expression of the GABAA receptor subunits (alpha1/2/4/5, beta1/2/3, gamma2, delta) mRNA and protein and 2) the quantitative distribution and kinetic parameters of [3H] muscimol (GABAA receptor agonist) binding. VH compared with DH showed: 1) lower levels for alpha1, beta2, gamma2 but higher levels for alpha2 and beta1 subunits in CA1, CA2, and CA3, the differences being more pronounced in CA1 region; in the CA1 region, the mRNA levels of alpha5 were higher, whereas those of alpha4 subunit were slightly lower; in dentate gyrus, the mRNA levels of alpha4, beta3, and delta subunits were significantly lower, presumably suggesting a lower expression of the alpha4/beta3/delta receptor subtype; and 2) lower levels of [3H]muscimol binding, with the lowest value observed in CA1, apparently resulting from weaker binding affinity, insofar as the KD values were higher in VH, whereas the Bmax values were similar between DH and VH. The differences in the subunit expression and the lower affinity of GABAA receptor binding observed predominantly in the CA1 region of VH suggest that the alpha1/beta2/gamma2 GABAA receptor subtype dominates in DH, and the alpha2/beta1/gamma2 subtype prevails in VH. This could underlie the lower GABAA-mediated inhibition observed in VH and, to some extent, explain 1) the higher liability of VH for epileptic activity and 2) the differential involvement of DH and VH in cognitive and emotional processes.