Search PubMed⌕ Search

Biomedical subjects

G R Siggins

Publications and source records attributed to G R Siggins.

At least 37 records · Page 2Linked to original sources
PubMed1997

Transgenic models to assess the pathogenic actions of cytokines in the central nervous system.

In order to better understand the actions of proinflammatory cytokines in the mammalian CNS, a transgenic approach was employed in which the expression of IL-6, IL-3 or TNF-alpha was targeted to astrocytes in the intact CNS of mice. Transgenic mice exhibited distinct chronic-progressive neurological disorders with neurodegeneration and cognitive decline due to IL-6 expression, macrophage/microglial-mediated primary demyelination with motor impairment due to IL-3 expression and lymphocytic meningoencephalomyelitis with paralysis induced by TNF-alpha expression. Thus, expression of specific cytokines alone in the intact CNS results in unique neuropathological alterations and functional impairments, thereby directly implicating these mediators in the pathogenesis of CNS disease.

Animals↗
PubMed1997

Somatostatin depresses excitatory but not inhibitory neurotransmission in rat CA1 hippocampus.

In rat CA1 hippocampal pyramidal neurons (HPNs), somatostatin (SST) has inhibitory postsynaptic actions, including hyperpolarization of the membrane at rest and augmentation of the K+ M-current. However, the effects of SST on synaptic transmission in this brain region have not been well-characterized. Therefore we used intracellular voltage-clamp recordings in rat hippocampal slices to assess the effects of SST on pharmacologically isolated synaptic currents in HPNs. SST depressed both (R, S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainate and N-methyl--aspartate (NMDA) receptor-mediated excitatory postsynaptic currents (EPSCs) in a reversible manner, with an apparent IC50 of 22 nM and a maximal effect at 100 nM. In contrast, SST at concentrations up to 5 microM had no direct effects on either gamma-aminobutyric acid-A (GABAA) or GABAB receptor-mediated inhibitory postsynaptic currents (IPSCs). The depression of EPSCs by SST was especially robust during hyperexcited states when polysynaptic EPSCs were present, suggesting that this peptide could play a compensatory role during seizurelike activity. SST effects were greatly attenuated by the alkylating agent N-ethylmaleimide, thus implicating a transduction mechanism involving the Gi/Go family of G-proteins. Use of 2 M Cs+ in the recording electrode blocked the postsynaptic modulation of K+ currents by SST, but did not alter the effects of SST on EPSCs, indicating that postsynaptic K+ currents are not involved in this action of SST. However, 2 mM external Ba2+ blocked the effect of SST on EPSCs, suggesting that presynaptic K+ channels or other presynaptic mechanisms may be involved. These findings and previous results from our laboratory show that SST has multiple inhibitory effects in hippocampus.

Animals↗
PubMed1997

Metabotropic glutamate receptors regulate N-methyl-D-aspartate-mediated synaptic transmission in nucleus accumbens.

We recorded intracellularly from core nucleus accumbens (NAcc) neurons in brain slices to study the regulation by metabotropic glutamate receptors (mGluRs) of pharmacologically isolated N-methyl--aspartate-mediated excitatory postsynaptic currents (NMDA-EPSCs). Monosynaptic NMDA-EPSCs, evoked by local stimulation, were isolated by superfusion of the non-NMDA and gamma-aminobutyric acid-A (GABAA) receptor antagonists, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 10 microM) and bicuculline (15 microM), respectively. Trans-1-aminocyclopentane-1,3-decarboxylic acid (trans-ACPD; 50 microM), a nonspecific group 1 and 2 mGluR agonist, had no effect on resting membrane potential (RMP) or input resistance of NAcc neurons. However, it consistently decreased NMDA-EPSC areas (time integrals) dose dependently (1-100 microM; EC50 = 8 microM) and reversibly. The specific group 1 mGluR agonists quisqualate (1-4 microM) and (RS)-3, 5-dihydroxyphenylglycine (DHPG; 100 microM) did not mimic the trans-ACPD effect on NMDA-EPSCs, nor did exposure of the slice to the group 1 mGluR antagonist (+)-2-amino-3-phosphonopropionic acid (-AP3, 0.4 mM) inhibit the trans-ACPD effect. The putative mGluR1 and mGluR2 antagonist (+)-alpha-methyl-4-carboxyphenylglycine (MCPG) at 0.5 mM failed to antagonize trans-ACPD effects but at 1 mM blocked them. Both the group 2 mGluR agonist (2S,3S, 4S)-alpha-(carboxycyclopropyl)-glycine (-CCG-I, 2 microM) and the group 3 mGluR specific agonist (+)-2-amino-4-phosphonobutyric acid (-AP4, 20 microM) attenuated NMDA-EPSC areas; the effect of -AP4 was blocked by the group 3 antagonist (S)-2-amino-2-methyl-4-phosphonobutanoic acid (MAP4; 0.5 mM). Exogenously applied NMDA, in the presence of tetrodotoxin to prevent presynaptic effects, induced inward currents that were decreased by 20 microM -AP4 but not by 10 microM trans-ACPD. These findings suggest that NMDA receptor-mediated neurotransmission in NAcc is under dual inhibitory regulation by group 2 and 3 metabotropic receptor subtypes: -AP4-sensitive receptors located postsynaptically and those sensitive to trans-ACPD located presynaptically.

