Compulsive sexual activity induced by p-chlorophenylalanine in normal and pinealectomized male rats.

p-Chlorophenylalanine depletes brain serotonin and induces longlasting sexual excitation in male rats. The effect of p-chlorophenylalanine is potentiated by pargyline. Administration of 5-hydroxytryptophan to rats treated with p-chlorophenylalanine plus pargyline blocks the sexual excitation. p-Chlorophenylalanine also elicits sexual excitation in pinealectomized rats; this effect is not mediated by the lack of indole hormones in the pineal but may be the consequence of depletion of 5-hydroxytryptophan in the brain and the resulting imbalance between 5-hydroxytryptophan and catecholamine activity in the central nervous system.

Efficacy of an Hypericum perforatum (St. John's wort) extract in preventing and reverting a condition of escape deficit in rats.

The treatment of unselected depressed patients with an hydro-alcoholic extract of Hypericum perforatum has been reported to have an efficacy similar to that of classical antidepressants. In the present report, the effects of H. perforatum were studied on three animal models of depression: (i) an acute form of escape deficit (ED) induced by an unavoidable stress; (ii) a chronic model of ED, which can be maintained by the administration of mild stressors on alternate days; (iii) a model of anhedonia based on the finding that repeated stressors prevent the development of an appetitive behavior induced by vanilla sugar in satiated rats fed ad libitum. H. perforatum acutely protected animals from the sequelae of unavoidable stress; such an effect was partially prevented by the administration of SCH 23390 or (-)-pindolol. Moreover, H. perforatum reverted the ED maintained by repeated stressors and preserved the animal's capacity to learn to operate for earning a positive reinforcer. It was concluded that H. perforatum contains some active principle(s) endowed with antidepressant activity.

Dizocilpine infusion has a different effect in the development of morphine and cocaine sensitization: behavioral and neurochemical aspects.

The stimulation of glutamate receptors plays a relevant role in the development of behavioral sensitization to psychostimulants, while less clear results have been obtained on their role in morphine sensitization. We addressed this issue by comparing the development of cocaine and morphine sensitization under a continuous s.c. infusion of the N-methyl-D-aspartate (NMDA) antagonist dizocilpine (0.1 mg/kg/24 h). Moreover, we studied the expression of NMDA and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor subunits in discrete limbic areas of rats sensitized to morphine or cocaine with or without the concomitant dizocilpine infusion. It was observed that dizocilpine infusion did not prevent the development of morphine sensitization, while it prevented the development of tolerance to morphine-induced analgesia. Finally, morphine-sensitized animals did not present any modification in the subunit expression of glutamate receptors in the brain areas examined. In agreement with previous results, we found that dizocilpine infusion prevented the development of cocaine sensitization. Moreover, we observed that rats sensitized to cocaine presented a significant increase in the levels of GLUR1, NR1 and NR2B, in the nucleus accumbens, and of NR2B in the hippocampus compared to control animals. Such modifications were absent in rats administered cocaine under dizocilpine infusion. We conclude that: (i) morphine sensitization is a neuroadaptive phenomenon which does not appear to require NMDA receptor activity in order to develop; (ii) cocaine sensitization is clearly dependent on NMDA receptor activity, as dizocilpine infusion prevented the occurrence of glutamate receptors modifications as well as the development of sensitization.

Acquisition of an appetitive behavior reverses the effects of long-term treatment with lithium in rats.

Rats exposed to a long-term treatment with lithium chloride develop a deficit of avoidance accompanied by a reduction in the basal levels of extraneuronal dopamine and in dopamine accumulation in the nucleus accumbens shell after acute uptake inhibition. Such a condition is similar to that of an experimental model of depression induced by exposing rats to a chronic stress procedure. Rats exposed to chronic stress are also unable to acquire an appetitive behavior sustained by a highly palatable food. Thus, it was studied whether rats fed a diet containing lithium would develop an appetitive behavior induced by a pure hedonic stimulus. Rats on the lithium diet developed a clear-cut escape deficit condition accompanied by a decreased dopamine output in the nucleus accumbens shell; nevertheless, they learned the appetitive behavior within a similar period to controls. The development of the appetitive behavior coincided with the recovery of the capacity to avoid a noxious stimulus and with the return of the dopaminergic transmission in the nucleus accumbens shell to values similar to those of control rats. It may be concluded that the mechanism of action underlying the behavioral and neurochemical sequelae of a chronic stress is distinct from that of the analogous effects produced by lithium.

