Chronic treatment with antipsychotic drugs does not alter G protein alpha or beta subunit levels in rat brain.

Groups of rats received once daily subcutaneous treatments for 22 days with haloperidol (0.5 mg/kg), clozapine (20 mg/kg), SCH 23390 (0.2 mg/kg) or vehicle. Quantitative immunoblots for Gi alpha 1, Gi alpha 2, G(o alpha), Gs alpha (45 kD), G beta 35 and G beta 36 were performed on membranes from the following brain regions: striatum, nucleus accumbens, substantia nigra, ventral tegmental area (VTA), prefrontal cortex and hippocampus (CA1). No significant alterations were found in the levels of any of these G protein subunits, in any brain region in treated vs control rats.

Differential receptor reserve for 5-HT1A receptor-mediated regulation of plasma neuroendocrine hormones.

Treatment of rats with the serotonin 5-HT1A agonist 8-hydroxy-2-(di-n- propylamino)tetralin (8-OH-DPAT, 1 mg/kg s.c.) markedly elevated plasma levels of corticosterone (CORT), adrenocorticotropic hormone (ACTH) and prolactin (PRL); the levels of growth hormone were unaffected. Pretreatment with the irreversible receptor antagonist N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ, 6 mg/kg s.c.) greatly attenuated the increase in plasma CORT produced by 8-OH-DPAT (0.3 mg/kg s.c.). Prevention of EEDQ-induced 5-HT1A receptor inactivation by prior treatment with the reversible mixed 5-HT1A/beta-adrenergic antagonist (+/-)pinodolol (30 mg/kg s.c.) blocked the reduction of the CORT response to 8-OH-DPAT. In contrast, prevention of EEDQ-induced inactivation of 5-HT2, alpha-1- and alpha-2-adrenergic and D1 and D2 dopamine receptors by a cocktail of selective antagonists of these receptors did not block the attenuation of the CORT response to 8-OH-DPAT. Dose-response curves were obtained for 8-OH-DPAT (0.01-3 mg/kg s.c.)-induced elevation of plasma CORT, ACTH and PRL after treatment (24 hr earlier) with vehicle or EEDQ (6 mg/kg s.c.) and analyzed for the extent of receptor reserve. Whereas substantial receptor reserves were observed for the 8-OH-DPAT rise in plasma CORT (80%) and ACTH (50%), no receptor reserve was seen for the increase in plasma PRL. The results are discussed with regard to potential differences in the receptors, G proteins, effectors and/or stoichiometric relationships between these components of the signal transduction pathway, leading to elevation of these plasma hormones after treatment with 8-OH-DPAT.

Evidence that striatal synthesis-inhibiting autoreceptors are dopamine D3 receptors.

The activation constants (KA; dose required to occupy 50% of receptors) for reversal of gamma-butyrolactone (GBL)-induced elevation of striatal L-3,4-dihydroxyphenylalanine (L-DOPA) levels via stimulation of presynaptic dopamine receptors were determined for apomorphine and two dopamine D3 receptor-selective agonists, quinpirole and LY163502 (quinelorane). The KA values correlated significantly with the affinities (Ki) of the agonists for the D3 (r = 0.999, P < 0.05) but not the D2 (r = -0.13) receptor, suggesting that striatal synthesis-inhibiting autoreceptors are of the D3 rather than the D2 subtype.

Extracellular dopamine concentration in the retina of the clawed frog, Xenopus laevis.

Dopamine reaches targets in the outer retina of the clawed frog (Xenopus laevis) by diffusion from a network of dopaminergic cells and processes located predominantly at the junction of inner nuclear and inner plexiform layers. We obtained values for the steady-state release, uptake, and extracellular concentration of dopamine in the retina by a combination of HPLC (with electrochemical detection), scintillation spectroscopy, and fast-scan cyclic voltammetry. Vitreal concentrations of dopamine varied from 564 +/- 109 nM in light-adapted eyes near the time of subjective dawn to 156 +/- 12 nM in dark-adapted eyes. The data are consistent with a simple model for steady-state dopamine diffusion from an appropriately sited thin-sheet source. This model was used to generate a profile of extracellular dopamine concentration as a function of retinal depth. The model predicted an increase in the dopamine concentration from the vitreous to the layer of dopaminergic cells, remaining constant from that layer to the distal tips of the photoreceptors. This prediction was borne out by comparing fast-scan voltammetric measures of dopamine at the distal tips of the receptors with the vitreal concentrations determined by HPLC using electrochemical detection.

