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I Creese

Publications and source records attributed to I Creese.

At least 37 records · Page 2Linked to original sources
PubMed1991

N-(p-isothiocyanatophenethyl)spiperone, a selective and irreversible antagonist of D2 dopamine receptors in brain.

N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), an irreversible and nonselective protein-modifying reagent, has been used extensively in studies involving inactivation of receptors. Here, we present N-(p-isothiocyanatophenethyl)spiperone (NIPS), a novel and highly selective irreversible inactivator of D2 but not D1 receptors. In in vitro studies, NIPS exhibited an apparent Ki of 10 nM for [3H]methylspiperone binding to D2 receptors in rat striatum. Preincubation of the striatal membranes with NIPS followed by extensive washing resulted in up to an 80% reduction of the D2 receptor maximum binding (Bmax). Coincubation with the D2 receptor antagonist domperidone could protect against this reduction. NIPS was additionally shown to irreversibly inactivate D2 receptor binding activity in cultured cells expressing the D2 receptor protein. In in vivo administration studies, using [3H]SCH 23390 and [3H]spiperone to assay D1 and D2 receptors in vitro, 24 hr after injection (s.c.) with 5 to 40 mg/kg of NIPS D2 receptor, Bmax was decreased by 58 to 76%, without a change in D2 receptor affinity. In contrast, there was no effect on D1 receptor Bmax or affinity. There was also a small (24%) reduction in frontal cortex 5-hydroxytryptamine2 receptors by 20 mg/kg of NIPS. However, there was no effect on alpha-1 or alpha-2 adrenergic receptors in the frontal cortex, or on muscarinic cholinergic or 5-hydroxytryptamine1A receptors in the hippocampus. After single doses of either 20 mg/kg of NIPS or 10 mg/kg of EEDQ, the D2 receptor recovery rate was much slower after NIPS (half-time of receptor recovery = 170 hr) than after EEDQ (half-time of receptor recovery = 76.7 hr).(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic alpha-Antagonists↗
PubMed1990

Lack of effect of chronic dopamine receptor blockade on D2 dopamine receptor mRNA level.

The chronic treatment of rats with antipsychotic drugs which increase striatal D2 dopamine receptors did not measurably increase the level of D2 receptor mRNA indicating that the observed increase in receptors was not primarily due to transcriptional regulation. These and other data suggest that the molecular mechanisms involved in receptor regulation in response to denervation or neurotransmitter depletion may be different from those which occur in response to chronic receptor blockade by antagonist drugs.

Adrenergic alpha-Antagonists↗
PubMed1989

Comparison of two putatively selective radioligands for labeling central nervous system beta-adrenergic receptors: inadequacy of [3H]dihydroalprenolol.

[3H]Dihydroalprenolol ([3H]DHA) has been used extensively in receptor binding studies to measure beta-adrenergic receptors in the central nervous system. Usually, nonspecific binding has been defined by high concentrations of the beta-adrenergic receptor agonist isoproterenol or antagonists such as alprenolol or propranolol. Scatchard plots of such "specific" [3H]DHA saturation data in rat cerebral cortex membranes are linear. However, computer analysis demonstrated that the competition curves of these drugs for 2.0 nM [3H]DHA binding are biphasic, with a continuous inhibition of [3H]DHA binding in the concentration range usually used to determine nonspecific binding. These data indicate that another saturable high affinity site was being labeled by the radioligand and that the definition of nonspecific binding with any of these unlabeled drugs is not satisfactory. We used the nonlinear, least squares, curve-fitting program LIGAND to analyze total [3H]DHA binding, allowing the program to mathematically define nonspecific binding as a function of 3H-ligand concentration. Significantly lower Bmax (-44%) and Kd (-58%) values for beta-adrenergic receptors were found, indicating that under normal experimental procedures (defining [3H]DHA non-specific binding with these nonradioactive drugs) a second binding site was being labeled. We found that [3H]DHA binding to this site could be inhibited by drugs such as RU24969, a 5-hydroxytryptamine1A (5HT1A) and 5HT1B receptor subtype-selective agonist, and CGS12066B, a 5HT1B receptor subtype-selective agonist, which were able to compete for 15-20% of [3H]DHA binding in the nanomolar concentration range, whereas drugs that are selective for other serotonin receptor subtypes inhibited [3H]DHA binding only at much higher concentrations. Another beta-adrenergic receptor antagonist radioligand, [3H]CGP-12177, was found to be more selective for beta-adrenergic receptors. Alprenolol competition curves for [3H]CGP-12177 binding were monophasic and saturation curves, with nonspecific binding defined either by 10 microM alprenolol or by LIGAND, yielded Bmax values close to those obtained with [3H]DHA when its nonspecific binding was defined by LIGAND. [3H]DHA cannot be considered a suitable radioligand to quantify central nervous system beta-adrenergic receptors in the manner in which it has been typically used.

