Alkylation of rat dopamine transporters and blockade of dopamine uptake by EEDQ.

Effects of the alkylating agent EEDQ (N-ethoxycarbonyl-2-ethoxy-1, 2-dihydroquinoline) on levels of dopamine transporter (DA(T)) and function were examined in caudate-putamen (CPu) tissue from rat brain. EEDQ produced profound, dose-dependent decreases in DA(T) binding in homogenates (IC(50)=78 microM) and frozen sections (IC(75)=200 microM) that were not reversed by washing. EEDQ also blocked uptake of [(3)H]DA in CPu synaptosomes (IC(50)=17 microM).

Olanzapine, quetiapine, and risperidone: long-term effects on monoamine transporters in rat forebrain.

Long-term effects of novel atypical antipsychotic drugs on monoamine transporters are unknown. We compared labeling of dopamine (DAT) and serotonin (SERT) transporter proteins in subregions of rat corpus striatum by quantitative autoradiography with [(3)H]2-beta-carbomethoxy-3-beta-[4'-iodophenyl]tropane ([(3)H]beta-CIT) and [(3)H]paroxetine after 28 days of continuous subcutaneous infusion of olanzapine, quetiapine, risperidone, or vehicle controls. Drug treatment did not significantly alter the abundance of either transporter type in caudate-putamen or nucleus accumbens, indicating that transporter proteins required to inactivate synaptically released dopamine and serotonin resist adaptations to long-term treatment with novel antipsychotics that affect neurotransmission by these amines.

Dopamine D(4) receptors in rat forebrain: unchanged with amphetamine-induced behavioral sensitization.

Dopamine D(2) receptors are implicated in stimulant-induced behavioral sensitization.(7,10) Studies using selective receptor antagonists also implicate the D(4) receptor, a member of the dopamine D(2)-like receptor family.(3) Accordingly, dopamine D(4) and D(2)-like receptor levels in rat forebrain were examined by computed autoradiography after repeated (+)-amphetamine treatment that induced behavioral sensitization.

Regional localization of dopamine and ionotropic glutamate receptor subtypes in striatolimbic brain regions.

Localization of dopamine (D(1)-, D(2)-like, and D(4)) and ionotropic glutamate (NMDA, AMPA, and KA) receptor subtypes within the striatolimbic forebrain remains incomplete, but basic to understanding the functional organization of this important brain region. We found that frontal cortical ablation supported colocalization of D(4) and NMDA receptors on corticostriatal afferents to caudate-putamen and nucleus accumbens in rat forebrain. Local injection of kainic acid into caudate-putamen, nucleus accumbens, or hippocampus produced massive local postsynaptic losses of D(1)- and D(2)-like, as well as NMDA, AMPA, and KA receptors, and kainic acid ablation of hippocampal-striatal projections indicated the selective expression of presynaptic NMDA and KA autoreceptors.

Comparative postnatal development of dopamine D(1), D(2) and D(4) receptors in rat forebrain.

Postnatal development of dopamine D(1), D(2) and D(4) receptors in the caudate-putamen, nucleus accumbens, frontal cortex and hippocampus was assessed in rat brain between postnatal days 7 and 60. In the caudate-putamen and nucleus accumbens, density of all three receptor subtypes increased to a peak at postnatal day 28, then declined significantly in both regions (postnatal days 35-60) to adult levels. In the frontal cortex and hippocampus, these receptors rose steadily and continuously to stable, maximal adult levels by postnatal day 60.

[3H]beta-CIT: a radioligand for dopamine transporters in rat brain tissue.

[3H]2-beta-carbomethoxy-3-beta-[4'-iodophenyl]tropane (beta-CIT) was prepared and evaluated. With rat forebrain tissue, [3H]beta-CIT showed high affinity for dopamine transporters (DAT), with selectivity for DAT over norepinephrine transporters, but not serotonin transporters, as well as DAT-stereoselectivity with beta-CIT, amphetamine and methylphenidate. Affinity and selectivity for 53 compounds assayed with [3H]beta-CIT and standard DAT radioligand [3H]GBR-12935 were highly correlated (r0.

Dopamine D4 receptors: significance for molecular psychiatry at the millennium.

