Characterization of binding of [3H]GBR 12935 (1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl)-piperazine) to membranes and to solubilized membrane extracts from terminal field regions of mesolimbic, mesocortical and nigrostriatal dopamine pathways.

The binding characteristics of [3H]GBR 12935 (1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine), a selective dopmaine uptake inhibitor, were examined in intact membrane preparations and solubilized extracts of terminal field regions of dopamine pathways in the brain of the rats. There were many similarities in the properties of binding sites for [3H]GBR 12935 in the striatum, nucleus accumbens and olfactory tubercle. The binding of [3H]GBR 12935 was saturable and the affinity constants were not significantly different between regions of the brain. The binding of [3H]GBR 12935 was inhibited by amfonelic acid, GBR 12909, mazindol, methylphenidate and cocaine, with comparable affinities in each region of the brain and with the same order of potency in both preparations. Furthermore, the rank order of potencies for inhibiting the binding of [3H]GBR 12935 was the same as for inhibiting the uptake of [3H]dopamine in these regions of the brain. There did appear to be some degree of heterogeneity of binding sites for [3H]GBR 12935 in each of these regions of the brain, as both amfonelic acid and mazindol were best fitted by two-site models. Whether this apparent heterogeneity was due to the existence of two distinct binding sites or to two components of a single site is unclear. It did not, however, appear to be due to binding to uptake sites for norepinephrine or serotonin, as neither nisoxetine nor fluoxetine, selective inhibitors of the uptake of norepinephrine and serotonin, respectively, inhibited the binding of [3H]GBR 12935, at concentrations which inhibit the uptake of norepinephrine or serotonin.

Opioid receptor regulation of the release of norepinephrine in brain.

The ability of opioids to inhibit the release of norepinephrine (NE) from slice preparations of brain has been tested. Slices of brain were preincubated with [3H]NE allowing uptake of the [3H]NE into intraneuronal stores of NE. After rinsing, the tissues were incubated at 37 degrees C in Krebs buffer containing 5mM K+, for estimation of baseline release and then in 20 mM K+ to stimulate release. The [3H]NE released into the incubation medium was increased by blockade of neuronal re-uptake with desipramine and by blockade of alpha 2-adrenoceptors with yohimbine. These agents were used routinely in subsequent incubations. Release was also Ca2+ dependent. Stimulated release of [3H]NE from slices of cortex of the guinea pig and rat was inhibited by the mu opioid receptor agonist, Tyr-D-Ala2-Gly-NMePhe-Gly-ol (DAGO) in a naloxone-reversible manner, although naloxone itself produced a measurable inhibitory effect in the absence of opioid agonist. Stimulated release of [3H]NE from slices of guinea pig cortex was also inhibited by the delta receptor selective peptide, [D-Pen2, D-Pen5] enkephalin (DPDPE), and the kappa receptor selective agent, U50,488H. The inhibitory effect of both agents was reversed by naloxone. In rat cortex, DAGO induced a similar inhibition of release to that seen in guinea pig cortex, but DPDPE and U50,488H were much less effective, producing only weak inhibition even in large doses. Similar results were obtained when effects of opioids on [3H]NE release from hippocampus and cerebellum of the guinea pig and rat were compared. In guinea pig tissues, agonists acting preferentially through mu, delta and kappa receptors were all active in inhibiting stimulated release of [3H]NE, but in hippocampus and cerebellum of the rat, only DAGO inhibited release while DPDPE and U50,488H either had no effect or potentiated the stimulated release. These results suggest that in the rat only mu type opioid receptors mediate an inhibitory regulation of NE release from the cortex, hippocampus and cerebellum terminal projections of locus coeruleus noradrenergic neurons. In the guinea pig, stimulated release of [3H]NE was subject to inhibitory regulation by mu, delta and kappa opioid receptors.

Opioid ligand binding sites in the spinal cord of the guinea-pig.