Alprenolol↗
PubMed1989

Reevaluation of the regulation of beta-adrenergic receptor binding by desipramine treatment.

Treatment of rats with desipramine (DMI) has been shown to down-regulate beta-adrenergic receptor-stimulated adenylate cyclase and reduce the Bmax of beta-adrenergic receptors in some brain areas. Recent reports have indicated that the down-regulation in the number of beta-adrenergic receptors following DMI treatment does not occur if the serotonin system has been impaired following parachlorophenylalanine (PCPA) or 5,7-dihydroxytryptamine injection. We have previously shown that [3H]dihydroalprenolol ([3H]DHA), the most commonly used radioligand to measure central nervous system beta-adrenergic receptors, labels another site under normal experimental procedures, in addition to the beta-adrenergic receptors. This second site has some pharmacological characteristics of the 5-hydroxytryptamine1A receptor. The depletion of serotonin following PCPA injection was indeed able to prevent the down-regulation of [3H]DHA binding sites after DMI injection. However, PCPA alone increased the density of [3H]DHA binding sites. If the nonlinear, least squares, curve-fitting program LIGAND was allowed to define [3H]DHA nonspecific binding or if the more selective beta-adrenergic receptor radioligand [3H]CGP-1277 was used, the Bmax of beta-adrenergic receptors was not changed after PCPA injection. Importantly, PCPA did not prevent beta-adrenergic receptor down-regulation following DMI treatment. The blockade of 5-hydroxytryptamine2 receptors, via ketanserin administration, during DMI treatment did not change the response of beta-adrenergic receptors. Furthermore, if LIGAND was used to define the nonspecific binding of [3H]DHA, the down-regulation of beta-adrenergic receptors was significant 24 hr after a single DMI injection. The same rapid down-regulation was demonstrated with [3H]CGP-12177. However, if [3H]DHA was used to label beta-adrenergic receptors in the "typical" manner (nonspecific binding defined by 10 microM alprenolol), a decrease in the number of beta-adrenergic receptors was significant only after seven daily DMI injections. These data demonstrate that the use of [3H]DHA to measure beta-adrenergic receptors can be misleading, because changes in its second binding site can conceal the changes occurring in beta-adrenergic receptors. Moreover, these results suggest that a similarity in the time course of action of DMI cannot be used to support the hypothesis that its therapeutic antidepressant action is related to beta-adrenergic receptor down-regulation.

Animals↗
PubMed1989

Irreversible and quaternary muscarinic antagonists discriminate multiple muscarinic receptor binding sites in rat brain.