A novel method for rapid enrichment of lactotrophs from dispersed anterior pituitary cells of the rat.

A polyclonal antibody to the rat D2 dopamine (DA) receptor was rapidly and covalently attached to surface-activated polystyrene cultureware (MicroCEL-Lector plates). Addition of a suspension of dispersed rat anterior pituitary cells resulted in the rapid (within 1 h) selection of cells possessing D2 DA receptors (i.e. lactotrophs). Four-fold enrichment (from about 20% in the suspension to about 80%) was routinely obtained, as judged by prolactin (PRL) immunostaining. The enriched cells were virtually free of fibroblasts and were much more homogeneous in appearance than untreated cells after 3 days in culture. Lactotroph-enriched cell cultures displayed similar functional characteristics as untreated cells when assessed by determining dose-response curves for inhibition of PRL secretion by the DA agonist N-propylnorapomorphine. This method may be generally applicable for the selective enrichment and purification of desired cell types from heterogeneous mixtures in tissue dispersions.

Comparative effects of chronic 8-OH-DPAT, gepirone and ipsapirone treatment on the sensitivity of somatodendritic 5-HT1A autoreceptors.

Chronic treatment with the full 5-hydroxytryptamine1A (5-HT1A) agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), either by twice daily subcutaneous injection (0.2 or 2.0 mg/kg) or continuous subcutaneous administration via osmotic minipumps (0.4 or 4.0 mg/kg/day), for 14 days, had no effect on the dose-response curves for inhibition of 5-hydroxytryptophan (5-HTP) accumulation in rat cortex or hippocampus by 8-OH-DPAT or the partial 5-HT1A agonist BMY 7378. In contrast, chronic treatment with the nonbenzodiazepine putative anxiolytic gepirone via osmotic minipumps (20 mg/kg/day) resulted in a small but significant rightward shift (1.8-fold) in the dose-response curve to 8-OH-DPAT challenge in the cortex and a slightly larger shift (2.4-fold) in the hippocampus. Similarly, chronic treatment with another putative anxiolytic, ipsapirone, administered via twice daily subcutaneous injections (20 mg/kg), also resulted in a rightward shift (2.6-fold) in the dose-response curve to 8-OH-DPAT in the cortex and a slightly smaller shift (2.3-fold) in the hippocampus. Neither drug, however, decreased the maximal response. The present results are consistent with the suggestion that the clinical anxiolytic effects of gepirone and ipsapirone, and not of 8-OH-DPAT, may be related to their ability to desensitize somatodendritic 5-HT1A autoreceptors; other potential mechanisms are discussed.

Comparative effects of receptor inactivation, 17 beta-estradiol and pertussis toxin on dopaminergic inhibition of prolactin secretion in vitro.

Akin to receptor inactivation with phenoxybenzamine (PBZ) (1 microM, 1 hr), treatment of anterior pituitary cells with 17 beta-estradiol (10 nM, 3 days) right-shifted the dose-response curve for inhibition of prolactin (PRL) secretion by the full agonist R-(-)-N-n-propylnorapomorphine (NPA) and reduced the maximal effect [EC50 (pM) and percent maximal effect: control, 25.4 and 81.2; PBZ, 115.3 and 57.9; 17 beta-estradiol, 358 and 58.6]. PBZ treatment of 17 beta-estradiol-pretreated cultures further reduced the maximal response but did not alter the EC50. Plots of receptor occupancy vs. response indicated a large receptor reserve for NPA (approximately 60%) in control cultures but its abolition by 17 beta-estradiol. 17 beta-Estradiol pretreatment elicited identical rightward shifts (4.5-fold) and similar reductions in maximal PRL inhibition by quinpirole and (+)-3-PPP, although these drugs were partial agonists with dissimilar efficacies relative to NPA (0.61 and 0.12, respectively) at presynaptic striatal D2 receptors. However, receptor inactivation experiments with (+)-3-PPP and quinpirole, and subsequent comparison of receptor occupancy vs. response plots, demonstrated that the relative efficacies of quinpirole and (+)-3-PPP were reversed in the striatum and anterior pituitary. In striatum, half-maximal response to quinpirole and (+)-3-PPP required 6.2 and 30% receptor occupancy, respectively, whereas 25.6 and 9.6% occupancy was required in the pituitary. Pertussis toxin treatment (10 ng/ml, 24 hr) produced large shifts in the dose-response curves for all three agonists (8.4-21.9-fold), but was distinguished from the effects of both PBZ and 17 beta-estradiol by a significant (P < .001) decrease in the slope factor.(ABSTRACT TRUNCATED AT 250 WORDS)