The properties of opioid binding sites in membranes from the spinal cord of the guinea-pig were analyzed in experiments employing radiolabeled opioid ligands, selective or partially selective for mu, delta and kappa-type binding sites. Incubation was conducted at 37 degrees C in a quasi-physiological modified Krebs medium, containing sodium and magnesium. The types of binding sites were discriminated on the basis of their affinities for [3H-D-Ala2-MePhe4-Gly5-ol]enkephalin ([3H]DAGO), [3H-D-Ala2-D-Leu5]enkephalin, and [3H]ethylketocyclazocine and the relative potencies of the displacing ligands, DAGO, [D-Ser2-Leu5]enkephalyl-Thr and trans-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)- cyclohexyl]benzeneacetamide methanesulfonate hydrate (U50488H), which are selective for mu, delta and kappa type binding sites respectively. In membranes from whole spinal cord, kappa type sites comprised about 60%, mu about 30% and delta about 10% of the total of mu, delta and kappa binding sites. Binding sites of the mu type were also found in the lumbo-sacral region of guinea-pig spinal cord, in contrast to earlier reports of their absence from this tissue. Morphine showed a better than 500-fold selectivity for mu over kappa sites in spinal cord, while nalbuphine and (-)1-cyclopentyl-5-(1,2,3,4,5,6-hexahydro-8-hydroxy-3,6,11- trimethyl-2,6-methano-3-benzazocin-11-yl)3-pentanone methanesulfonate (WIN 44441-3) showed about a 10-fold selectivity for mu sites. The drug U50488H had about a 150-fold greater affinity for kappa than mu-type binding sites.

Selective opioid antagonist effects on opioid-induced inhibition of release of norepinephrine in guinea pig cortex.

Opioid agonists with selectivity for mu, delta and kappa-receptors have each been shown to inhibit the K+-stimulated release of [3H]norepinephrine (NE) from slices of guinea pig cortex maintained in vitro. In order to provide further evidence that each of these types of opioid receptor can regulate the release of NE in this tissue, experiments with receptor-type selective opioid antagonists have been conducted. In initial experiments, the selectivity of the antagonists for specific types of opioid receptors in the cortex of the guinea pig in an incubation medium of the same composition as that used for release studies was confirmed. The delta-receptor selective antagonist, ICI 174,864, prevented the inhibitory actions of the delta-selective agonist, [D-Pen2,D-Pen5]enkephalin (DPDPE), but had little effect on the inhibitory actions of the mu-selective agonist, Tyr-D-Ala-Gly-MePhe-Gly-ol (DAMGO), or the kappa-selective agonist, U-50,488H. In contrast, the kappa-selective antagonist, nor-binaltorphimine (nor-BNI) prevented the inhibitory actions of U-50,488H, but had little effect on the inhibitory actions of DPDPE or DAMGO. The greater potency of the partially mu-selective antagonist, naloxone, in reversing the effects of DAMGO relative to those of DPDPE or U-50,488H was confirmed. These results support the conclusion that mu- delta- and kappa-opioid receptors each exert a negative regulatory effect on the stimulated release of NE in the cortex of the guinea pig.

Neuropeptide Y-mediated enhancement of NMDA-stimulated [3H]dopamine release from rat prefrontal cortex is reversed by sigma1 receptor antagonists.

Sigma (sigma) receptors are located in limbic areas, including the prefrontal cortex, where decreased dopamine levels have been linked to negative symptoms. Although the endogenous ligands for sigma receptors are unknown, neuropeptide Y (NPY) has been named as the potential endogenous agonist at these receptors. NPY enhanced NMDA-stimulated [3H]dopamine release in rat prefrontal cortex. This was in contrast to the inhibition produced by the sigma agonists (+)pentazocine and BD737. However, four sigma antagonists, including one which is sigma1 selective, that reverse (+)pentazocine- or BD737-mediated inhibition all reversed the NPY-mediated enhancement. In addition, PYX-1, a Y receptor antagonist, reversed both the (+)pentazocine- and BD737-mediated inhibition and the NPY-mediated enhancement of release. Peptide YY (PYY), [Leu31,Pro34]NPY and NPY(13-36) did not mimic the effect of NPY. Our findings are consistent with NPY acting as an endogenous ligand for a subtype of sigma receptor with characteristics different from Y1, Y2 and Y3 receptors but sensitive to PYX-1. These findings suggest a role for NPY, via sigma receptors, as a modulator of dopamine levels in the prefrontal cortex.