The maximal number of binding sites (Bmax) of [3H]quinuclidinyl benzilate (QNB) binding was greater than the Bmax of N-[3H]methylscopolamine (NMS) binding to homogenates of rat brain. The competition of NMS for [3H]QNB demonstrated that NMS discriminates multiple muscarinic binding sites. Similarly, pirenzepine competition of [3H]QNB binding also revealed multiple muscarinic binding sites. Pirenzepine competition for [3H]NMS also was shallow and demonstrated the presence of binding sites with similar affinities to those labeled by [3H]QNB. These data were consistent with the presence of at least three populations of muscarinic binding sites with similar affinities for [3H]QNB: the M1 and M2 binding sites having high and low affinity for pirenzepine, respectively, but which cannot be discriminated by [3H]NMS, and a third site with high affinity for [3H]QNB which has low affinity for NMS. The classical muscarinic antagonists, atropine and scopolamine, also appear to have slightly different affinities for the putative M1 and M2 binding sites. The use of the irreversible antagonists, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) and propylbenzilylcholine mustard (PBCM), were used to elucidate the distinct properties of these multiple muscarinic binding sites. Both PBCM and EEDQ irreversibly decreased the Bmax of [3H]QNB and [3H]NMS binding in cortex. PBCM did not appear to discriminate putative M1 and M2 binding sites but selectively alkylated the high affinity NMS and QNB binding sites. In contrast, EEDQ modified the low affinity NMS binding sites such that they still bound [3H]QNB but their affinity for other muscarinic antagonists was reduced.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Age-related differential recovery rates of rat striatal D-1 dopamine receptors following irreversible inactivation.

We report here the effects of age on the steady state levels of [3H]flupentixol-labeled striatal D-1 dopamine receptors and their recovery rates following irreversible blockade in mature (4 months old) and senescent (28 months old) male Fischer 344 rats. Senescent rats exhibited significantly reduced (-22%) Bmax levels of D-1 dopamine receptors compared with their mature counterparts. In addition, the time course of recovery of D-1 dopamine receptors following irreversible receptor inactivation by a single injection of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) was significantly slower in senescent animals. This slower recovery rate was the result of decreases in both receptor production rate and receptor degradation rate constants compared with mature rats. Although EEDQ irreversibly inactivated D-1 dopamine receptors, it was without effect on dopamine uptake sites. A chronic, 21 day reserpine treatment did not significantly alter the Bmax nor the rate of recovery of D-1 dopamine receptors with respect to values in age-matched non-reserpine-treated rats. These data indicate that both the steady state levels of striatal D-1 dopamine receptors and the 'turnover' of these receptors are significantly affected as a consequence of natural aging.

Adrenergic alpha-Antagonists↗
PubMed1988

Chronic treatment with dopamine receptor antagonists: behavioral and pharmacologic effects on D1 and D2 dopamine receptors.

Rats were treated for 21 d with the selective D1 dopamine receptor antagonist SCH23390, the selective D2 dopamine receptor antagonist spiperone, the nonselective dopamine receptor antagonist cis-flupentixol, or a combination of SCH23390 and spiperone. In addition, a group of rats received L-prolyl-L-leucyl-glycinamide (PLG) for 5 d after the 21 d chronic spiperone treatment. Chronic treatment with SCH23390 resulted in a significant increase in D1 dopamine receptor density with no change in the D2 dopamine receptor density. Conversely, spiperone treatment resulted in a significant increase in D2 dopamine receptors and no change in D1 dopamine receptor density. PLG treatment had no effect. SCH23390 plus spiperone treatment resulted in a significant increase in both D1 and D2 dopamine receptor densities. However, although in vitro cis-flupentixol has an equal affinity for D1 and D2 dopamine receptors, only the D2 dopamine receptor density increased after chronic treatment with cis-flupentixol. In vivo treatment with the protein-modifying reagent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), which irreversibly inactivates D1 and D2 dopamine receptors, was used to investigate the paradoxical, selective D2 dopamine receptor up-regulation induced by cis-flupentixol treatment. In vivo treatment with cis-flupentixol before EEDQ administration prevented the D1 and D2 dopamine receptor reductions induced by EEDQ. However, cis-flupentixol protected, in a dose-dependent manner, a greater percentage of D2 dopamine receptors than of D1 dopamine receptors from EEDQ-induced modification. These data indicate that, in vivo, cis-flupentixol preferentially interacts with D2 dopamine receptors and could explain why only D2 dopamine receptors were up-regulated following chronic treatment with cis-flupentixol. Rats were tested for their cataleptic response to the administered drug over the course of the chronic drug treatment. Catalepsy scores of rats receiving spiperone decreased over the course of treatment, with a significant reduction in catalepsy occurring by treatment day 5. The profound catalepsy observed in rats receiving SCH23390 did not change over the 21 d of treatment. Rats receiving cis-flupentixol demonstrated tolerance to its cataleptogenic effects, with a significant reduction in catalepsy observed by treatment day 7. During the 3 week treatment, the time between drug injection and a full cataleptic response to cis-flupentixol increased from 20 to 60 min, suggesting a tolerance to the D2, but not D1, dopamine receptor antagonism by cis-flupentixol.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1987

The D1 dopamine receptor antagonist SCH 23390 increases cocaine self-administration in the rat.