Serotonin 5-HT1A receptor-mediated hypothermia in mice: absence of spare receptors and rapid induction of tolerance.

The mixed 5-hydroxytryptamine1A (5-HT1A) receptor agonist/antagonist 8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspirol-[4.5]- decane-7,9-dione (BMY 7378) (5 mg/kg) did not significantly depress body temperature, but pretreatment with BMY 7378 blocked hypothermia induced by the selective 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). In contrast, another partial 5-HT1A agonist, pindolol (10 mg/kg), slightly but significantly depressed body temperature by itself but did not attenuate hypothermia elicited by 8-OH-DPAT. Attempts to identify the synaptic locus of the receptor were unsuccessful because depletion of central serotonin (5-HT) by treatment with para-chlorophenylalanine (PCPA; 3 x 150 mg/kg) did not alter the hypothermic response to 8-OH-DPAT. Partial, irreversible 5-HT1A receptor inactivation by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) (1 mg/kg) reduced the maximal hypothermic effect of 8-OH-DPAT (to 53% of control) without altering its ED50 (0.96 mg/kg). Analysis of the data indicated a linear relationship between 5-HT1A receptor occupancy and hypothermic response, that is, absence of receptor reserve. When groups of mice were treated with each of five different doses of 8-OH-DPAT (0.04, 0.16, 0.63, 2.5, and 10 mg/kg) 48 h apart, there was a significant reduction in hypothermic response after the second injection, but only at the three highest doses. The results demonstrate that 8-OH-DPAT-induced hypothermia in mice is mediated by a 5-HT1A receptor whose synaptic localization is uncertain but that has no receptor reserve. In addition, tolerance is observed after only a single agonist treatment.

The effects of pertussis toxin on dopamine D2 and serotonin 5-HT1A autoreceptor-mediated inhibition of neurotransmitter synthesis: relationship to receptor reserve.

Irreversible inactivation of striatal D2 dopamine (DA) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) or inactivation of striatal guanine nucleotide binding proteins (G proteins) with pertussis toxin (PT) shifted the dose-response curve for N-n-propylnorapomorphine (NPA)-mediated inhibition of gamma-butyrolactone (GBL)-induced elevation of L-3,4-dihydroxyphenylalanine (L-DOPA) to the right, with a decrease in the maximum response. For the partial agonist (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine [(+)-3-PPP], in contrast, there was little shift in the ED50, after inactivation of either D2 receptors or G proteins. Completely analogous effects were found at the somatodendritic 5-HT1A autoreceptor in the raphe nuclei, mediating inhibition of the synthesis of serotonin (5-HT); the full agonist, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and the partial agonist, buspirone were utilized to inhibit the synthesis of 5-HT, as measured by changes in levels of L-5-hydroxytryptophan (5-HTP). Additionally, in both systems, combined treatment with pertussis toxin, followed by EEDQ, reduced the maximum effect, when compared to either agent alone but had little further effect on the ED50. In systems exhibiting a large receptor reserve for agonists, such as those described above, the same pattern of response seen after inactivation of receptors or G proteins may reflect the operation of a common mechanism underlying the phenomenon of receptor reserve.

Receptor reserve for 5-hydroxytryptamine1A-mediated inhibition of serotonin synthesis: possible relationship to anxiolytic properties of 5-hydroxytryptamine1A agonists.