Regulation of [3H] dopamine release from mesolimbic and mesocortical areas of guinea pig brain by sigma receptors.

The role of sigma (sigma) receptors in brain function is poorly defined. They are located in limbic areas, including nucleus accumbens (NAC) and prefrontal cortex (PFC), both of which are thought to be involved in schizophrenia. Many antipsychotics (APs), including haloperidol, bind with high affinity to sigma receptors. Dopaminergic hyperactivity in NAC is thought to underlie positive symptoms of schizophrenia, while dopaminergic hypoactivity in PFC is thought to underlie negative symptoms. Sigma receptors regulate N-methyl-D-aspartate (NMDA)-stimulated [3H] dopamine ([3H]DA) release in caudate-putamen (CP), the neuroanatomical substrate for extrapyramidal side effects resulting from chronic AP treatment. In the current study, we investigated whether sigma receptors could similarly regulate DA release in mesolimbic and mesocortical tissue, and the relative participation of different sigma receptor subtypes in this process. We found that, in NAC, regulation of DA release by the prototypical sigma agonist (+)pentazocine was mediated predominantly by the sigma 1 receptor, whereas in the PFC a portion of the (+)pentazocine effect was likely mediated by the sigma 2 receptor. We also observed, in both the NAC and PFC, that regulation of DA release by the sigma agonist BD737 was mediated primarily by the sigma 1 receptor. In addition, we determined that (+)pentazocine or BD737 effects on DA release were not mediated via opioid receptors, nor the phencyclidine (PCP) binding site within the NMDA receptor-operated cation channel, nor by sigma receptor effects upon [3H]DA accumulated by noradrenergic terminals in PFC.

The sensitivity of opioid receptor types to regulation by sodium and GTP.

The effects of Na+ and GTP on agonist binding to mu, delta, and kappa opioid binding sites in membranes from guinea pig cortex have been studied. All incubations were in a modified Krebs-Hepes buffer at 37 degrees; effects of Na+ were evaluated by replacement with an equal concentration of K+. mu binding sites appeared more sensitive than delta sites, while kappa sites were the least sensitive to both regulators. However, even at kappa sites, Na+ induced a significant reduction in agonist affinity and GTP caused a significant reduction in maximum binding.

Neurotensin, N-acetyl-aspartylglutamate and beta-endorphin modulate [3H]dopamine release from guinea pig nucleus accumbens, prefrontal cortex and caudate-putamen.

Dopaminergic hyperactivity in nucleus accumbens and dopaminergic hypoactivity in prefrontal cortex are thought to underlie positive and negative symptoms of schizophrenia, respectively. The caudate putamen is the neuroanatomical substrate for extrapyramidal side effects resulting from chronic antipsychotic treatment. We sought to identify potential endogenous regulators of dopamine release that might produce differential effects in these brain areas. We tested neurotensin, N-acetyl-aspartyl-glutamate and beta-endorphin for potential regulation of [3H]dopamine release in these regions of guinea pig brain. All three peptides stimulated dopamine release, above basal activity, at all concentrations tested in the three regions. Neurotensin significantly enhanced and N-acetyl-aspartyl-glutamate had no significant effect on N-methyl-D-aspartate-stimulated release from all three regions. In contrast, beta-endorphin significantly inhibited N-methyl-D-aspartate-stimulated release in nucleus accumbens and caudate putamen. These results suggest that these neuropeptides may regulate endogenous dopamine release and therefore may be potential therapeutic targets for antipsychotic drug development.