Rats trained to intravenously self-administer cocaine hydrochloride on a fixed ratio-5 schedule were subjected to a series of systemic injections of the D1 dopamine receptor selective antagonist SCH 23390 and the D2 dopamine receptor selective antagonist spiperone. SCH 23390 produced a dose-dependent increase in cocaine intake at doses of 5, 10 and 20 micrograms/kg, but the D2 antagonist failed to reliably increase responding except at a dose of 10 micrograms/kg. These results suggest that the D1 antagonist may be more effective at blocking mesolimbic dopamine activity, and that selective D1 receptor activation may be an important component of psychostimulant reward.

Animals↗
PubMed1987

Dopamine receptor subtype imbalance in schizophrenia.

We have investigated the radioligand binding properties of D1 and D2 dopamine receptors in postmortem brains from schizophrenic patients. Consistent with previous reports, the schizophrenic population demonstrated a significant 56% increase in D2 dopamine receptor density. Importantly, the D1 dopamine receptor density was significantly reduced by 43%. These alterations in dopamine receptor densities resulted in a highly significant difference in the ratio of D2/D1 dopamine receptors between schizophrenic patients and controls. A correlation between D1 dopamine receptor density and age was apparent in the schizophrenic patients: D1 dopamine receptor density decreased markedly with age and the linear regressions of D1 dopamine receptor density versus age in both the controls and schizophrenic patients had similar slopes. These results may have clinical implications for the treatment of schizophrenia and tardive dyskinesia.

Adult↗
PubMed1987

Differential serotonin2 receptor recovery in mature and senescent rat brain after irreversible receptor modification: effect of chronic reserpine treatment.

We report here the differential time course of recovery of [3H]ketanserin-labeled serotonin2 (5-HT2) receptors in frontal cortex of mature (4 months old) and senescent (28 months old) male Fischer 344 rats. We provide evidence that the irreversible modification of 5-HT2 serotonin receptors by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline occurs at the ligand binding site and that this treatment does not appear to affect the interaction of these receptors with their guanine nucleotide regulatory protein. Senescent rats exhibited significantly reduced (-20%) maximum receptor density levels of 5-HT2 receptors compared with their mature counterparts. The time course of recovery of [3H] ketanserin binding to 5-HT2 receptors after irreversible receptor modification by a single peripheral injection of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline was significantly slower in senescent rats. The slower recovery was a function of decreases in both the receptor production rate and the degradation rate constant in senescent rats compared with mature rats. Interestingly, chronic reserpine treatment resulted in a significant decrease in control maximum receptor density values in both mature (-17%) and senescent (-18%) rats. Although the receptor production rate and receptor degradation rate constants in both mature and senescent reserpinized rats were slightly decreased, there was no significant change in the overall rates of receptor recovery when compared with their respective age-matched nonreserpinized counterparts. These data indicate that both the steady state levels and "turnover" of 5-HT2 receptors are decreased in senescence and that these receptors can be down-regulated by chronic reserpine treatment.

Adrenergic alpha-Antagonists↗
PubMed1987

Differential recovery rates of rat D2 dopamine receptors as a function of aging and chronic reserpine treatment following irreversible modification: a key to receptor regulatory mechanisms.