The irreversible receptor antagonist N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) was used to determine the relationship between receptor occupancy and response at central 5-hydroxytryptamine1A (5-HT1A) serotonin receptors mediating the inhibition of serotonin synthesis in rat cortex and hippocampus. Rats were treated with vehicle or EEDQ (2 or 6 mg/kg) and 24 hr later dose-response curves were constructed for inhibition of 5-hydroxytrytophan (5-HTP) accumulation (after decarboxylase inhibition with NSD-1015) by the selective 5-HT1A agonists 8-hydroxy-2(di-n-propylamino)tetralin (8-OH-DPAT) (0.01-3 mg/kg), buspirone (0.1-7.5 mg/kg), and ipsapirone (0.1-6.25 mg/kg) and the 5-HT1A agonist/antagonist BMY 7378 (0.015-5 mg/kg). In vehicle-pretreated rats, a similar maximal inhibition of 5-HT synthesis (range, 52-59%) was observed in both brain areas with 8-OH-DPAT, buspirone, and ipsapirone. These three agonists were also more potent in reducing 5-HTP accumulation in the cortex than in the hippocampus (ED50, 8-OH-DPAT, 14 and 30 microgram/kg; buspirone, 0.42 and 0.63 mg/kg; ipsapirone, 0.44 and 1.26 mg/kg, respectively). In the cortex, EEDQ treatment shifted the dose-response curves for 8-OH-DPAT, buspirone, and ipsapirone 8.6-, 2.0-, and 2.8-fold to the right, respectively. Corresponding rightward shifts in the hippocampus were smaller, 6.0-, 1.6-, and 2.1-fold, respectively. The EEDQ-induced shifts in the dose-response curves were accompanied by reductions in maximal response. In contrast, whereas the maximal inhibition of cortical 5-HTP accumulation by BMY 7378 (55%) was similar to that obtained with the agonists, maximal response in the hippocampus was much smaller (32%). Furthermore, in both brain regions EEDQ reduced the maximal response to BMY 7378 without shifting the dose-response curves. Analysis of the data by the double-reciprocal method of Furchgott, followed by calculation of fractional receptor occupancy for each dose of agonist, revealed a nonlinear relationship between receptor occupancy and response for 8-OH-DPAT, buspirone, and ipsapirone in both brain regions, demonstrating the presence of a large receptor reserve. For BMY 7378, in contrast, linear relationships were obtained. Because 5-HT1A receptor-mediated regulation of 5-HT synthesis appears to be mediated by somatodendritic autoreceptors on 5-HT neurons in the midbrain raphé nuclei, the results suggest that these autoreceptors possess a large receptor reserve for agonists. The relevance of these findings for the mechanism of action of nonbenzodiazepine anxiolytics is discussed.

Involvement of D1 and D2 Dopamine Receptors in the Control of Horizontal Cell Electrical Coupling in the Turtle Retina.

We studied the actions of D1 and D2 dopamine agonists and antagonists on the coupling of horizontal cell axons in the turtle retina by a combination of pharmacological and electrophysiological methods. Both D1 and D2 receptors were identified in membrane fractions by radioligand binding using [3H]-SCH 23390 and [3H]-spiperone, respectively. The KD of both receptor classes were identical (0.21 nM) but D1 receptor density exceeded that of D2 receptors by more than four-fold. D1 agonists increased the activity of adenylate cyclase in a dose-dependent manner, whereas D2 agonists were without significant effect by themselves, nor did D2 antagonists block the D1-mediated increase in adenylate cyclase activity. Intracellular recordings and Lucifer Yellow dye injections were used to characterize the modifications of the receptive field profile of horizontal cell axons (H1AT) exposed to different pharmacological agents. Dopamine or D1 agonists (0.05 - 10 microM) induced a marked constriction of the H1AT receptive field, whereas D2 agonists elicited a small expansion of the receptive field. However, in the presence of a D1 antagonist, as well as IBMX to inhibit phosphodiesterase, D2 agonists (10 - 70 microM) induced a marked increase in the receptive field profile. These results indicate that both D1 and D2 dopamine receptors play a role in shaping the receptive field profile of the horizontal cell axon terminal in the turtle retina.