SH-SY5Y cells as a model for sigma receptor regulation of potassium-stimulated dopamine release.

Previous studies in our laboratory using rat brain tissue have shown that neuropeptide Y (NPY) can enhance NMDA- and potassium-stimulated dopamine release from various brain regions and that this enhancement is reversed by sigma (sigma) receptor antagonists. In the current study, we sought to determine whether SH-SY5Y cells are suitable for investigating sigma receptor effects and whether any sigma receptors present are of the subtype responsive to NPY. We compare mechanisms by which the prototypical sigma receptor agonist (+)-pentazocine, and the proposed endogenous sigma receptor ligand NPY regulate potassium-stimulated [(3)H]dopamine release from SH-SY5Y cells. Both (+)-pentazocine and NPY inhibit potassium-stimulated [(3)H]dopamine release. Unlike our studies in rat brain tissue, the effect of NPY on [(3)H]dopamine release is not reversed by sigma receptor antagonists. SH-SY5Y cells appear to be an appropriate model to study the regulation of dopamine release by sigma receptors or by NPY receptors, but this population is not identical to that population identified in brain slices.

Differential modulation of NMDA-stimulated [3H]dopamine release from rat striatum by neuropeptide Y and sigma receptor ligands.

Although the identity of the endogenous ligands for sigma (sigma) receptors is unknown, neuropeptide Y (NPY) has been named as a possible candidate for a natural transmitter at these receptors. Using a superfusion system, we compared the effect of NPY on NMDA-stimulated [3H]dopamine release in rat striatum to that of the sigma agonists (+)-pentazocine and BD737. In contrast to (+)-pentazocine- or BD737-mediated inhibition of release, NPY enhanced release. However, the same sigma antagonists (BD1008, DuP734, haloperidol and DTG) that reverse (+)-pentazocine- or BD737-mediated inhibition, as well as a Y receptor antagonist, PYX-1, all reversed the enhancement. PYX-1 also reversed the (+)-pentazocine- and BD737-mediated inhibition of release. Peptide YY (PYY) and [Leu31,Pro34]NPY did not mimic the effect of NPY. NPY13-36 enhanced release to the same extent as NPY but the effect was not reversed by sigma antagonists. Our findings are consistent with the potential role of NPY as an endogenous ligand for a subtype of sigma receptor with characteristics different from Y1, Y2 and Y3 receptors but sensitive to PYX-1.

Evidence for differential localization of two binding sites for L-[3H]glutamate in rat fascia dentata.

Two binding sites for L-[3H]glutamate were tentatively localized in rat fascia dentata by determining the effects of selective lesions on specific binding. Both destruction of dentate granule cells with colchicine and ablation of the ipsilateral entorhinal cortex markedly reduced radioligand binding to a quisqualate-sensitive site (GLU A), but only the entorhinal lesion significantly reduced binding to a site that is less sensitive to quisqualate (GLU B). These results suggest that GLU A binding sites are localized mainly on the dentate granule cells, whereas GLU B binding sites are localized, in part, on the perforant path fibers, but not on granule cells.

Sigma receptor regulation of norepinephrine release from rat hippocampal slices.

