Age-related changes of dopamine sensitive cyclic AMP generation in the rat frontal cortex

The dopamine (DA) D-1 and D-2 receptors coupled to 3',5'-cyclic adenosine monophosphate (cAMP) generation were studied in membrane particles of the frontal cortex in young (3-month-old), adult (12-month-old) and aged (24-month-old) male Sprague-Dawley rats. Activation of D-1 receptors with DA, apomorphine or fenoldopam enhanced accumulation of cAMP in the frontal cortex of young rats. The stimulatory effect elicited by DA on cAMP generation declined by about 20% in adult rats. No further decline in cAMP accumulation was noticeable in aged animals. The response to dopaminergic agonists was blocked by the D-1 receptor antagonist SCH 29390 in the three age groups examined. The presence of D-2 receptors, negatively coupled to cAMP generation, was demonstrated by incubating frontal cortex membrane particles with SCH 23390 and then with DA. This inhibitory response, was also elicited with D-2 receptor agonists quinpirole or bromocriptine in the absence of SCH 23390 in which these compounds produced a decrease in cAMP. The decrease in cAMP caused following D-2 receptor stimulation was shown to be enhanced with age. No difference was observed between the three age groups of animals in the activation of cAMP production by forskolin. The present data suggest a selective decrease in the coupling between the D-1 receptor and cAMP generation in the frontal cortex of adult and aged rats and of an age-dependent increase in the coupling between the D-2 receptor and cAMP inhibition. The functional consequences of these biochemical changes may have important implications in the aging of the rat frontal cortex.     

Dissociation between spontaneously emitted and apomorphine-induced stereotypy in Peromyscus maniculatus bairdii

Stereotyped behavior is repetitive, topographically invariant motor activity that lacks an obvious function. We have previously characterized the spontaneous and persistent stereotypies that occur in deer mice housed in standard laboratory cages. Providing these animals with enriched environments markedly reduces their vulnerability to develop stereotypic behavioral repertoires, thus enabling us to generate behaviorally distinct (stereotypic and nonstereotypic) mice of the same species. As stereotypic behaviors are readily induced by systemic administration of a dopamine (DA) agonist, the present study tested whether apomorphine would induce stereotypies in environmentally enriched (nonstereotypic) deer mice that were topographically similar to the stereotypies that are spontaneously emitted by standard-caged (stereotypic) deer mice. The effects of apomorphine were also evaluated in the standard-caged (stereotypic) deer mice. DA agonist-induced behaviors in nonstereotypic mice included stereotypies that were largely topographically distinct from spontaneously emitted stereotypies; apomorphine failed to produce statistically significant elevations in two of the three stereotypic behaviors typical of standard-caged deer mice. Furthermore, there was no evidence of increased DA receptor sensitivity in stereotypic mice. Thus, environmentally related stereotypy is distinct from systemically administered DA agonist-induced stereotypy, and is not exacerbated by such drug treatment. The results obtained do provide support, however, for a limited involvement of the DA system in the mediation of these behaviors.     

Behavioral and neurochemical modifications induced by apomorphine treatment

By means of quantitative receptor autoradiography, the modifications of dopamine (DA) receptors in selected target regions of A₉ and A₁₀ DA neurons were studied after chronic apomorphine treatment, at a dosage able to induce behavioral changes that are claimed to be due to the activation of the target areas of A₉ neurons. An increase in [³H]spiperone binding sites (cold (+)-butaclamol) was observed in the dorsal and ventrolateral striatum, receiving fibers from A₉ neurons, while there were no changes in the ventromedial and ventrocentral striatum, in the nucleus accumbens and in the cerebral cortex receiving fibers from A₁₀ neurons. Our results suggest that the anatomical division of the DA target regions corresponds with a functional one.     