Behavioral recovery after irreversible inactivation of D-1 and D-2 dopamine receptors.

Irreversible inactivation of both D-1 and D-2 dopamine (DA) receptors by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) resulted in complete loss of stereotypy response to R-(-)-N-propylnorapomorphine (NPA; 0.1-1.0 mg/kg, s.c.) 24 hr later. Stereotyped sniffing recovered much more rapidly than oral behaviors. The D-2 antagonist sulpiride (200 mg/kg) and the putatively nonselective antagonist cis-flupenthixol (2 mg/kg), administered prior to EEDQ, prevented the loss of NPA-induced sniffing but only partially protected against loss of oral behaviors 24 hr later. Complete protection of both behaviors was seen after pretreatment with a combination of sulpiride and the selective D-1 antagonist SCH 23390 (1 mg/kg); pretreatment with the selective D-1 antagonist SCH 23390 alone, however, did not modify the rate of recovery of either behavioral response. The results suggest that either different populations of DA receptors mediate expression of these behaviors or stimulation of a small fraction of the total DA receptor pool may be sufficient to elicit sniffing but not oral responses. Furthermore, maintaining a normal complement of D-2 rather than D-1 receptors appears to be a critical determinant for the elicitation of these behaviors.

Absence of spare autoreceptors regulating dopamine agonist inhibition of tyrosine hydroxylation in slices of rat striatum.

Incubation of rat striatal slices with forskolin (0.05-10 microM) elicited a dose-dependent increase in the activity of tyrosine hydroxylase (TH) assayed in subsequently solubilized extracts of the enzyme. At low concentrations (33 microM) of the cofactor (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride TH activity was increased 2.5 to 3-fold. Kinetic analysis of TH activity as a function of (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride concentration indicated that the enzyme isolated from control slices was composed of multiple species with different Km's for cofactor. Treatment with forskolin (1.5-15 microM) converted the enzyme into a single species with a low Km (28 microM) for (6R)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride. The dopamine (DA) agonist R-(-)-N-n-propylnorapomorphine (0.1 microM) reversed forskolin-induced activation of TH. Concentration-response curves were obtained for inhibition of forskolin-stimulated TH by R-(-)-N-n-propylnorapomorphine and the DA autoreceptor-selective agonists (+)- and (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine and 3-[4-(4-phenyl-1,2,3,6-tetrahydropyridyl-1)-butyl]indole. R-(-)-N-n-propylnorapomorphine maximally inhibited forskolin-stimulated activity 85%, as indicated by ALLFIT computer analysis of concentration-response curves. (+)-3-(3-hydroxyphenyl)-N-n-propylpiperidine and 3-[4-(4-phenyl-1,2,3,6-tetrahydropyridyl-1)-butyl]indole produced a lower degree of maximal inhibition (54 and 63%, respectively), whereas (-)-3-(3-hydroxyphenyl)-N-n-propylpiperidine was inactive. The D2 DA receptor blocker sulpiride (1 microM) competitively antagonised the effects of all the agonists.(ABSTRACT TRUNCATED AT 250 WORDS)

Enhancement by the D1 dopamine agonist SKF 38393 of specific components of stereotypy elicited by the D2 agonists LY 171555 and RU 24213.

Dose-effect curves were obtained for five specific components of stereotyped behavior (locomotor hyperactivity, rearing, sniffing, grooming and oral activity) elicited by the nonselective dopamine agonists N-propylnorapomorphine (NPA; 0.001-4.5 mg/kg) and apomorphine (APO; 0.3-10.0 mg/kg), the selective D2 agonists LY 171555 (0.1-18.0 mg/kg) and RU 24213 (0.7-20.0 mg/kg) and the selective D1 agonist SKF 38393 (0.6-40.0 mg/kg) in male Sprague-Dawley rats. All the agonists except SKF 38393 elicited dose-dependent sniffing. Both NPA and APO produced robust oral activity at high doses, with concomitant reductions in other behaviors. Neither RU 24213 nor SKF 38393 elicited dose-dependent oral behavior, whereas LY 171555 induced a maximal level of oral activity which was much less intense than that produced by NPA or APO. SKF 38393 alone induced only dose-dependent grooming. Co-treatment of Sprague-Dawley rats with SKF 38393 (5-40 mg/kg) and either LY 171555 (0.1-1.6 mg/kg) or RU 24213 (6.7 mg/kg) produced significant enhancement of sniffing but not of oral activity. Significant enhancement of both behavioral components was observed in male Wistar rats. Differences in specific components of behavioral response on treatment with combinations of D1 and D2 agonists may reflect use of animal strains with different ratios of D1/D2 innervation and/or different behavioral criteria.