Animals↗
PubMed1997

Naloxone blocks long-term depression of excitatory transmission in rat CA1 hippocampus in vitro.

In rat hippocampal slices, high intensity tetanic stimulation (two 1 s trains of 100 Hz separated by 20 s, 3-5X intensity of the test stimulus) of the Schaffer collateral-commissural (SCC) fibers induced a long-term depression (LTD) of the negative field excitatory postsynaptic potentials (fEPSP) in stratum radiatum of the CA1 region. The initial slope of the fEPSP, evoked by a single test shock applied to the SCC fibers, was depressed for a period longer than 40 min following such high intensity tetanic stimulation to this fiber system. However, the same tetanic stimulation delivered at low (test) intensity induced long-term potentiation (LTP) of the fEPSPs. Thus, similar patterns of stimulation can induce either LTP or LTD, depending on whether low- or high-intensity tetanic stimuli are delivered. The LTD induced by high strength tetanic stimulation was clearly blocked by the opioid antagonist naloxone (1 microM); however, the N-methyl-D-aspartate (NMDA) receptor antagonist D-2-amino-5-phosphonopentanoate (AP5; 50 microM) had no effect on the LTD. Our data suggest that the strong stimulation used for LTD induction may have activated other afferent fiber systems and/or local interneurons in addition to SCC fibers, such as the enkephalin-containing terminals of the perforant path (PP) projecting to the stratum lacunosum moleculare or opioid peptide-containing interneurons. Thus, the resulting release of endogenous opioid peptides could play a role in the cellular mechanisms involved in some forms of long-term synaptic depression.

2-Amino-5-phosphonovalerate↗
PubMed1996

A cortical neuropeptide with neuronal depressant and sleep-modulating properties.

Acetylcholine (ACh) plays a key role in the transitions between the different phases of sleep: Slow-wave sleep requires low ACh concentrations in the brain, whereas rapid-eye-movement (REM) sleep is associated with high levels of ACh. Also, these phases of sleep are differentially sensitive to a number of endogenous neuropeptides and cytokines, including somatostatin, which has been shown to increase REM sleep without significantly affecting other phases. Here we report the cloning and initial characterization of cortistatin, a neuropeptide that exhibits strong structural similarity to somatostatin, although it is the product of a different gene. Administration of cortistatin depresses neuronal electrical activity but, unlike somatostatin, induces low-frequency waves in the cerebral cortex and antagonizes the effects of acetylcholine on hippocampal and cortical measures of excitability. This suggests a mechanism for cortical synchronization related to sleep.

Acetylcholine↗
PubMed1996

Low ethanol concentrations enhance GABAergic inhibitory postsynaptic potentials in hippocampal pyramidal neurons only after block of GABAB receptors.