Bicuculline administration into ventromedial hypothalamus: effects on fear and regional brain monoamines and GABA concentrations in rats

The effects of bicuculline methiodide administration into ventromedial hypothalamus (15 ng per site, bilaterally) on fear behavior and monoamines (NA, DA, 5-HT) and GABA in structures of the brain defensive system (hypothalamus, midbrain gray matter, amygdala, hippocampus and frontal cortex) were studied. Fear behavior was examined in the modified version of light-dark transition test. The time out from the illuminated compartment of chamber, the time spent there and number of returns to the illuminated compartment was measured. Additionally motor activity, i.e., number of crossings and rearings in dark as well as in the illuminated part of compartment, was registered. Blockade of GABAA receptors in the ventromedial hypothalamus resulted in increased fear behavior, i.e. decrease of time out from illuminated compartment and decrease of the time spent there. Motor behavior remained unchanged. HPLC analysis showed reduction of GABA concentration in all investigated brain structures. An increase of NA concentration in all examined structures with exception of the hypothalamus without effect on MHPG/NA was observed as well. Dopamine level remained unchanged, but DOPAC/DA ratio increased in all structures, except frontal cortex. Also HVA/DA ratio increased in the hypothalamus and midbrain. 5-HT concentration increased only in midbrain, 5-HIAA increased in midbrain and in frontal cortex, and 5-HIAA/5-HT ratio increased only in frontal cortex. These results indicate that GABA-ergic and monoaminergic systems remain in functional interactions and that these interactions may play an important role in the neurochemical regulation of fear behavior. The possible mechanism of GABA--monoaminergic interactions is discussed.     

Hypertension-induced changes in monoamine receptors in the prefrontal cortex of rhesus monkeys

Hypertension affects approximately 60 million people in the United States. Recent studies have demonstrated that hypertension may produce progressive changes in the CNS. The present study is focused on reports in the literature that hypertension may significantly alter neurotransmitter systems, particularly dopamine (DA) and norepinephrine (NE). To address this, DA and norepinephrine (NE) receptor binding was assessed in the prefrontal cortex (PFC) of 15 male rhesus monkeys using on-the-slide in vitro assays for the DA1, NE alpha1 and NE alpha2 receptors as well as for the DA and NE uptake transporters. Eight monkeys underwent surgical coarctation of the mid-thoracic aorta which produced sustained, untreated hypertension as defined by a systolic pressure above 150 mm Hg. Compared with normotensive controls, chronic, untreated hypertension produced a significant decrease in DA1 and NE alpha1 receptor binding and an increase in DA uptake (DAU) receptor binding in the prefrontal cortex. While the mechanisms by which untreated hypertension alters DA and NE receptors is not known, the use of this non-human primate model should provide the means to uncover neurobiological changes that occur with untreated hypertension.     

Chronic but not acute clomipramine alters the effect of NMDA receptor regulation of dopamine release in rat frontal cortex

The effect of acute or chronic treatment with the antidepressant clomipramine (CIM) on N-methyl-D-aspartate (NMDA) evoked release of dopamine (DA) in the frontal cortex of the rat has been studied using microdialysis. Acute injection of CIM (10 or 20 mg/kg) caused a decrease in dialysate DA in the frontal cortex. Infusion of 25-100 microM NMDA into the frontal cortex decreased DA release in this region. When NMDA infusion was preceded by a single injection of CIM no marked differences between NMDA and NMDA + CIM treated groups were observed. Chronic (15 day) treatment with CIM (10 or 20 mg/kg) caused a dose-dependent increase in basal extracellular DA. In these animals, however, the effects of infusion of NMDA on DA release in the cortex were greatly attenuated or abolished. This suggests that adaptive changes occur in NMDA receptor function during treatment with an antidepressant. The possible significance of this in the aetiology and treatment of depression is discussed.     

D-1 dopamine receptors labelled with 3H-SCH 23390: decrease in the striatum of aged rats

The effects of age on the binding parameters of 3H-SCH 23390, the most selective D-1 dopamine receptor ligand available at present, were studied in membrane preparations from rat striatum. When compared with 3 month old animals, there was a significant decrease in the density of 3H-SCH 23390 binding sites in 20 month old (-37%) and in 26 month old (-44%) rats, without alterations in the apparent dissociation constant values. No significant changes in the density or affinity of D-1 DA receptors were observed in 14 month old rats. In view of the behavioural effects mediated by D-1 DA receptors recently described (i.e., grooming, stereotypies and EEG desynchronization), the decrease in the density of these receptors in senescent rats may underlie some of the age-related alterations in dopaminergic functions in the rat brain.     