Relationship between receptor occupancy and response at striatal dopamine autoreceptors.

The irreversible dopamine (DA) receptor antagonist N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) was used to determine the extent of receptor reserve at DA autoreceptors regulating in vivo tyrosine hydroxylase activity. Rats were treated with vehicle or EEDQ (1 X 0.5-2 X 6 mg/kg, subcutaneously) and, 24 hr later, dose response curves were generated for DA agonist reversal of gamma-butyrolactone-induced striatal L-3,4-dihydroxyphenylalanine (L-DOPA) accumulation. Double reciprocal plots were obtained of equieffective doses of agonist required to elicit response at several levels of effect before and after partial irreversible receptor inactivation. A pseudo-dissociation constant (pseudo-KA, in units of dose) and the fraction of receptors remaining active (q) were determined; these values were then used to calculate the relationship between receptor occupancy and response. The ED50 (1 microgram/kg) for the full DA receptor agonist N-propylnorapomorphine (NPA) was shifted 2.8, 4.8-, and 11.3-fold to the right after partial irreversible receptor blockade which left the fraction of receptors remaining active (q) at 0.37, 0.17 and 0.058, respectively. Corresponding maximal reversal of L-DOPA accumulation was 100, 77, and 58%, indicating a nonlinear relationship between receptor occupancy and response for NPA and the presence of a large receptor reserve; maximal and half-maximal responses were calculated to require occupancy of 30 and 3.8% of the total receptor pool, respectively. Dose response curves were also obtained for the DA autoreceptor-selective agents EMD 23,448 and (+)- and (-)-3-PPP before and after EEDQ treatment. In controls, EMD 23,448 and (+)-3-PPP, like NPA, completely reversed striatal gamma-butyrolactone-induced L-DOPA accumulation, whereas the maximal effect of (-)-3-PPP was 52% reversal. After EEDQ treatment (6 mg/kg), EMD 23,448 and (+)-3-PPP showed relatively small shifts in ED50 values. Furchgott analysis demonstrated that all three atypical agents are partial agonists at the DA autoreceptor with efficacies of 0.19 (EMD 23,448), 0.12 [(+)-3-PPP], and 0.05 [(-)-3-PPP] relative to NPA. The presence of a larger receptor reserve at pre-versus postsynaptic D2 DA receptors and the partial agonist character of drugs such as EMD 23,448 and the enantiomers of 3-PPP may account for their autoreceptor selectivity.

Receptor reserve at striatal dopamine autoreceptors: implications for selectivity of dopamine agonists.

The dose response curve for apomorphine reversal of gamma-butyrolactone (GBL)-induced L-DOPA accumulation in rat striatum was shifted almost 6-fold to the right after partial irreversible blockade (83%) of dopamine (DA) autoreceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ); however, the maximal response was not reduced. In contrast, the major effect of a similar degree of irreversible blockade (86%) on the dose-response curve for the autoreceptor-selective agent EMD 23,448 was a reduction in maximal response (60% of control), indicating that EMD 23,448 is a partial agonist. A large receptor reserve therefore exists at the DA autoreceptor, which may explain in part why many DA agonists are more potent in models pre- than postsynaptic receptor activation.

Inactivation of D1 and D2 dopamine receptors by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline in vivo: selective protection by neuroleptics.