Despite considerable evidence that ethanol can enhance chloride flux through the gamma-aminobutyric acid type A (GABA/A/) receptor-channel complex in several central neuron types, the effect of ethanol on hippocampal GABAergic systems is still controversial. Therefore, we have reevaluated this interaction in hippocampal pyramidal neurons subjected to local monosynaptic activation combined with pharmacological isolation of the various components of excitatory and inhibitory synaptic potentials, using intracellular current- and voltage-clamp recording methods in the hippocampal slice. In accord with our previous findings, we found that ethanol had little effect on compound inhibitory postsynaptic potentials/currents (IPSP/Cs) containing both GABA/A/ and GABA/B/ components. However, after selective pharmacological blockade of the GABA/B/ component of the IPSP (GABA/B/-IPSP/C) by CGP-35348, low concentrations of ethanol (22-66 mM) markedly enhanced the peak amplitude, and especially the area, of the GABA/A/ component (GABA/A/-IPSP/C) in most CA1 pyramidal neurons. Ethanol had no significant effect on the peak amplitude or area of the pharmacologically isolated GABA/B/-inhibitory postsynaptic current (IPSC). These results provide new data showing that activation of GABAB receptors can obscure ethanol enhancement of GABA/A/ receptor function in hippocampus and suggest that similar methods of pharmacological isolation might be applied to other brain regions showing negative or mixed ethanol-GABA interactions.

Animals↗
PubMed1996

Arachidonate 5-lipoxygenase and its activating protein: prominent hippocampal expression and role in somatostatin signaling.

5-Lipoxygenase-activating protein (FLAP) is an 18-kDa integral membrane protein required, in peripheral cells, for the activation of 5-lipoxygenase (5-LO) and for the resulting synthesis of leukotrienes from arachidonic acid. In the brain, the leukotrienes have been implicated in several pathophysiological events and in the electrophysiological effect of somatostatin, yet the cellular origin and role of these messenger molecules are still poorly understood. In the present study, we used reverse transcriptase-polymerase chain reaction, in situ hybridization, and immunohistochemistry to demonstrate that 5-LO and FLAP are expressed in various regions of the rat brain, including hippocampus, cerebellum, primary olfactory cortex, superficial neocortex, thalamus, hypothalamus, and brainstem. Highest levels of expression were observed in cerebellum and hippocampus. In the latter we demonstrate the colocalization of 5-LO and FLAP in CA1 pyramidal neurons. Moreover, electrophysiological experiments show that selective inhibition of FLAP with the compound MK-886 (0.25-1 microM) prevents the somatostatin-induced augmentation of the hippocampal K+ M-current. Our results provide necessary evidence for the presence and signaling role of 5-LO and FLAP in central neurons and strongly support their proposed participation in somatostatin-receptor transmembrane signaling.

5-Lipoxygenase-Activating Proteins↗
PubMed1996

Acamprosate (calcium acetylhomotaurinate) enhances the N-methyl-D-aspartate component of excitatory neurotransmission in rat hippocampal CA1 neurons in vitro.

The taurinate analog acamprosate (calcium acetylhomotaurinate) has received considerable attention in Europe for its ability to prevent relapse in abstained alcoholics. To determine the mechanism of acamprosate actions in the CNS, we superfused acamprosate onto rat hippocampal CA1 pyramidal neurons using an in vitro slice preparation. In current-and voltage-clamp recordings, acamprosate (100 to 100 microM) superfusion had little effect on resting membrane potential or input slope resistance. Acamprosate had no effect on Ca(2+)-dependent action potentials when tetrodotoxin was used to block Na+ spikes. In whole-cell voltage-clamp recordings, and in the presence of tetraethylammonium and Cs+ to block K+ channels, acamprosate had little effect on a Cd(2+)-sensitive inward current likely to be a high voltage-activated Ca2+ current. However, in both current- and voltage-clamp recordings, acamprosate significantly increased the N-methyl-D-aspartate (NMDA) component of excitatory postsynaptic potentials evoked by stimulation of Schaffer collaterals in the stratum radiatum, in the presence of the selective non-NMDA (R,S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-proprionic acid kainate) glutamate receptor antagonist 6-cyano-7-nitro-quinoxaline-2,3-dione and the GABAA receptor antagonist bicuculline. Acamprosate had inconsistent or no effects on the stratum radiatum-evoked non-NMDA component of the excitatory postsynaptic potentials, in the presence of bicuculline and the NMDA antagonist DL-2-amino-5-phosphonovalerate. Acamprosate, on average, had little effect on the late inhibitory postsynaptic potentials thought to be mediated by GABAB receptors. In the presence of tetrodotoxin to block synaptic transmission, acamprosate dramatically increased inward current responses in most CA1 neurons to exogenous NMDA applied by pressure or superfusion, with reversal on washout of acamprosate. These data suggest that acamprosate may act postsynaptically to increase the NMDA component of excitatory transmission to hippocampal CA1 pyramidal neurons. Considering the known interaction of ethanol with NMDA receptors, this acamprosate modulation of NMDA receptor-mediated neurotransmission could provide a mechanism of action underlying the clinical efficacy of acamprosate.