Acquisition of a palatable-food-sustained appetitive behavior in satiated rats is dependent on the dopaminergic response to this food in limbic areas.

Rats exposed to repeated unavoidable stress show decreased dopamine output in the nucleus accumbens shell (NAcS) and do not acquire vanilla sugar (VS)-sustained appetitive behavior (VAB). Rats treated with lithium for 3 weeks also show decreased NAcS dopamine output, yet they acquire VAB. Feeding a novel palatable food increases extraneuronal dopamine levels in the NAcS and medial prefrontal cortex (mPFC) in rats. In order to investigate the role of food-induced dopamine release in VAB acquisition, we studied by microdialysis the dopaminergic response in the NAcS and mPFC to the presentation and consumption of VS in satiated control rats, and in satiated rats exposed to repeated stress or lithium treatment. The dopaminergic response to VS was also studied in rats familiar with VS, or that had acquired VAB. In control rats, VS feeding was accompanied by increased dopamine output in the NAcS and mPFC, and one-trial habituation to this effect developed in the NAcS. Rats exposed to a 7-day stress showed reduced interest in VS pellets, and when fed VS they did not show a dopaminergic response in the NAcS and mPFC. Lithium-treated rats rapidly ate VS pellets and showed increased dopamine output in the NAcS and mPFC, with no habituation in the NAcS response. Rats familiar with VS and rats that had already learned VAB ate VS pellets. The first group showed a lower dopaminergic response to VS consumption than the control group, but the latter showed no dopaminergic response in the NAcS and mPFC. We propose that the limbic dopaminergic response to a novel palatable food plays a role in associative learning and that it is predictive of the competence to learn an appetitive behavior. Moreover, in our experimental conditions a phasic increase in mesolimbic dopamine no longer signals the VS stimulus once it has become a reinforcer in an appetitive task.

NMDA receptors play an anti-aggregating role in human platelets.

Receptors for different monoamines, peptides and other neurohormones are present in the plasma membrane of platelets, and the sophisticated process of haemostasis is regulated by the interplay of their physiologic agonists (1). The recent report of a platelet binding site for phencyclidine (2) suggested a possible role of N-methyl-D-aspartate (NMDA) receptors in platelet function. Isotherms of [3H]-glutamate (GLU), [3H]-CGP-39653, [3H]-glycine (GLY) and [3H]-MK-801 carried out in platelet membranes yielded Bmax and Kd values for these ligands similar to those present in neurons, and NMDA only partially displaced [3H]-GLU. In neurons [3H]-MK-801 binding is potentiated by GLU and/or GLY and, being specific for the open NMDA receptor channel species, it has a functional meaning. In platelet membranes neither GLU and/or GLY increased [3H]-MK-801 binding; thus suggesting that NMDA receptors in platelets are different from those present in neurons. GLU or NMDA alone did not induce platelet aggregation. However, both amino acids were antagonistic on the aggregating activity of arachidonic acid (AA), NMDA being 3 orders of magnitude more active than GLU, and NMDA also antagonized adenosine diphosphate (ADP) and platelet aggregating factor (PAF) induced platelet aggregation. Finally, NMDA increased cAMP levels in intact platelets, and such an effect did not occur in a Ca(2+)-free medium; yet, cAMP increase was not antagonized by the calmodulin inhibitor trifluoperazine (TFP). It was concluded that platelet membranes carry an NMDA receptor, functionally distinct from the neuronal one, which seems to play an anti-aggregating role.

Lack of copulatory behaviour in male castrated rats after p-chlorophenylalanine.

1 The effect of p-chlorophenylalanine (PCPA) on the copulatory behaviour of normal and castrated male rats with females in oestrus was studied.2 Castration 2 months before the experiment completely prevented the increased copulatory behaviour produced by PCPA in normal rats.3 The administration of testosterone restored the copulatory behaviour in the castrated rats indicating that testosterone is essential for this behaviour.

Further insights into the anti-aggregating activity of NMDA in human platelets.