Extraordinary progress has been made in the molecular, genetic, anatomical, and pharmacological characterization of dopamine D4 receptors in animal and human brain. Clarification of the neurochemical and physiological roles of these cerebral receptors is emerging. Postmortem neuropathological studies have inconsistently linked D4 receptors to psychotic disorders, and genetic studies have failed to sustain conclusive associations between D4 receptors and schizophrenia.

Effects of alkylating agents on dopamine D(3) receptors in rat brain: selective protection by dopamine.

Dopamine D(3) receptors are structurally highly homologous to other D(2)-like dopamine receptors, but differ from them pharmacologically. D(3) receptors are notably resistant to alkylation by 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ), which readily alkylates D(2) receptors. We compared EEDQ with N-(p-isothiocyanatophenethyl)spiperone (NIPS), a selective D(2)-like receptor alkylating agent, for effects on D(3) and D(2) receptors in rat brain using autoradiographic analysis.

Selective alkylatation of dopamine D2 and D4 receptors in rat brain by N-(p-isothiocyanatophenethyl)spiperone.

Effects of the D2-like receptor alkylating agent NIPS (N-[p-isothiocyanatophenethyl]spiperone) on dopamine receptors in rat brain were characterized by radioreceptor assays and quantitative autoradiography. NIPS alkylated D2 and D4 receptors concentration-dependently in brain sections and transfected cells. NIPS also alkylated both receptors dose-dependently in vivo, with no effect on dopamine D1-like or serotonin 5-HT2 receptors at a dose that occluded 75% of D2 and D4 receptors.

Neuropharmacological assessment of potential dopamine D4 receptor-selective radioligands.

Radiolabeled dopamine D4 receptor-selective agents ([3H]1-benzyl-4-[ N-(3-isopropoxy-2-pyridinyl)-N-methyl]-aminopiperidine maleate; [3 H]PNU-101958. and [125I]1-[4-iodobenzyl]-4-[ N-(3-isopropoxy-2-pyridinyl)-N-methyl]-aminopiperidine; [125I]RBI-257) were prepared and characterized. With D4.

Postnatal development of dopamine D1-like receptors in rat cortical and striatolimbic brain regions: An autoradiographic study.

Postnatal development of dopamine D1-like (D1/D5) receptors in rat caudate-putamen (CPu), nucleus accumbens (NAc), hippocampus, frontal and entorhinal cerebral cortex was assessed between postnatal days (PD) 7-60 by in vitro receptor autoradiography. Density of [3H]SCH-23390 binding to D1-like receptors increased from PD-7 to a peak at PD-28 in CPu (11-fold) and NAc (23-fold), then declined by 20-40% in both regions over PD-35-60, to adult levels. In hippocampus, frontal and entorhinal cortex, D1-like receptors increased by lesser amounts (3- to 4-fold) from PD-7 to stable, maximal adult levels at PD-60.

Postnatal development of dopamine and serotonin transporters in rat caudate-putamen and nucleus accumbens septi.

Density of dopamine transporter (DAT) and serotonin transporter (5-HTT) membrane proteins in the caudate-putamen (CPu) and nucleus accumbens (NAc) of rat brain was assessed at seven ages at postnatal days (PD) 7-60, by in vitro quantitative autoradiography. Binding of [3H]GBR-12935 (to DAT) and [3H]paroxetine (to 5-HTT) increased steadily and very similarly, from low levels at PD-7 to maximal levels, to 6-7-fold higher density at PD-60 in both regions. These findings indicate that DAT and 5-HTT follow a synchronized course of development in rat CPu and NAc.

Postnatal development of dopamine D4-like receptors in rat forebrain regions: comparison with D2-like receptors.

Development of dopamine D4-like receptors in rat caudate-putamen (CPu), nucleus accumbens (NAc), frontal cortex, hippocampus, and entorhinal cortex was assessed at seven points between postnatal days 7 and 60 by computed in vitro receptor autoradiography, and compared with dopamine (DA) D2-like receptors. Density of radioligand binding to both receptor types increased from day 7 to a peak at day 28 in caudate-putamen (D4, 3.3-fold; D2, 4.

Localization of ionotropic glutamate receptors in caudate-putamen and nucleus accumbens septi of rat brain: comparison of NMDA, AMPA, and kainate receptors.