Acamprosate↗
PubMed1995

Reduced long-term potentiation in the dentate gyrus of transgenic mice with cerebral overexpression of interleukin-6.

The cytokine interleukin-6 (IL-6) may be a contributing mediator of CNS injury in several neurological disorders. To investigate the role of IL-6 in memory-related disorders, we examined transgenic mice (GFAP-IL6) with cerebral overexpression of IL-6 using paired-pulse facilitation, paired-pulse inhibition, and long-term potentiation (LTP) in an in vitro preparation. We found that paired-pulse potentiation and inhibition in the dentate gyrus of hippocampal slices prepared from the GFAP-IL6 mice did not differ significantly from age-matched control animals, suggesting that the increase in paired-pulse inhibition seen previously in in vivo studies of this model was due to alterations of afferents from other brain regions. However, LTP in the dentate was significantly reduced in slices from GFAP-IL6 transgenic mice when compared with littermate wild-type controls, providing support for a role of IL-6 in the pathogenesis of neurodegenerative associated memory-related disorders.

Animals↗
PubMed1995

Ethanol enhances muscarinic cholinergic neurotransmission in rat hippocampus in vitro.

Previous studies from our laboratory showed that ethanol enhances muscarinic excitatory responses in rat hippocampal neurons in vivo and, like muscarinic agonists, reduces the M-current (IM) in these neurons in vitro. Therefore, we used extracellular and intracellular recording techniques in the hippocampal slice preparation to examine the mechanisms underlying this ethanol-muscarinic interaction. Surprisingly, superfusion or local application of low concentrations of acetylcholine (ACh), carbachol (CCh) or muscarine reduced the amplitudes of CA1 field potentials evoked by stratum radiatum (SR) stimulation. This effect was blocked by 1 microM atropine but was independent of the method of agonist application, the site of application or the SR stimulus paradigm. In intracellular and extracellular single unit recordings, cholinergic depressions of field potentials were correlated with: (1) depolarization of pyramidal neurons; (2) spike discharge increases; (3) reduction of amplitudes of postsynaptic potentials and (4) reduction of late afterhyperpolarizations (AHPs). Superfusion of low ethanol concentrations (11-22 mM) alone had little effect on SR-evoked field potentials but enhanced (by 10-90%) both the depressions of evoked field potentials and depolarizations elicited by the muscarinic agonists. Ethanol (22-44 mM) also enhanced both the amplitude and duration of the muscarinic slow excitatory postsynaptic potentials (sEPSPs) recorded intracellularly in CA1 and CA3 neurons. This effect was enhanced by eserine and blocked by atropine, verifying involvement of muscarinic receptors. These results suggest that: (1) caution be used in interpreting results of field potential studies regarding drug-induced excitability changes; and (2) ethanol in just-intoxicating concentrations enhances endogenous muscarinic synaptic transmission as well as responses to exogenous muscarinic agonists.

Acetylcholine↗
PubMed1994

Ethanol inhibits glutamatergic neurotransmission in nucleus accumbens neurons by multiple mechanisms.