Multiple sigma receptor subtypes have been identified in the hippocampus, yet their physiological role remains largely undefined. In the current study, we examined the role of sigma receptors in the regulation of N-methyl-D-aspartate (NMDA)-stimulated [3H]norepinephrine ([3H]NE) release from rat hippocampal slices. Both sigma agonists (+)pentazocine and BD737 inhibited stimulated norepinephrine release in a concentration-dependent manner. The sigma1 antagonist DuP 734 completely antagonized the inhibition of release by all concentrations of BD737 tested. However, DuP 734 only partially reversed inhibition of release by (+)pentazocine concentrations above 100 nM. 1,3 Di-o-tolylguanidine (DTG), but not haloperidol, antagonized BD737-mediated inhibition of release. DTG also completely antagonized inhibition of release by 100 nM (+)pentazocine yet haloperidol produced only a partial reversal. A combination of DuP 734 and haloperidol produced complete reversal of (+)pentazocine-mediated inhibition, suggesting potential involvement of multiple sigma receptor subtypes in the regulation of norepinephrine release. Both (+)pentazocine and BD737 failed to inhibit stimulated release in the presence of tetrodotoxin, suggesting that sigma receptors regulating NE release are not located on noradrenergic nerve terminals. These results suggest that sigma receptors may be a therapeutic target for disorders resulting from noradrenergic imbalance in hippocampus.

Comparison of the effects of cocaine and other inhibitors of dopamine uptake in rat striatum, nucleus accumbens, olfactory tubercle, and medial prefrontal cortex.

It is thought that inhibition of dopamine reuptake into neurons may play a major role in the mechanisms by which cocaine produces its reinforcing effects. The striatum, while rich in dopamine terminals, is not implicated in drug reinforcement, whereas the mesolimbic dopamine pathway appears to play a primary role. It is therefore possible that the properties and drug sensitivities of the dopamine uptake systems in the nigrostriatal, mesolimbic, and mesocortical tracts differ. The effects of cocaine, GBR 12909, amfonelic acid, and methylphenidate on dopamine uptake in the striatum, nucleus accumbens, olfactory tubercle, and medial prefrontal cortex were examined. Over 80% of the dopamine uptake in each of the 4 regions was sodium-dependent and exhibited Km values of approximately 100 nM. Cocaine, GBR 12909, amfonelic acid, and methylphenidate each biphasically inhibited uptake in the striatum, nucleus accumbens and olfactory tubercle with GBR 12909 and amfonelic acid being approximately 50-fold more potent than cocaine or methylphenidate. In the medial prefrontal cortex, cocaine and GBR 12909 could inhibit only about 40% of the [3H]dopamine uptake. There are similarities in the properties and drug sensitivities of the dopamine uptake systems in brain areas which are implicated in drug reinforcement and those which are not.

The role of L-type voltage dependent calcium channels in stimulated [3H]norepinephrine release from canine hippocampal slices following global cerebral ischemia and reperfusion.

The hippocampus is among those brain regions which are selectively vulnerable to ischemic damage. Hippocampal damage due to transient cerebral ischemia is mainly of the delayed, non-necrotic type which may arise after disruption or activation of specific cellular systems, including transmitter release through excitatory amino acid receptors. We investigated the contribution of L-type voltage dependent calcium channels (VDCCs) to glycine (GLY) potentiated N-methyl-D-aspartate (NMDA) receptor- and potassium-stimulated [3H]norepinephrine (NE) release in a canine model of global cerebral ischemia and reperfusion. Tissue was collected from four experimental groups: non-arrested controls (NA), global cerebral ischemia induced by 10 minute cardiac arrest (CA), and CA followed by 30 min or 24 hours reperfusion after restoration of spontaneous circulation. Brain slices prepared from all groups accumulated approximately equivalent amounts of [3H]NE. The sensitivity of [3H]NE release to stimulation by NMDA/GLY or elevated potassium was unchanged after ischemia and reperfusion. About 30% of release stimulated by the addition of 20 mM potassium was inhibited by the NMDA receptor-operated channel antagonist MK801 in all groups except CA in which only 4% of release was inhibited by MK801. The ability of 1 microM nitrendipine (NTP) to block stimulated release indicated that the contribution of the L-type VDCC to potassium or NMDA/GLY-stimulated release was significant only in NA and 24 hour reperfused animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Dopamine release from canine striatum following global cerebral ischemia/reperfusion.