The density of D2 dopamine receptors identified by 3H-spiperone binding was significantly lower in the striatum of senescent (28-month-old) than in mature (4-month-old) Fischer 344 rats. The time course of recovery of 3H-spiperone binding to D2 dopamine receptors following irreversible receptor modification by a single injection of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) was significantly slower in senescent than in mature rats. The kinetics of D2 dopamine receptor repopulation could be adequately described by a model assuming a constant rate of receptor production and a rate of degradation that was dependent on receptor concentration. Both the production rate and degradation rate constant of D2 dopamine receptors were lower in the senescent rats compared to mature rats. The changes in receptor density that occur as a function of development and aging are the result of changes in both the receptor production rate and receptor degradation rate constant, which, while both are reduced, tend to maintain receptor density. That is, receptor density decreased by 26% from 4 to 28 months of age, although the receptor production rate and degradation rate constant decreased by 40-50%. Chronic treatment of rats with reserpine produced a 21% increase in the density of striatal D2 dopamine receptors of mature, but not senescent, rats, accompanied by an increase in the initial recovery rate of 3H-spiperone binding sites following EEDQ treatment. It might have been expected that the reduced formation of agonist-bound receptors would have lead to a decrease in the receptor degradation rate constant.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1987

Differential modification of striatal D1 dopamine receptors and effector moieties by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline in vivo and in vitro.

Both in vivo and in vitro treatments with the irreversible protein-modifying reagent, N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), were used to investigate rat striatal D1 dopamine receptor/effector interactions. Peripherally administered EEDQ markedly reduced D1 dopamine receptor binding and D1 dopamine receptor-stimulated adenylate cyclase in a dose-dependent manner. However, EEDQ administered in vivo did not result in functional modification of either the guanine nucleotide-regulatory protein (Ns) or the catalytic subunit of striatal adenylate cyclase as assessed via guanine nucleotide- or forskolin-stimulated cAMP production. Interestingly, the loss in D1 dopamine receptor binding did not correlate directly with observed reductions in dopamine-stimulated adenylate cyclase activity; 40% of D1 dopamine receptor binding was lost with no significant reduction in the Vmax of dopamine-stimulated adenylate cyclase activity. Conversely, the reduction by EEDQ of the adenylate cyclase activity stimulated by the partial agonist SKF38393 was reduced in parallel with EEDQ-induced reductions in the D1 dopamine receptor Bmax. However, when SKF38393-stimulated adenylate cyclase activity was potentiated by forskolin, approximately 30% of receptors could be lost with no significant reduction in cAMP production, resembling the pattern observed utilizing the full agonist dopamine. In vivo pretreatment with the specific D1 antagonist, SCH23390, prevented reductions in dopamine-stimulated adenylate cyclase activity and D1 dopamine receptor binding, suggesting that EEDQ acts at the ligand recognition site of the receptor. Unlike in vivo treatment, in vitro EEDQ treatment resulted in dose-dependent decreases in catalytic subunit activity as assessed by forskolin-stimulated cAMP production, indicating that, in vitro, the adenylate cyclase catalytic subunit is vulnerable to EEDQ-induced modification. These data indicate that EEDQ is an effective tool for elucidating the mechanisms and biochemistry of D1 dopamine receptor/effector coupling.

Adenylyl Cyclase Inhibitors↗
PubMed1986

"Selective" D-1 and D-2 receptor antagonists fail to differentially alter supersensitive locomotor behavior in the rat.

The dopamine receptor antagonists SCH 23390 and spiperone show highly selective in vitro affinity for D-1 and D-2 dopamine receptor subtypes, respectively. We studied the effects of these selective antagonists on the supersensitive locomotor response to apomorphine in rats following 6- hydroxydopamine (6OHDA) lesions of the nucleus accumbens (N. Acc.). Both D-1 and D-2 receptor antagonists produced dose-dependent blockade of the supersensitive locomotor response at doses that did not depress baseline locomotor activity. The behavioral properties of these D-1 and D-2 receptor antagonists were further examined using a simple step-down motor task. Both antagonists produced catalepsy as evidenced by dose-dependent increases in step- down latency. These results indicate that drugs with distinct in vitro dopamine binding affinities cannot be distinguished on the basis of their ability to inhibit supersensitive locomotor activity or simple motor tasks in rats in vivo.