The nucleus accumbens (NAcc) likely plays a role in the rewarding effects of several addictive drugs such as opiates and EtOH. We showed previously that low EtOH concentrations reduced glutamatergic excitatory postsynaptic potentials (EP-SPs) in NAcc neurons. Naloxone inhibited this effect. In the present study we have begun characterizing the receptors involved in the evoked EPSPs and examined the action of EtOH on these receptors by using intracellular recording (voltage- and current-clamp) in the rat NAcc slice. At depolarized membrane potentials, we found 6-cyano-7-nitroquinoxaline-2,3-di-one-resistant EPSPs that were blocked by the N-methyl-D-aspartate (NMDA) receptor antagonist DL-2-amino-5-phosphonovalerate. In 6-cyano-7-nitroquinoxaline-2,3-dione (a non-NMDA glutamate receptor antagonist), EtOH 66 mM decreased these NMDA-EPSPs. Application of exogenous NMDA or non-NMDA [kainate, (R,S)-alpha-amino-3-hydroxy-5-methyli-soxazole-4-propionic acid or quisqualate] glutamate agonists evoked reversible depolarizations or inward currents. The NMDA-induced currents increased with membrane depolarization and were blocked by DL-2-amino-5-phosphonovalerate. EtOH 11 to 200 mM decreased the NMDA currents significantly and dose-dependently, without effect of naloxone. Higher EtOH concentrations (44-66 mM) also reduced slightly kainate-induced currents (again without a naloxone effect), but not (R,S)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid or quisqualate currents. These data suggest that NAcc core neurons express both NMDA and non-NMDA glutamate receptors. Because low EtOH concentrations reduce the EPSPs at normal resting potentials, but not responses to non-NMDA glutamate agonists, EtOH probably acts both pre- and postsynaptically: by an opioid-dependent reduction of glutamate release and by postsynaptically reducing NMDA and kainate currents. By virtue of the likely role NAcc plays in alcoholism, these actions could represent major determinants in the intoxicating and reinforcing properties of EtOH.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Voltage-dependent effects of opioid peptides on hippocampal CA3 pyramidal neurons in vitro.

Opioid peptides, and especially the dynorphins, have been localized to several circuits in the CA3 hippocampal region, yet electrophysiological studies often find mixed effects of opiates on the excitability of CA3 neurons. Reasoning that these mixed effects might involve voltage-dependent actions, we tested the effect of several opiates on CA3 pyramidal neurons using single-electrode voltage-clamp recording in a slice preparation of rat hippocampus. In most CA3 neurons, the voltage-dependent K+ current known as the M-current (IM) was uniquely sensitive to the opioid peptides, with the direction of response dependent upon the opiate type and concentration. Thus, an opiate selective for kappa receptors, U-50,488H, significantly augmented IM. The kappa-selective agonists dynorphin A and dynorphin B, which exist in mossy fiber afferents to CA3 pyramidal neurons, also markedly augmented IM at low concentrations (20-100 nM). By contrast, dynorphin A at higher concentrations (1-1.5 microM) often reduced IM. Similarly, several opiates [e.g., D-Ala2,D-Leu5-enkephalin: (DADL), [D-Pen2,5]-enkephalin (DPDPE)] known to act on the delta receptor subtypes reduced the M-current, with partial reversal of this effect by naloxone. Neither the selective mu-receptor agonist [D-Ala2, NMe-Phe4, Gly-ol]-enkephalin (DAMGO) nor the nonopioid fragment of dynorphin, des-Tyr-dynorphin, consistently altered IM. These opiate effects on IM were accompanied by changes in conductance and holding current consistent with their respective effects on IM. Dynorphin A did not measurably affect the Q-current, a conductance known to contribute to inward rectification in hippocampal pyramidal neurons. The opiate effects on IM were not altered by pretreatment with Cs+ (which blocks IQ) or Ca2+ channel blockers. The opposing effects of the dynorphins (both A and B) and DADL on IM were antagonized by naloxone (1-3 microM), and the dynorphin-induced augmentations of IM were usually reversed by the kappa receptor antagonist norbinaltorphimine. These results suggest that the opiates can have opposing effects on the same voltage-dependent K+ channel type (the M channel) in the rat CA3 pyramidal neuron, with the direction of the response depending on which receptor subtype is activated. These data not only help explain the mixed effects of opiates seen in other studies, but also suggest a potential postsynaptic function for the endogenous opiates contained in the CA3 mossy fibers.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1993

Interleukin 1 beta inhibits synaptic strength and long-term potentiation in the rat CA1 hippocampus.