The elevation of extracellular dopamine (DA) levels in the striatum of experimental animals subjected to ischemic insult has been well documented. The contribution of excessive DA to neuronal damage can be inferred from the ability of DA antagonists, as well as selective destruction of dopaminergic tracts, to confer neuroprotection in models of ischemia. In the current study, we report an enhanced releasability of preloaded [3H]DA in response to either elevated potassium or N-methyl-D-aspartate (NMDA) from striatal slices of beagles that had experienced 10 min of ischemia induced by cardiac arrest. The elevation in sensitivity to potassium stimulation was transient, approaching control levels after 30 min of reperfusion. In contrast, release stimulated by NMDA was elevated immediately after cardiac arrest and remained elevated for as long as 24 h of reperfusion. Release stimulated by NMDA was enhanced by glycine (Gly) and inhibited by MK801, consistent with mediation through the NMDA receptor/channel complex. The increased sensitivity of DA release, coupled with the high levels of excitatory amino acids (EAAs), including glutamate (Glu), aspartate (Asp) and Gly in ischemic brain, probably contribute to the extensive neuronal cell damage.

Selective and reversible increase in the number of quisqualate-sensitive glutamate binding sites on hippocampal synaptic membranes after angular bundle kindling.

The specific binding of L-[3H]glutamate to hippocampal synaptic membranes was examined in rats kindled by tetanic stimulation of the angular bundle. One day after the last of a minimum of 3 class 4 kindled seizures, the binding of L-[3H]glutamate to a quisqualate-sensitive site (GLU A) was about 40% greater than in electrode-implanted unstimulated controls. Saturation binding data indicated an increase in the maximum density of GLU A binding sites with no change in their affinity for L-glutamate. No such increase was detected 28 days after the last kindled seizure, although the animals were still kindled. Radioligand binding to a site that is much less sensitive to quisqualate (GLU B) was unaffected by kindling stimulation. These observations suggest that an increase in GLU A binding sites could be involved in the induction, but not in the maintenance, of the kindled state.

Evidence that the sigma(1) receptor is not directly coupled to G proteins.

Sigma (sigma) receptors have been implicated in psychosis, cognition, neuroprotection, and locomotion in the central nervous system. The signal transduction mechanisms for sigma receptors have not been fully elucidated. In this study, we examined the possible coupling between sigma(1) receptors and heterotrimeric guanine nucleotide-binding proteins (G proteins) in rodent brain. In sigma(1) receptor-rich cerebellar membrane preparations, the competitive binding curves of two sigma(1) agonists, (+)pentazocine and 1S,2R-(-)-cis-N-[2-(3, 4-dichlorophenyl)ethyl]-N-methyl-2-(1-pyrrolidinyl)cyclohexylamine (BD737), were unaffected by the addition of 10 microM guanosine-5'-O-(gamma-thio)-triphosphate (GTPgammaS). Neither (+)pentazocine (1-100 microM) nor BD737 (0.01-10 microM) stimulated GTPase activities significantly above basal levels in agonist-stimulated GTPase activity assays in cerebellar membranes. Furthermore, when using the method of agonist-stimulated [35S]GTPgammaS binding as assessed by autoradiography, we did not observe significant stimulation of [35S]GTPgammaS binding in rat brain sections by either (+)pentazocine or BD737. The above results demonstrate that the sigma(1) receptor is not likely be directly coupled to G proteins.

Phencyclidine and dizocilpine modulate dopamine release from rat nucleus accumbens via sigma receptors.