1. In the present study the effect of N-methyl-D-aspartate (NMDA) on thromboxane B2 synthesis and on [Ca2+]i was studied in human platelets. 2. NMDA (10(-7) M) completely inhibited the synthesis of thromboxane B2 from exogenous arachidonic acid (AA), while it did not interfere with the aggregating effect of the thromboxane A2 receptor agonist U-46619. 3. NMDA (0.1 microM - 10 microM) dose-dependently increased intracellular calcium in washed platelets preloaded with fura 2 AM, and this effect was not additive with that of AA. 4. NMDA shifted the dose-response curve of AA to the right. At the highest AA concentrations platelet aggregation was not inhibited. 5. The antiaggregating effect of NMDA was not antagonized by N(G)-monomethyl-L-arginine (L-NMMA), a nitric oxide synthase (NOS) inhibitor. 6. Finally, NMDA (0.01 nM - 100 nM) associated with either aspirin or indomethacin significantly potentiated the antiaggregating activity of both cyclo-oxygenase inhibitors. 7. It was concluded that NMDA is a potent inhibitor of platelet aggregation and thromboxane B2 synthesis in human platelet rich plasma (PRP).

Failure by tricyclic antidepressants to affect the increase of dopamine extracellular concentrations produced by haloperidol in the caudate and accumbens nuclei of rats.

The effect of different inhibitors of monoamine uptake on dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) neuronal outflow was studied in the caudate and accumbens nuclei of rats, using the in vivo brain microdialysis method coupled to HPLC electrochemical detection. Under conditions of DA receptor blockade (as produced by the i.p. administration of 0.25 mg/kg of haloperidol), cocaine, GBR-12909 and d-amphetamine increased the concentration of extracellular DA beyond the effect produced by haloperidol alone in both areas studied. GBR-12909 and cocaine also increased DOPAC concentration, while d-amphetamine decreased it. On the contrary, the tricyclic antidepressants (TCA), desipramine and chloripramine, failed to modify the effect of haloperidol on DA and DOPAC neuronal outflow. It was concluded that: (a) nonadrenergic and serotonergic nerve terminals do not take up DA released from dopaminergic neurons, and (b) TCA have no effect on dopaminergic terminals.

Endogenous codeine and morphine are stored in specific brain neurons.

Codeine and morphine have been detected in mammalian brain by radioimmunoassay (RIA), and in brain and other tissues by gas-chromatography/mass-spectrometry (GCMS) in different laboratories. It has been also shown that rat liver can synthesize the skeleton of the morphine molecule, thus suggesting that this alkaloid, which is the prototype of mu-receptor agonists, plays a physiological role in brain. We report the presence of morphine-like immunoreactive compounds inside the cell body, fibers and terminals of neurons in different brain areas. Moreover, neurons localized in the same brain areas were capable of accumulating and storing [3H]morphine slowly infused intracerebroventricularly (i.c.v.) through an osmotic minipump.

Rats sensitized to morphine are resistant to the behavioral effects of an unavoidable stress.

In agreement with the results of other authors, rats sensitized to morphine and challenged with 5 mg/kg of morphine after 7 days of wash-out showed intense stereotyped movements, the expression of which was selectively antagonized by SCH 23390. Sensitized rats were exposed to an unavoidable stress (which consistently produces an escape deficit in control animals) after 3, 7 and 21 days of morphine wash-out. Twenty-four hours after the unavoidable stress, animals were tested for their capacity to escape and their performance was compared to that of control-stressed and naive rats. Morphine sensitization completely prevented the development of escape deficit. This protective effect was similar to that induced by a chronic imipramine treatment and, like the effect of imipramine, it was antagonized by the administration of SCH 23390 before the unavoidable stress. However, it was not affected by the administration of naloxone. Moreover, when rats presenting a clear-cut escape deficit, induced by a 10-day treatment with SKF 38393, were exposed to the morphine sensitization protocol, a complete recovery of their capacity to avoid a noxious stimulus was observed. Finally, the down-regulation of both the number of D(1)-dopamine receptors and of the coupled adenylyl cyclase activity in the pre-frontal cortex (PFC) produced by long-term SKF 38393 administration was reverted by the superimposed morphine sensitization. Thus, the condition of morphine sensitization appears to share several common effects with chronic imipramine treatment.

Dopamine output in the nucleus accumbens shell is related to the acquisition and the retention of a motivated appetitive behavior in rats.