Cytokines such as interleukin-1 beta (IL-1 beta) are released in the nervous system following inflammation or infection. Recently, IL-1 beta was shown to enhance synaptic inhibitory mechanisms. We therefore investigated the effect of IL-1 beta superfusion on long-term potentiation (LTP), the cellular model of memory and learning, evoked in the CA1 region by tetanic stimulation of the stratum radiatum in the rat hippocampal slice. IL-1 beta (150 pM-1.5 nM) superfused 10 min before tetanic stimulation significantly reduced LTP of the slope of the population excitatory postsynaptic potential (pEPSP) and the population spike (PS) amplitude in CA1 in a concentration-dependent manner. IL-1 beta (1.5 nM) applied for 10 min 1 h before tetanus significantly inhibited LTP of the PS amplitude and pEPSP slope and reduced pEPSP and PS values before tetanus as well, although the PS returned to control values before tetanus. Heat-inactivated IL-1 beta had no effect on pre-tetanus pEPSP or PS values or the induction of LTP. These data demonstrate that IL-1 beta modulates synaptic potentials and reduces LTP. These findings have important implications for the role of IL-1 beta in neuronal disorders following infection, perhaps best exemplified by HIV-1-associated dementia.

Animals↗
PubMed1993

Somatostatin inhibition of hippocampal CA1 pyramidal neurons: mediation by arachidonic acid and its metabolites.

We used electrophysiological methods in a slice preparation to study the mechanisms of somatostatin (SS) effects on hippocampal pyramidal neurons. SS hyperpolarizes hippocampal pyramidal neurons in part by augmenting the time- and voltage-dependent M-current (IM), which has been shown to be reduced by muscarinic agonists. The SS effects are abolished by the phospholipase A2 inhibitors 4-bromophenacyl bromide and quinacrine. Arachidonic acid (AA) mimics all the effects of SS on hippocampal pyramidal neurons. The effects of AA and SS on IM are blocked by the lipoxygenase inhibitor nordihydroguaiaretic acid but not by the cyclooxygenase inhibitor indomethacin. Prostaglandins E2, F2 alpha, and I2 do not increase IM. However, the specific 5-lipoxygenase inhibitors 5,6-methanoleukotriene A4 methylester and 5,6-dehydroarachidonic acid both blocked the IM-augmenting action of either SS or AA. Leukotriene C4 (but not leukotriene B4) increases IM to the same extent as AA. IM was not altered by the 12-lipoxygenase product 12-hydroperoxyeicosatetraenoic acid, and SS effects were not altered by the 12-lipoxygenase inhibitor baicalein. These data implicate 5-lipoxygenase metabolite(s) (probably leukotriene C4) as a mediator for the IM-augmenting effect of SS. In addition, when the IM effect is blocked by lipoxygenase inhibitors, both SS and AA elicit another outward current that is not blocked by either lipoxygenase or cyclooxygenase inhibitors, suggesting a direct role of AA itself distinct from the IM effect. SS did not alter significantly Ca(2+)-dependent action potentials or, in whole-cell recordings, inward currents likely to represent high-threshold Ca2+ currents. The combined results of these studies suggest that SS hyperpolarizes hippocampal neurons by two mechanisms, both mediated through the AA system. However, one mechanism (IM) involves a metabolite of AA and is most effective at slightly depolarized potentials, whereas the other may involve AA itself and be more effective at membrane potentials near rest.

Animals↗
PubMed1993

[The effects of opioid peptides on postsynaptic potentials in slice preparation of nucleus accumbens of rats].

Behavioral studies indicate that nucleus accumbens (NAcc) is a key brain area for the rewarding effects of opiate and for opiate addiction. To find out the cellular correlates of opiate addiction, the effects of three kinds of opiate peptides on membrane potential and postsynaptic potentials (PSPs) in a slice preparation of NAcc were studies by intracellular recording and current-clamp technique. The results indicated that superfusion of opioid 7 peptides did not affect membrane potential and input resistance of NAcc neurons, but significantly reduced postsynaptic potentials. Naloxone superfusion significantly reversed the depressant effects of mu and sigma receptor agonist (DAGO, D-Pen) on PSPs, but not on those of kappa agonist (U-50). These results suggest that opioid peptides predominantly reduce synaptic transmission in NAcc.