Phencyclidine (PCP) binds to many sites in brain, including PCP receptors located within the N-methyl-D-aspartate (NMDA) receptor-operated cation channel and sigma (sigma) receptors. In this study, we compare mechanisms by which PCP, dizocilpine (MK-801), the prototypical sigma receptor agonist (+)-pentazocine, and the proposed endogenous sigma receptor ligand neuropeptide Y regulate potassium (K(+))-stimulated [3H]dopamine release from slices of rat nucleus accumbens. (+)-Pentazocine inhibits K(+)-stimulated [3H]dopamine release, and neuropeptide Y enhances it. Both effects are blocked by sigma(1) and neuropeptide Y receptor antagonists, suggesting possible inverse agonism at a subpopulation of sigma/neuropeptide Y receptors. In contrast, PCP and MK-801 both enhance K(+)-stimulated [3H]dopamine release via sigma(1) and sigma(2) receptor subtypes, as demonstrated by antagonist sensitivity. Regulation of release by both (+)-pentazocine and neuropeptide Y persists in the presence of tetrodotoxin suggests that the sigma/neuropeptide Y receptors mediating the modulation are located presynaptically on dopaminergic nerve terminals, but tetrodotoxin eliminates regulation by PCP and MK-801, suggesting that receptors mediating their effects are located upstream from dopaminergic nerve terminals.

sigma1 Receptors in rat striatum regulate NMDA-stimulated [3H]dopamine release via a presynaptic mechanism.

The role of the sigma1 receptor in the regulation of N-methyl-D-aspartate (NMDA)-stimulated [3H]dopamine release from rat striatal slices was examined. The sigma receptor agonist 1S,2R-(--)-N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(1-pyrrolidinyl)cy clohexylamine (BD737) inhibited stimulated release in a concentration-dependent manner. The sigma1 receptor antagonist, 1-(cyclopropylmethyl)-4-(2'-(4"-fluorophenyl)-2'-oxoethyl)piperidi ne HBr (DuP 734), reversed inhibition of release by BD737. Haloperidol, di-o-tolylguanidine (DTG) and N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(1-pyrrolidinyl)ethylamine (BD1008) reversed the BD737-mediated inhibition of release. Haloperidol and DTG also antagonized inhibition of stimulated release by (+)-pentazocine. Furthermore, BD737 and (+)-pentazocine inhibited stimulated release in the presence of tetrodotoxin, suggesting that sigma1 receptors regulating dopamine release are located on dopaminergic nerve terminals. These data suggest that sigma1 receptors may be important in the regulation of glutamate-stimulated dopamine release.

Regulation of [3H]norepinephrine release from guinea pig hippocampus by sigma2 receptors.

The binding profile of the sigma2 receptor ligand endo-N-(8-methyl-8-azabicyclo[3.2.1.]oct-3-yl)-2,3-dihydro-(1-methyl)eth yl-2-oxo-1H-benzimidazole-1-carboxamidehydrochloride (BIMU-8) had previously been determined, but its agonist/antagonist status at sigma2 receptors had not been identified. We therefore investigated the effects of BIMU-8 for its ability to regulate the stimulated release of [3H]norepinephrine from slices of guinea pig hippocampus. BIMU-8 alone, at a concentration chosen to occupy 50% of sigma2 receptors, had no significant effect on N-methyl-D-aspartate (NMDA)-stimulated release of [3H]norepinephrine. We have shown previously that the sigma receptor agonist (+)-pentazocine inhibits NMDA-stimulated release in a concentration-dependent manner, producing a biphasic inhibition curve. Similarly, the sigma receptor agonist 1S,2R-(-)-cis-N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(1-pyrrolidinyl )cyclohexylamine (BD737) produced a broad inhibition curve. The inhibition by low concentrations of (+)-pentazocine or BD737 that selectively activated sigma1 receptors was reversed by the sigma1-selective receptor antagonist (1-(cyclopropylmethyl)-4-2'-oxoethyl)piperidine HBr (DuP 734). In the current study, when the sigma1 component of inhibition by (+)-pentazocine was blocked by DuP 734, the remaining component of inhibition mediated by sigma2 receptors was reversed by BIMU-8. Our results suggest that (1) BIMU-8 is an antagonist at sigma2 receptors and that (2) sigma2 receptors contribute to regulation of norepinephrine release in guinea pig hippocampus.