Evidence for involvement of N-methyl-D-aspartate receptor in tonic inhibitory control of dopaminergic transmission in rat medial frontal cortex

Bilateral infusion of DL-2-amino-5-phosphonovalerate (DL-APV) (which is a competitive antagonist for N-methyl-D-aspartate (NMDA) receptor) into the medial frontal cortex of conscious rats increased the amount of 3,4-dihydroxyphenylacetic acid (DOPAC) and the DOPAC/dopamine (DA) ratio in the cortical area. Moreover, intra-prefrontal injection of DL-APV, D-APV, DL-2-amino-7-phosphonoheptanoate and 3-[(+/-])-2-carboxypiperazin-4-yl]-propyl-1-phosphonate (which are selective NMDA receptor antagonists), but not the L-isomer of APV and gamma-glutamyl-aminomethyl sulphonate (a relative antagonist for non-NMDA receptors), facilitated prefrontal DA utilization in a NMDA-reversible manner. These findings suggest that NMDA-type excitatory amino acid receptors may be involved in a tonic inhibitory regulation of dopaminergic transmission in the medial frontal cortex in vivo.     

The influence of selective serotonin reuptake inhibitors (SSRIs) on the pharmacokinetics of thioridazine and its metabolites: in vivo and in vitro studies.

Due to its psychotropic profile, thioridazine is a neuroleptic suitable for a combination with antidepressants in a number of complex psychiatric illnesses. However, because of its serious side-effects, such a combination with selective serotonin reuptake inhibitors (SSRIs) which inhibit cytochrome P-450 may be dangerous. The aim of the present study was to investigate a possible impact of SSRIs on the pharmacokinetics and metabolism of thioridazine in a steady state in rats. Thioridazine (10 mg/kg) was injected intraperitoneally, twice a day, for two weeks, alone or jointly with one of the antidepressants (fluoxetine, fluvoxamine or sertraline). Concentrations of thioridazine and its main metabolites (2-sulfoxide = mesoridazine; 2-sulfone = sulforidazine; 5-sulfoxide = ring sulfoxide and N-desmethylthiorid-azine) were assessed in the blood plasma and brain at 30 min, 6 and 12 h after the last dose of the drugs using an HPLC method. Fluoxetine potently increased (up to 13 times!) the concentrations of thioridazine and its metabolites in the plasma, especially after 6 and 12 h. Moreover, an increase in the sum of concentrations of tioridazine + metabolites and thioridazine/metabolite ratios was observed. In vitro studies with control liver microsomes, as well as with microsomes of rats treated chronically with fluoxetine show that the changes in the thioridazine pharmacokinetics may be attributed to the competitive (N-demethylation, Ki = 23 microM) and mixed inhibition (2- and 5-sulfoxidation, Ki = 60 microM and 34 microM, respectively) of thioridazine metabolism by fluoxetine, and to the adaptive changes produced by chronic administration of fluoxetine, as reflected by inhibition of N-demethylation and formation of sulforidazine. Sertraline seemed to have a tendency to decrease thioridazine concentration in vivo, though in vitro studies showed that - like fluoxetine - it competitively or via mixed mechanism inhibited the three metabolic pathways of thioridazine (Ki = 41 microM, 64 microM and 47 microM, respectively). Chronic treatment with sertraline stimulated thioridazine 2- and 5-sulfoxidation, which may be responsible for the observed tendency of sertraline to decrease concentrations of the neuroleptic. In the case of fluvoxamine, a tendency to increase the thioridazine level was observed, which may be connected with the competitive or mixed inhibition of thioridazine N-demethylation and 2-sulfoxidation by the antidepressant (Ki = 17 microM and 167 microM, respectively). Repeated administration of fluvoxamine did not produce any changes in the activity of thioridazine-metabolizing enzymes. In conclusion, of the SSRIs studied, only fluoxetine produces a substantial increase in the thioridazine level in the plasma and brain. In the case of fluvoxamine, a tendency to increase the thioridazine level should be considered. Coadministration of thioridazine and sertraline seems to be safe, though a tendency to decrease the thioridazine level may be expected.     