Animals↗
PubMed1993

Ethanol decreases glutamatergic synaptic transmission in rat nucleus accumbens in vitro: naloxone reversal.

The nucleus accumbens septi (NAcc) is a key brain region in the rewarding effects of addictive drugs such as opiates and ethanol. We recently showed that opiate peptides reduced both excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs) in NAcc neurons of a slice preparation, with naloxone (Nal) reversal (Yuan et al., 1992). To test other addictive drugs, we used intracellular recording in this rat NAcc slice preparation to investigate ethanol actions on NAcc neuronal properties. Ethanol 22 to 66 mM had little reproducible effect on membrane potential or input slope resistance, but reduced the amplitude of EPSPs evoked by stimulation of the peri-tubercle region ventral to NAcc. Ethanol 22, 44 and 66 mM all significantly decreased the EPSPs evoked by half-maximal stimulation to 80, 60 and 68% of control, respectively. Superfusion of 11 mM ethanol had no effect. To confirm a direct ethanol action on EPSPs, we tested 44 mM ethanol in the presence of 30 microM bicuculline to block IPSPs. In these cells ethanol still decreased EPSP size, suggesting GABAAergic IPSPs are not involved in this effect. The glutamate receptor blocker 6-cyano-7-nitroquinoxaline-2,3-dione abolished the EPSPs evoked at resting membrane potentials. As ethanol actions mimic those of opiates in reducing EPSPs without effect on resting membrane potentials in the NAcc, we applied the opiate antagonist Nal together with ethanol. Nal 1 to 2 microM significantly reversed ethanol (44 mM) reduction of EPSP amplitude. Thus, our data suggest that a major effect of intoxicating concentrations of ethanol in NAcc is to reduce glutamatergic synaptic transmission.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcohol Deterrents↗
PubMed1992

Concentrations of carbachol stimulating phosphoinositide hydrolysis cause a sustained decrease in membrane potential and firing rate: role of inositol and inositol polyphosphate second messengers.

We have investigated the relationship between muscarinic agonist-stimulated phosphoinositide (PI) hydrolysis and electrophysiological responses in rat hippocampal slice preparations. In a previous extracellular study, we found that muscarinic agonists at concentrations that stimulate PI hydrolysis result in a biphasic firing response; an initial increase in firing followed by loss of firing at higher concentrations. To test the hypothesis that variability in obtaining consistent loss of firing is related to depletion of intracellular inositol, we investigated the effects of adding exogenous inositol to the buffer. We now report that concentrations of inositol similar to those in cerebral spinal fluid (30-100 microM) augment carbamylcholine (carbachol, CCh) mediated loss of firing and [3H]inositol-1,3,4,5-tetrakisphosphate ([3H]Ins(1,3,4,5)P4) formation. Inhibition of firing produced by 30 microM CCh in the presence of inositol was associated with a sustained depolarization of 20-25 mV, an increased slope resistance in the depolarized range (-60 to -40 mV), and a parallel shift in the hyperpolarized (-100 to -70 mV) range of the voltage-current curve and increased frequency of spontaneous IPSPs. Under voltage-clamp, measurements of the M-current (IM) showed sustained inactivation by CCh with reversal after washout of CCh. Manual depolarization of cells by current injection to the same level of depolarization as attained with CCh did not usually lead to the same loss of firing. These findings suggest that IM, and possibly other voltage-independent currents or ion pumps, may cause loss of firing only in part through a depolarization blockade of firing and not through desensitization. Furthermore, CCh treatment without inositol did not depolarize neurons as much as CCh with inositol, and usually did not cause a delayed loss of firing. Brain slice preparations may thus require physiological concentrations of inositol to show consistent or maximum phosphoinositide-mediated electrophysiological responses.

Action Potentials↗