Chronic stress exposure consistently impairs the reactivity to aversive and pleasurable stimuli in rats; these behavioral modifications are associated with a decrease in dopamine output in the nucleus accumbens shell (NAcS). However, when rats that have already acquired an appetitive behavior are exposed to chronic stress, they develop an impaired reactivity to avoidable aversive stimuli while retaining the appetitive behavior. The dissociation between these two behavioral traits was used to study whether the decreased dopaminergic activity in the NAcS was connected to either of the two deficits. Dopamine output was studied through microdialysis as dopamine accumulation following re-uptake inhibition by cocaine. When rats that had previously acquired the appetitive behavior were exposed to chronic stress, they showed a dopaminergic transmission in the NAcS similar to that of controls and significantly higher than that of chronically stressed animals. Thus, dopamine output in the NAcS was consistently associated to the acquisition and maintenance of appetitive behavior, while the expression of a deficit in avoidance appeared to be independent of it.

Long-term lithium administration abolishes the resistance to stress in rats sensitized to morphine.

Morphine sensitized rats appear protected from the sequelae of an unavoidable stress: when exposed to stress (after a 7-day morphine wash-out) and then tested for escape, they perform like naive animals. This protection appears similar to that induced by chronic imipramine treatment, as it is antagonized by the inhibition of D(1)-dopamine receptors before exposure to unavoidable stress. Repeated unavoidable stress induces in rats a condition characterized by hyporeactivity to noxious stimuli and reverted by long-term antidepressant treatments, and this state is regarded as an experimental model of depression. The resistance to stress in morphine sensitized rats could be considered as the behavioral counterpart of the sensitivity to stress in control rats, i.e. as a model of mania. The aim of the present study was to validate such a putative model by studying whether the resistance to stress induced by morphine sensitization would respond to a long-term administration of lithium, the reference antimanic drug. Long-term lithium treatment induces in rats a condition of hyporeactivity to noxious stimuli, accompanied by decreased levels of dopamine in the nucleus accumbens shell. In morphine sensitized rats chronic lithium abolished the resistance to stress, but it did not modify the D(1)-dopamine receptor mediated response to morphine, nor did it modify the levels of extraneuronal dopamine in the nucleus accumbens shell. Thus, lithium treatment abolished the resistance to stress in morphine sensitized rats, conferring predictive validity to the paradigm. Moreover, it did so through a mechanism which appeared to be independent of D(1)-dopamine receptor activity.

Long-term imipramine effects are prevented by NMDA receptor blockade.

Long-term exposure to different antidepressant treatments induces increased motor response to central stimulants, due to a selective supersensitivity of dopamine D2 receptors in the limbic areas. Such an effect is accompanied by down-regulation of dopamine D1 receptor number, and by a decreased response of adenylyl cyclase to dopamine stimulation in the limbic system. Moreover, the number of beta-adrenergic receptors and the response of adenylyl cyclase to beta-adrenergic stimulation in the cortex result to be reduced. The present data confirms that imipramine (10 mg/kg twice a day for 3 weeks) produces such effects, and shows that the co-administration of imipramine with MK-801 (administered by a subcutaneously implanted osmotic minipump delivering 0.05 mg/kg/day of the compound) prevented the occurrence of both the behavioral supersensitivity to quinpirole, and the decrease of dopamine D1 and beta-adrenergic receptor function.

The effects of long-term administration of rubidium or lithium on reactivity to stress and on dopamine output in the nucleus accumbens in rats.

Rubidium and lithium are alkali metals belonging to the same periodic series as sodium, potassium and cesium. In the present report the effects of lithium and rubidium on animal reactivity to stressful stimuli and on dopamine output in the nucleus accumbens were studied. A dose-response curve with rubidium, administered acutely before exposure to unavoidable stress, showed a maximal protective activity on escape deficit development at the dose of 0. 41 mEq/kg. Rubidium injected at doses of 0.008-0.08 mEq/kg 72 h before the unavoidable stress had the same efficacy as the acute 0. 41 mEq/kg dose. Tolerance to the effect of rubidium developed after 9 days of treatment and, on day 15, rats presented a spontaneous escape deficit. The acute effect of lithium, administered for 3.5 days at the dose of 0.8 mEq/kg, i.p. twice a day before the exposure to unavoidable stress, was analogous to that of rubidium, but after repeated treatment a spontaneous escape deficit developed. Rats showing an escape deficit secondary to chronic stress also presented decreased extraneuronal dopamine concentrations in the nucleus accumbens. Accordingly, microdialysis studies showed significantly lower extracellular dopamine levels in rats chronically treated with lithium or rubidium compared to control animals. Cocaine (5 mg/kg i. p.) administered acutely increased extracellular dopamine concentrations in control rats, as well as in rats chronically stressed or chronically treated with lithium or rubidium. However, the dopamine increase was significantly higher in controls compared to the other groups. In conclusion, long-term treatment with lithium or rubidium, or the exposure to chronic stress, produced a condition of behavioral hypo-reactivity accompanied by a decreased dopamine output in the nucleus accumbens.

