Inhibitory effects of intravenous anaesthetic agents on K+-evoked glutamate release from rat cerebrocortical slices. Involvement of voltage-sensitive Ca2+ channels and GABA(A) receptors
ABSTRACT It is widely accepted that most general anaesthetic agents depress the central nervous system (CNS) by potentiation or activation of the GABA(A) receptor-mediated Cl(-) conductance. These agents also reportedly inhibit voltage-sensitive Ca(2+) channels (VSCCs), thus depressing excitatory transmission in the CNS. However, in this regard there are few functional data at the level of neurotransmitter release. In this study we examined the effects of VSCC antagonists and a range of intravenous anaesthetic agents on K(+)(40 mM)-evoked glutamate release from rat cerebrocortical slices in the absence and presence of the GABA(A) receptor antagonist bicuculline (100 microM). We employed both selective and non-selective VSCC antagonists, the anaesthetic barbiturates thiopental, pentobarbital and phenobarbital, the non-anaesthetic barbiturate barbituric acid, the non-barbiturate anaesthetics alphaxalone, propofol and ketamine and the GABA(A) receptor agonist, muscimol. Glutamate released into the incubation medium was determined by a glutamate dehydrogenase-coupled assay. Omega-agatoxin IV(A) (P-type VSCC), omega-conotoxin MVII(C) (P/Q-type VSCC) and Cd(2+) (non-selective) essentially abolished glutamate release whilst nifedipine (L-type VSCC) and omega-conotoxin GVI(A) (N-type VSCC) reduced release by less than 30%. The concentrations producing 50% of the maximum inhibition (IC(50)) for thiopental, pentobarbital, phenobarbital, alphaxalone, propofol and ketamine were (in microM) 8.3, 22, 112, 6.3, 83 and 120, respectively. Barbituric acid produced a small (about 20%) inhibition. With the exception of ketamine, the IC(50) values for these anaesthetic agents were increased threefold by bicuculline (100 microM). In addition, muscimol significantly inhibited release by 26% with an IC(50) of 1.1 microM. In summary, a range of anaesthetic agents at clinically achievable concentrations inhibit glutamate release and this inhibition of release appears to be due mainly to direct inhibition of P/Q-type VSCCs, although activation of the GABA(A) receptor plays a role in this response.
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ABSTRACT: Orexins (OXs) regulate sleep with possible interactions with brain noradrenergic neurons. In addition, noradrenergic activity affects barbiturate anesthesia. As we have also recently reported that OXs selectively evoke norepinephrine release from rat cerebrocortical slices we hypothesized that barbiturate anesthesia may result from of an interaction with central orexinergic systems. To test this hypothesis, we performed a series of in vivo and in vitro studies in rats. In vivo, the effects of i.c.v. OX A, B and SB-334867-A (OX1 receptor antagonist) on pentobarbital, thiopental or phenobarbital-induced anesthesia times (loss of righting reflex) was assessed. In vitro effects of barbiturates and SB-334867-A on OX-evoked norepinephrine release from cerebrocortical slice was examined. In Chinese hamster ovary cells expressing human OX1/OX2 receptors OX A- and B-evoked increases in intracellular Ca2+ were measured with and without barbiturates. OX A and B significantly decreased pentobarbital, thiopental and phenobarbital anesthesia times by 15-40%. SB-334867-A increased thiopental-induced anesthesia time by approximately by 40%, and reversed the decrease produced by OX A. In vitro, all anesthetic barbiturates inhibited OX-evoked norepinephrine release with clinically relevant IC50 values. A GABAA antagonist, bicuculline, did not modify the inhibitory effects of thiopental and the GABAA agonist, muscimol, did not inhibit norepinephrine release. In addition there was no interaction of barbiturates with either OX1 or OX2 receptors. Collectively our data suggest that orexinergic neurons may be an important target for barbiturates, and GABAA, OX1 and OX2 receptors may not be involved in this interaction.Neuroscience 02/2003; 121(4):855-63. DOI:10.1016/S0306-4522(03)00554-2 · 3.33 Impact Factor