Changes in binding of selective radioligands at NMDA ([3H]MK-801), AMPA ([3H]CNQX), and kainate ([3H]kainic acid) glutamate (GLU) ionotropic receptors in rat caudate-putamen (CPu) and nucleus accumbens (NAc) were examined by quantitative autoradiography following: 1) unilateral surgical ablation of frontal cerebral cortex to remove descending corticostriatal GLU projections, 2) unilateral injection of kainic acid (KA) into CPu or NAc to degenerate local intrinsic neurons, or 3) unilateral injections of 6-hydroxydopamine (6-OH-DA) into substantia nigra to degenerate ascending nigrostriatal dopamine (DA) projections. Cortical ablation significantly decreased NMDA receptor binding in ipsilateral medial CPu (20%), and NAc (16%), similar to previously reported losses of DA D4 receptors. KA lesions produced large losses of NMDA receptor labeling in CPu and NAc (both by 52%), AMPA (41% and 45%, respectively), and kainate receptors (40% and 45%, respectively) that were similar to the loss of D2 receptors in CPu and NAc after KA injections.

Effects of hippocampal kainic acid lesions on striatolimbic dopamine D1-, D2-, and D4-like receptors.

Changes in D1-, D2- and D4-like dopamine receptor binding in rat caudate-putamen, nucleus accumbens and hippocampus were examined by quantitative in vitro receptor autoradiography after unilateral injections of kainic acid into hippocampus to degenerate local intrinsic neurons and remove hippocampal-striatal projections. Radioligands were: [3H]SCH-23390 for D1-like (D1/D5) receptors, [3H]nemonapride alone for D2-like (D2/D3/D4) receptors and with 300 nM S[-]-raclopride and other masking agents for D4-like receptors. Kainic acid injections produced massive local hippocampal losses of D1-like (75%), D2-like (77%) and D4-like (79%) receptor binding but did not significantly alter D1-, D2- or D4-like receptor density in caudate-putamen or nucleus accumbens.

Effects of hippocampal lesions on striatolimbic ionotropic glutamatergic receptors.

Changes in ionotropic N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate glutamatergic (Glu) receptor binding in rat caudate putamen (CPu) and nucleus accumbens (NAc) and hippocampal CA1 and CA3 regions were examined after unilateral injections of kainic acid (KA) into hippocampus to degenerate local intrinsic neurons and remove the glutamatergic afferents projecting to CPu and NAc. KA injections produced large losses of NMDA, AMPA and kainate receptors in hippocampal CA1 and CA3 regions. In addition, NMDA and kainate, but not AMPA, receptors were significantly reduced in NAc and CPu.

Isomeric selectivity at dopamine D3 receptors.

Racemic 7-hydroxy-N,N-dipropylaminotetralin (7-OH-DPAT) shows greater affinity for limbic-selective dopamine D3 receptors than for more ubiquitous dopamine D2 receptors. R(+)-7-OH-DPAT was prepared and evaluated in radioreceptor assays using membranes of fibroblasts expressing the human dopamine D3 receptor as well as rat striatal membranes containing dopamine D2 receptors. This enantiomer had 2-fold greater D3 affinity than the racemate and similarly greater D3 vs.

Interactions of fluoxetine with metabolism of dopamine and serotonin in rat brain regions.

Given evidence of inhibitory effects of serotonin on dopaminergic neurotransmission, a series of experiments sought neurochemical evidence of interactions between the selective serotonin transport inhibitor fluoxetine and the metabolism of dopamine (DA) or serotonin (5-HT) in regions of rat brain that might account for extrapyramidal side-effects associated with clinical use of fluoxetine. There were significant inhibitory effects of acute or repeated fluoxetine treatment on the turnover of 5-HT (accumulation of 5-hydroxytryptophan, or ratio of [5-hydroxyindoleacetic acid]/[5-HT]) in striatum, nucleus accumbens and frontal cerebral cortex, but only minor effects on metabolism of DA (accumulation of dihydroxyphenylalanine, or [homovanillic acid]/[DA] ratio), even at high doses or with repeated treatment, and no significant inhibition of the DA metabolism-increasing actions of haloperidol.