Cold restraint alters dopamine metabolism in frontal cortex, nucleus accumbens and neostriatum

The concentrations of dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) were assayed in the striatum, nucleus accumbens and frontal cortex of rats following 2 hours of cold restraint. The concentration of DA was significantly decreased in both the striatum (-16%) and nucleus accumbens (-41%) relative to unstressed controls. The content of DOPAC was significantly increased in both striatum (+56%) and frontal cortex (+76%), but not in nucleus accumbens. The DOPAC/DA ratio was increased in all three regions, that in frontal cortex approaching three-fold. These results extend earlier findings of an activation by acute stressors of frontal cortex DA metabolism, but suggest an involvement of other DA systems as well. The finding of the greatest response in frontal cortex, and the previous observations that this was the only region to show significant changes, may be ascribed to the suggested lack of presynaptic autoreceptors in this region.     

Gangliosides increase the survival of lesioned nigral dopamine neurons and favour the recovery of dopaminergic synaptic function in striatum of rats by collateral sprouting*

The effects of chronic ganglioside treatment GM-1 (10 mg/kg, i. p., once daily for 56 days) have been evaluated on the degenerative and regenerative features of nigrostriatal dopamine (DA) neurons following a partial lesion by tyrosine hydroxylase immunocytochemistry in combination with morphometrical analysis and by quantitative DA receptor autoradiography. Chronic GM-1 treatment resulted in the maintenance in the number of DA cell bodies, terminals and striatal area on the lesioned side and also increased dendrite length of the DA nerve cells in the zona reticulata on that side. The lesion induced DA receptor supersensitivity was counteracted by chronic treatment with GM-1 and the apomorphine induced rotational behaviour was significantly reduced. The hypothesis is introduced that following ganglioside treatment some lesioned DA nerve cells do not degenerate, but elongate their dendrites to give increased trophic support to DA cell bodies with intact DA axons. These increased dendro-dendritic interactions may enable the unlesioned DA cells to increase the density of their striatal nerve terminal networks via collateral sprouting leading to recovery of dopaminergic synaptic function as evidenced in the receptor autoradiographical and behavioural analysis. Gangliosides may therefore possibly represent a new type of drug in the treatment of Parkinson's disease and aging processes in DA systems.     

Different effects of acute and chronic stress on two dopamine-mediated behaviors in the mouse

After two hours of immobilization stress, C57BL/6 mice presented a significant reduction of spontaneous locomotion and a slight reduction of spontaneous climbing. The effect of stress on locomotor activity disappeared after ten daily sessions of immobilization while this chronic treatment increased the effect of stress on spontaneous climbing. Twenty-four hr after the last stressful experience the mice showed an increase of spontaneous locomotion and a decrease of spontaneous climbing in comparison with unstressed mice. Following a single exposure to immobilization stress, an increase of DOPAC/DA and HVA/DA ratios was found in the striatum and in the nucleus accumbens. These effects were still evident following repeated exposure to this stressor but disappeared 24 hr after the last of ten daily stressful experiences. Finally, chronically stressed mice, tested 24 hr after the last stressful experience, showed an increased sensitivity to the inhibitory effects of low doses of apomorphine on climbing behavior and a decreased sensitivity to the inhibitory effects of the same doses of the dopamine agonist on locomotion. These results are discussed in terms of altered sensitivity of different populations of dopamine receptors following chronic stress.     

Effects on brain cholecystokinin-8-sulfate and serotonin in adult rats after neonatal reduction of catecholamines

Depletion of dopamine (DA) in neonatal rats by cerebroventricular infusion of 6-hydroxydopamine with desmethylimipramine resulted in a significant decrease in mesolimbic immunoreactive cholecystokinin-8-sulfate (CCK-8SI) and a marked increase in striatal serotonin in 10-week-old rats. In contrast, neonatal norepinephrine (NE) depletion did not affect either CCK-8SI or serotonin in the frontal cortex, anterior cingulate cortex or hippocampus. Neonatal DA and NE depletion did not induce an increase in striatal serotonin.     