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ABSTRACT: Understanding the impact of active dendritic properties on network activity in vivo has so far been restricted to studies in anesthetized animals. However, to date no study has been made to determine the direct effect of the anesthetics themselves on dendritic properties. Here, we investigated the effects of three types of anesthetics commonly used for animal experiments (urethane, pentobarbital and ketamine/xylazine). We investigated the generation of calcium spikes, the propagation of action potentials (APs) along the apical dendrite and the somatic firing properties in the presence of anesthetics in vitro using dual somatodendritic whole cell recordings. Calcium spikes were evoked with dendritic current injection and high-frequency trains of APs at the soma. Surprisingly, we found that the direct actions of anesthetics on calcium spikes were very different. Two anesthetics (urethane and pentobarbital) suppressed dendritic calcium spikes in vitro, whereas a mixture of ketamine and xylazine enhanced them. Propagation of spikes along the dendrite was not significantly affected by any of the anesthetics but there were various changes in somatic firing properties that were highly dependent on the anesthetic. Last, we examined the effects of anesthetics on calcium spike initiation and duration in vivo using high-frequency trains of APs generated at the cell body. We found the same anesthetic-dependent direct effects in addition to an overall reduction in dendritic excitability in anesthetized rats with all three anesthetics compared with the slice preparation.Journal of Neurophysiology 04/2008; 99(3):1394-407. DOI:10.1152/jn.01126.2007 · 3.04 Impact Factor
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ABSTRACT: This animal study tested the spatial learning memory of "depressed" rats undergoing electroconvulsive therapy (ECT) or ECT combined with propofol and aimed to reveal the glutamatergic mechanisms in the hippocampus. Sixty Sprague-Dawley rats were randomly divided into 5 groups (n = 12 rats per group): control group, depression group, propofol group, ECT group, and propofol + ECT group. Rats were stressed repeatedly for 21 days to establish depression model. After the model was set up, rats of the propofol group were administrated with propofol (100 mg/kg). Rats of ECT group were administered ECT once on alternate days for 2 weeks. ECT + propofol group rats were given ECT after anesthesia with propofol (100 mg/kg). Spatial memory was assessed by Morris water maze. Glutamate content in hippocampus was measured by chromatometry. N-methyl d-aspartate (NMDA)-NR2B expression was detected by immunohistochemistry. After treatment, the behavior level of rats in ECT group and ECT + propofol group was higher than that in depression group, and there was no significance between ECT group and ECT + propofol group. The evasive latency of rats detected by Morris water maze got shorter and shorter from the first day to fourth day. The evasive latency in ECT group was longer than that in ECT + propofol group and depression group, and the evasive latency in ECT + propofol group was shorter than that in depression group. Glutamate contents in hippocampus of rats in depression group and propofol group were higher than those in other groups, and glutamate content in ECT group was lower than that in other groups. The content in ECT + propofol group was lower than that in depression group, but higher than that in ECT group. N-methyl d-aspartate-NR2B expression in hippocampus of rats in depression group was lower than that in control group, but the expressions in ECT group and ECT + propofol group were higher than that in control group, and the expression in ECT + propofol group was lower than that in ECT group. The glutamate content in hippocampus of depressed rats heightens, and the NMDA-NR2B expression down-regulated, which may cause "depression" symptoms and learning memory impairment. After ECT, the glutamate contents decreased, and NMDA-NR2B expression up-regulated, the depression symptoms improved, and the spatial memory worsened simultaneously. However, propofol inhibited the excessive decrease of glutamate and excessive up-regulation of NMDA-NR2B caused by ECT, and both the depression symptoms and the spatial memory of depressed rats improved.The journal of ECT 06/2010; 26(2):126-30. DOI:10.1097/YCT.0b013e3181a9947a · 1.39 Impact Factor