Treatment of rats with the peptide coupling agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (6 mg/kg i.p.) irreversibly reduced the binding of [3H]spiperone ([3H]SPIP) and cis-[3H] piflutixol to striatal D2 and D1 receptors, respectively, by 70 to 75%. In each instance only the receptor density was affected, without a change in the dissociation constant (Kd) of either radioligand. Pretreatment with sulpiride (200 mg/kg i.p.), a selective D2 antagonist, preferentially protected [3H]SPIP sites against N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline-induced inactivation, whereas pretreatment with SCH 23390 (3 mg/kg i.p.), a putative selective D1 antagonist, preferentially blocked the inactivation of cis-[3H]piflutixol binding sites. N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline markedly reduced radioligand binding to cortical alpha-1 ([3H]prazosin) and alpha-2 [( 3H]yohimbine) receptors (10-20% of control) but had a lesser effect on serotonin-2 ([3H]SPIP) and serotonin-1 ([3H]5-HT) receptors (30-40% of control). Muscarinic cholinergic ([3H] quinuclidinyl benzilate) and beta adrenergic ([3H]dihydroalprenolol) receptors were only slightly affected. None of these nondopaminergic sites were protected by sulpiride or SCH 23390, with the exception of serotonin-2 and serotonin-1 which were partially protected by the latter. SPIP (0.2 mg/kg i.p.), haloperidol (1 mg/kg i.p.) and pimozide (2 mg/kg i.p.) all selectively protected the D2 receptor, whereas cis-flupenthixol (2 mg/kg i.p.) protected both dopamine receptors; its inactive isomer trans-flupenthixol (20 mg/kg i.p.) protected neither. Bulbocapnine (25 mg/kg s.c.) selectively, but partially, protected the D1 site.(ABSTRACT TRUNCATED AT 250 WORDS)

Chronic haloperidol does not alter agonist affinity for dopamine receptors in vitro.

Agonist competition for [3H]spiperone binding to striatal dopamine D2 receptors was studied in rats rendered supersensitive by chronic treatment with haloperidol. The classical dopamine agonist (-)-N-n-propylnorapomorphine displaced [3H]spiperone biphasically, with IC50 values of 0.5 and 140 nM for the high and low affinity components, respectively. Neither the relative density nor the affinity of either site for (-)-N-propylnorapomorphine was affected by chronic haloperidol treatment. On the other hand, the novel agonist EMD 23 448 displaced [3H]spiperone monophasically. Although this agent only displays potent dopaminergic agonism in supersensitive animals, chronic treatment with haloperidol likewise did not alter the affinity of this drug for [3H]spiperone binding sites. The results suggest that the enhanced in vivo potency of certain agonists in supersensitive animals is probably not mediated by changes in D2 receptor affinity.

Differential effects of chronic clorgyline and amfonelic acid on desensitization of striatal dopamine receptors.

Chronic treatment of rats with the MAOI clorgyline significantly reduced the density (Bmax) of cortical beta-adrenergic receptors but did not alter either the Bmax or dissociation constant (Kd) of 3H-spiperone binding to striatal DA receptors. Clorgyline co-treatment also did not significantly affect either behavioral supersensitivity to apomorphine or the increase in 3H-spiperone binding induced by chronic haloperidol. In contrast, repeated treatment with the DA uptake inhibitor amfonelic acid elicited behavioral subsensitivity and reduced striatal 3H-spiperone binding. Furthermore, amfonelic acid co-treatment prevented haloperidol-induced behavioral and receptor binding changes. The possible relevance of these findings in relation to drug choice in clinical trials of receptor sensitivity modification are discussed.

Chronic haloperidol does not increase specific dopamine receptor binding in rat frontal cortex.

The specific D2 dopamine receptor antagonist sulpiride was used to quantitate the minor component of [3H]spiroperidol binding (at 0.1 nM) to dopamine receptors in rat frontal cortex. Chronic treatment of rats with haloperidol, 0.5 mg/kg for 3 weeks or 2.5 mg/kg for 5 weeks, did not alter the specific binding of [3H]spiroperidol to dopamine receptors in frontal cortex, whereas both treatments significantly increased binding to striatal dopamine receptors. These findings may be relevant to previous studies demonstrating that the frontal cortex, but not the striatum, is resistant to the development of tolerance to the dopamine metabolite-elevating effects of chronic neuroleptic treatment.