Animals↗
PubMed1986

Effects of aging and cholinergic deafferentation on putative muscarinic cholinergic receptor subtypes in rat cerebral cortex.

Despite a 34% decrease in the activity of choline acetyltransferase (ChAT) in the rat cerebral cortex following lesions of the nucleus basalis, there were no changes in the Bmax of the antagonist ligands [3H]quinuclidinyl benzilate ((-)-[3H]QNB) or (-)-[3H]N-methylscopolamine ((-)-[3H]NMS). Furthermore, this treatment produced no significant change in the proportions or affinities of muscarinic receptors having high and low affinity for pirenzepine or (-)-NMS. These data indicate that putative M2 muscarinic receptors are not restricted to ChAT-containing neurons in rat cerebral cortex. In senescent compared to mature rats there was no significant loss of ChAT activity although a significant reduction in the Bmax of both (-)-[3H]QNB and (-)-[3H]NMS binding was observed. However, no changes in the competition of pirenzepine or (-)-NMS for the remaining (-)-[3H]QNB binding sites were observed. Therefore, there is no evidence for any differential regulation of either putative muscarinic receptor subtype in response to cholinergic deafferentation or as a function of the natural aging process.

Acetylcholine↗
PubMed1986

Functional recovery of D1 dopamine receptor-mediated stimulation of rat striatal adenylate cyclase activity following irreversible receptor modification by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ): evidence for spare receptors.

We report here the functional relationship between the time-dependent recovery of [3H]SCH 23390-labeled D1 dopamine receptors and the D1 receptor-mediated stimulation of rat striatal adenylate cyclase activity following irreversible receptor modification by in vivo administration of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. Initial decreases in receptor density (-93%) and receptor-mediated enzyme activity (-78%) were accomplished without concomitant changes in guanosine triphosphate or forskolin-stimulated enzyme activity. The percentage of maximal D1 receptor-mediated enzyme activity was significantly greater than that of D1 receptor density at all recovery times. Dopamine-stimulated enzyme activity returned to control values by day 4, although D1 receptor density remained significantly below control levels at this time. No differences in the EC50's for dopamine stimulation of enzyme activity were observed at any of the recovery times. These data demonstrate that the stoichiometric relationship between the recovering D1 dopamine receptors and D1 receptor-mediated enzyme activity is not one to one, providing evidence for the presence of 'spare' D1 dopamine receptors in rat striatum.

Adenylyl Cyclases↗
PubMed1986

Guanine nucleotide regulation of agonist interactions at [3H]SCH23390-labeled D1 dopamine receptors in rat striatum.

We report here the regulation of agonist interactions with [3H]SCH23390-labeled D1 dopamine receptors in rat striatum. Scatchard analyses of [3H]SCH23390 saturation data revealed a single high affinity binding site (KD = 0.49 nM) with a Bmax of 64 pmol/g tissue. The specific binding of 0.25 nM [3H]SCH23390 represented 90% of total binding. Antagonist competition for [3H]SCH23390 binding was monophasic (i.e. pseudo-Hill slope approximately 1) and the rank order of antagonists' affinities was consistent with the pharmacology of D1 dopamine receptors (e.g. cis-flupentixol greater than haloperidol greater than spiperone). In contrast, agonist competition curves were shallow (pseudo-Hill slope less than 1) and computer-assisted analysis indicated that, for all agonists, the data best fit a two-site model composed of a high (KH) and a low (KL) affinity component. In the presence of 0.3 mM GTP, the high affinity binding component (%RH) of various agonists was reduced by approximately 50%. No significant effect of 0.3 mM GTP on [3H]SCH23390 binding was observed. Additionally, it was noted that [3H]SCH23390 labels S2 serotonin receptors in extrastriatal brain regions. However, [3H]SCH23390 apparently does not have an affinity high enough to label S2 receptors at the concentration of [3H]SCH23390 employed in labeling striatal D1 dopamine receptors. These data indicate that [3H]SCH23390 represents a superior radioligand for labeling the two-state striatal D1 dopamine receptor in that its high percent specific binding makes it especially suitable for detailed mechanistic studies of this receptor.

Animals↗