Selective adenylate cyclase increase in the limbic area of long-term imipramine-treated rats.

Long-term administration of imipramine to rats produced an increase in the Vmax of forskolin- or guanylylimidodiphosphate (Gpp(NH))p-activated adenylate cyclase only in the limbic area. This effect was prevented by the daily administration of alpha-methyl-p-tyrosine (alpha-MPT), given together with imipramine, at a dose (50 mg/kg) which had no effect on adenylate cyclase activity per se. The time course of the effects of chronic imipramine on dopaminergic transmission in the limbic area showed that the decrease in both D-1 receptor number and adenylate cyclase stimulation by dopamine (DA) reached significance on day 8 of treatment and were maximal on day 15. The Vmax of the enzyme started to increase on day 15 and was further increased on day 21. Possible mechanisms underlying these effects are discussed.

Failure of morphine to increase striatal 3,4-dihydroxyphenylacetic acid in fasted rats.

Morphine given to rats fed ad libitum increased the striatal levels of DA and DOPAC while naloxone had no effect. Conversely, after prolonged fasting, morphine failed to increase both DA and DOPAC, while naloxone markedly decreased striatal DOPAC concentration. Intermediate sensitivity to the DOPAC increasing effect of morphine and to the DOPAC decreasing effect of naloxone was present in rats trained to eat their daily meal within 2 h, at different times after feeding. It is concluded that striatal DA synthesis in fasted rats is sustained by an increased endorphin-like activity.

Intranigral kainic acid: evidence for nigral non-dopaminergic neurons controlling posture and behavior in a manner opposite to the dopaminergic ones.

The unilateral, intranigral administration of kainic acid (k.a.) produced a syndrome characterized by early sequelae of contra- and ipsilateral circling and by a chronic contralateral turning associated with moderate loss of neurons in the pars reticulata. The acute contralateral circling seems to be related to dopaminergic nigro-neostriatal neuron stimulation, since it was prevented by previous intranigral injections of 6-OHDA. The acute ipsilateral circling and the chronic contralateral turning, on the other hand, seem to be independent of the integrity of the dopaminergic system and may be due to an initial stimulation, followed by destruction, of a nigral neuronal system which mediates turning behavior in a manner opposite to that of nigro-striatal dopamine. Treatment with D-amphetamine or apomorphine changed the contralateral into ipsilateral turning, while haloperidol potentiated the contralateral turning. Bilateral injection of k.a. into the nigra resulted in chronic stereotyped sniffing and gnawing, which were not inhibited by haloperidol. Moreover, haloperidol did not produce catalepsy in these animals. It is suggested that the intranigral k.a. injection destroyed a neuronal system antagonistic to dopamine and resulted in a reduction of the response to DA-receptor stimulation of the c. striatum.

In utero exposure to methadone produces a stable decrease of the cortex 5-HT transport system in rats.

Female rats were made dependent to high doses of morphine (400 mg/kg per day) or methadone (60-80 mg/kg per day) and subsequently exposed to adult males. None of the morphine-dependent rats became pregnant while there were no differences in the gestation time and number of young in the litter born to each rat in the methadone and the control groups. The values for muscarinic, serotonergic and opiate receptors measured in the whole brain of the offspring at 1 and 7 days after birth were similar in the control and in the methadone-exposed animals. Brain beta-receptors were lower in rats exposed in utero to methadone at 1 and 7 days. [3H]IMI binding sites were steadily and significantly lower in the whole brain of rats born to methadone-dependent animals than in the brain of controls. In 2 month old animals, the decrease of [3H]IMI binding capacity was associated with a decrease of [3H]5HT uptake. In the light of these findings the authors propose a unifying hypothesis to explain the altered reactivity to morphine of animals treated with opiates during gestation.