Homeostatic regulation of excitatory synapses on striatal medium spiny neurons expressing the D2 dopamine receptor

Striatal medium spiny neurons (MSNs) are contacted by glutamatergic axon terminals originating from cortex, thalamus and other regions. The striatum is also innervated by dopaminergic (DAergic) terminals, some of which release glutamate as a co-transmitter. Despite evidence for functional DA release at birth in the striatum, the role of DA in the establishment of striatal circuitry is unclear. In light of recent work suggesting activity-dependent homeostatic regulation of glutamatergic terminals on MSNs expressing the D2 DA receptor (D2-MSNs), we used primary co-cultures to test the hypothesis that stimulation of DA and glutamate receptors regulates the homeostasis of glutamatergic synapses on MSNs. Co-culture of D2-MSNs with mesencephalic DA neurons or with cortical neurons produced an increase in spines and functional glutamate synapses expressing VGLUT2 or VGLUT1, respectively. The density of VGLUT2-positive terminals was reduced by the conditional knockout of this gene from DA neurons. In the presence of both mesencephalic and cortical neurons, the density of synapses reached the same total, compatible with the possibility of a homeostatic mechanism capping excitatory synaptic density. Blockade of D2 receptors increased the density of cortical and mesencephalic glutamatergic terminals, without changing MSN spine density or mEPSC frequency. Combined blockade of AMPA and NMDA glutamate receptors increased the density of cortical terminals and decreased that of mesencephalic VGLUT2-positive terminals, with no net change in total excitatory terminal density or in mEPSC frequency. These results suggest that DA and glutamate signaling regulate excitatory inputs to striatal D2-MSNs at both the pre- and postsynaptic level, under the influence of a homeostatic mechanism controlling functional output of the circuit.     

Behavioral and neurochemical alterations evoked by p-Chlorophenylalanine application in rats examined in the light-dark crossing test

The aim of the present study is to examine the effects of serotonin synthesis inhibition with p-Chlorophenylalanine (p-CPA) in rats on (1) anxiety behavior examined in the light-dark crossing test and, (2) regional brain concentration of monoamines (NA, DA and 5-HT) and their metabolites (MHPG, DOPAC, HVA and 5-HIAA) as well as GABA in the hypothalamus, amygdala, hippocampus, midbrain central gray matter and the frontal cortex. Treatment of animals with p-CPA produced a significant increase in time out from the illuminated part of the chamber and in time of locomotor activity in the illuminated part of the chamber. HPLC analysis showed a significant reduction of 5-HT and 5-HIAA concentration in all examined brain regions with the exception of the frontal cortex. Additionally, a significant decrease in DA and its metabolites, DOPAC and HVA occurred in the hypothalamus and amygdala. Moreover, we observed a significant decrease in frontal cortex NA concentration after p-CPA administration. The results of our study suggest that administration of p-CPA is effective in reduction of anxiety through depletion of 5-HT accompanied by diminution of catecholamines, especially DA and its metabolites in the main emotional brain regions.     

S-adenosyl-L-methionine prevents 5-HT(1A) receptors up-regulation induced by acute imipramine in the frontal cortex of the rat

S-adenosyl-L-methionine (SAM) has shown efficacy in speeding the onset of the antidepressant effect of imipramine in depressed patients. This effect may be related to their interactions at the serotonin(1A) (5-HT(1A)) receptors. Acute imipramine up-regulated the frontal cortex 5-HT(1A) receptors (B(max), 51.5 +/- 8.4 fmol/mg protein) vs. saline (B(max), 27.5 +/- 5.9 fmol/mg protein), and did not show antidepressant effect. Acute SAM and imipramine+SAM did not modify frontal cortex 5-HT(1A) receptors, and showed antidepressant effects (decrease of the immobility response of 26%, P<0.01; and 47%, P<0.001) vs. saline. All the chronic treatments showed antidepressant effects and up-regulated the hippocampus 5-HT(1A) receptors. SAM prevents the 5-HT(1A) receptor up-regulation induced by acute imipramine in the frontal cortex. This mechanism may contribute to imipramine's antidepressant effect.     

Effects of single and continuous administration of amyloid beta-peptide (25-35) on adenylyl cyclase activity and the somatostatinergic system in the rat frontal and parietal cortex

It is unknown whether the amyloid beta-peptide (Abeta), a principal component found in extracellular neuritic plaques in the brain of patients with Alzheimer's disease (AD), is capable of altering adenylyl cyclase (AC) activity and the somatostatin (SRIF) receptor-effector system in the cerebral cortex of the patients. Therefore, the objective of this study was to investigate the effect of the beta fragment, beta (25-35), on AC activity and the somatostatinergic system in the rat frontoparietal cortex. A single dose of beta (25-35) (10microg) injected intracerebroventricularly significantly decreased the density of SRIF receptors (27.4%) and increased their affinity (32.2%) in the frontoparietal cortex. The inhibitory effect of SRIF on basal and forskolin (FK)-stimulated AC activity was significantly lower in the beta (25-35)-treated rats when compared with controls. beta (25-35) did not modify Gialpha1, Gialpha2 nor Gialpha3 levels in membranes from the frontoparietal cortex. Continuous infusion of the peptide induced a decrease in the SRIF receptor density in this brain area to a similar extent as that observed 14 days after the single administration of the peptide. Likewise, this treatment decreased the SRIF receptor density in the frontal cortex (15.3%) and parietal cortex (27.2%). This effect was accompanied by a decrease in the SRIF-mediated inhibition of FK-stimulated AC activity (from 41.6% to 25.6%) in the frontal cortex as well by a decrease in basal AC activity (from 36.9% to 31.6%) and FK-stimulated AC activity (from 35.6% to 27.1%) in the parietal cortex. Continuous infusion of Abeta (25-35) had no effect on Gialpha1, Gialpha2 or Gialpha3 levels in membranes from frontal and parietal cortex. However, this treatment caused a decrease in SRIF-like immunoreactivity content in the parietal (38.9%) and frontal (20.4%) cortex. These results suggest that Abeta might be involved in the alterations of somatostatinergic system reported in AD.     

Phencyclidine increases extracellular dopamine metabolites in rat medial frontal cortex as measured by in vivo dialysis

An acute intraperitoneal injection of phencyclidine (PCP) caused a tetrodotoxin-reversible increase in extracellular release of 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the dialysates from the medial frontal cortex of the rat. Moreover, there was an increase in the tissue content of DOPAC and HVA with acceleration of dopamine (DA), but not noradrenaline, utilization in the cortical area after systemic administration of PCP. These results suggest that PCP facilitates DA metabolism in the medial frontal cortex by increasing impulse flow in the DA neurons projecting to the prefrontal region.     

Functional in vivo interaction between growth hormone and dopamine systems are correlated to changes in striatal somatostatin levels as detected by voltammetry

The effects of growth hormone (GH) and somatomedin C (SmC), as well as those of apomorphine, dopamine (DA) agonist, or haloperidol (DA antagonist), upon the size of striatal voltammetric peaks 2 and 5 were investigated. Local intrastriatal injections of GH or SmC were followed by an increase in the height of both peak 2 (corresponding to the oxidation of extracellular dihydro-phenylacetic acid, DOPAC, a metabolite of DA) and peak 5 (which may represent the oxidation of striatal extracellular somatostatin, SRIF). Treatment with haloperidol also increased the size of the striatal catechol peak but was responsible for a reduction of the neuropeptidergic signal. By contrast, apomorphine determined a decrease in striatal peak 2 (DOPAC) while increasing the levels of peak 5 (SRIF). The data further support the chemical identification of peak 5 at +800 mV as related to the in vivo oxidation of SRIF; in addition they indicate the presence of a functional relationship between this neuropeptide and the GH and DA systems in the striatum of anaesthetised rats.     

Blockade of M1 muscarinic acetylcholine receptors modulates the methamphetamine-induced psychomotor stimulant effect

Muscarinic acetylcholine receptors (M1-M5) regulate many key functions of the CNS and peripheral nervous system. In the present study, the role of M1 muscarinic receptors (M1R) in the psychomotor stimulant and sensitizing properties of methamphetamine (METH) is investigated using molecular, neurochemical, and behavioral approaches. Acute and repeated treatment with METH increased M1R mRNA expression in the frontal cortex and the CA2 region of the hippocampus. Repeated treatment with METH also increased M1R mRNA expression in the dentate gyrus. Dicyclomine, an M1R antagonist, did not affect the psychomotor effect of METH, but it attenuated METH-induced increases in the dopamine (DA) efflux in the nucleus accumbens (NAc). Dicyclomine enhanced the psychomotor effect of METH after repeated treatment with METH and 8.0 mg/kg of dicyclomine, and also augmented the increase in the NAc DA overflow evoked by repeated METH treatment. These results suggest that M1R plays a role in the METH-induced psychomotor stimulant effect by changing the release of DA in the NAc of mice.