Fear memory-induced alterations in the mRNA expression of G proteins in the mouse brain and the impact of immediate posttraining treatment with morphine.

Disturbances in fear-evoked signal transduction in the hippocampus (HP), the nuclei of the amygdala (AMY), and the prefrontal cortex (PFC) underlie anxiety-related disorders. However, the molecular mechanisms underlying these effects remain elusive. Heterotrimeric G proteins (GPs) are divided into the following four families based on the intracellular activity of their alpha subunit (Gα): Gα(s) proteins stimulate cyclic AMP (cAMP) generation, Gα(i/o) proteins inhibit the cAMP pathway, Gα(q/11) proteins increase the intracellular Ca++ concentration and the inositol trisphosphate level, and Gα(12/13) proteins activate monomeric GP-Rho. In the present study, we assessed the effects of a fear memory procedure on the mRNA expression of the Gα subunits of all four GP families in the HP, AMY and PFC. C57BL/6 J mice were subjected to a fear conditioning (FC) procedure followed by a contextual or cued fear memory test (CTX-R and CS-R, respectively). Morphine (MOR, 1 mg/kg/ip) was injected immediately after FC to prevent the fear consolidation process. Real-time quantitative PCR was used to measure the mRNA expression levels of Gα subunits at 1 h after FC, 24 h after FC, and 1 h after the CTX-R or CS-R. In the HP, the mRNA levels of Gα(s), Gα(12) and Gα(11) were higher at 1 h after training. Gα(s) levels were slightly lower when consolidation was stabilized and after the CS-R. The mRNA levels of Gα(12) were increased at 1 h after FC, returned to control levels at 24 h after FC and increased again with the CTX-R. The increase in the Gα(11) level persisted at 24 h after FC and after CTX-R. In the AMY, no specific changes were induced by FC. In the PFC, CTX-R was accompanied by a decrease in Gα(i/o) mRNA levels; however, only Gα(i2) downregulation was prevented by MOR treatment. Hence, the FC-evoked changes in Gα mRNA expression were observed mainly in the HP and connected primarily to contextual learning. These results suggest that the activation of signaling pathways by Gα(s) and Gα(12) is required to begin the fear memory consolidation process in the HP, while signal transduction via Gα(11) is implicated in the maintenance of fear consolidation. In the PFC, the downregulation of Gα(i2) appears to be related to the contextual learning of fear.

Brief maternal separation affects brain α1-adrenoceptors and apoptotic signaling in adult mice.

Exposure to adversity during early life is a risk factor for the development of different mood and psychiatric disorders, including depressive-like behaviors. Here, neonatal mice were temporarily but repeatedly (day 1 to day 13) separated from mothers and placed in a testing environment containing a layer of odorless clean bedding (CB). We assessed in adult animals the impact of this early experience on binding sites and mRNA expression of α1-adrenergic receptor subtypes, heat shock proteins (HSPs) and proapoptotic and antiapoptotic members of the Bcl-2 family proteins in different brain regions involved in processing of olfactory information and rewarding stimuli. We found that repeated exposure to CB experience produced anhedonic-like behavior in terms of reduced saccharin intake and α1-adrenoceptor downregulation in piriform and somatosensory cortices, hippocampus, amygdala and discrete thalamic nuclei. We also found a selective decrease of α1B-adrenoceptor binding sites in the cingulate cortex and hippocampus and an increase of hippocampal α1A and α1B receptor, but not of α1D-adrenoceptor, mRNA levels. Moreover, while a significant decrease of antiapoptotic heat shock proteins Hsp72 and Hsp90 was identified in the prefrontal cortex, a parallel increase of antiapoptotic members of Bcl-2 family proteins was found at the hippocampal level. Together, these data provide evidence that the early exposure to CB experience produced enduring downregulation of α1-adrenoceptors in the prefrontal-limbic forebrain/limbic midbrain network, which plays a key role in the processing of olfactory information and reaction to rewarding stimuli. Finally, these data show that CB experience can "prime" the hippocampal circuitry and promote the expression of antiapoptotic factors that can confer potential neuroprotection to subsequent adversity.

[Neurochemistry of impulsiveness and aggression]. / Neurochemia impulsywnosci i agresji.

Aggression is the most frequent social reaction among animals and men, and plays an important role in survival of the fittest. The change of social conditions in the course of development of human civilisation rendered some forms of aggression counter-adaptive, but the neurobiological mechanism of expression of aggression have not fundamentally changed in the last stages of human evolution. The two different kinds of aggression: emotional, serving mainly as a threat, and rational, predatory, serving for the attainment of goal in the most effective way, have different anatomical and neurobiological background and reciprocally inhibit each other. Aggression is modulated by several neurotransmitter and hormonal systems, of which the key role is seemingly played by testosterone, a hormone involved in domination behaviour, and serotonin, whose deficit results in increased impulsiveness.

Early maternal separation: a rodent model of depression and a prevailing human condition.

The early life of most mammals is spent in close contact with the mother, and for the neonate, early maternal separation is a traumatic event that, depending on various conditions, may shape its behavioral and neurochemical phenotype in adulthood. Studies on rodents demonstrated that a very brief separation followed by increased maternal care may positively affect the development of the offspring but that prolonged separation causes significant amounts of stress. The consequences of this stress (particularly the hyperreactivity of the HPA (hypothalamic-pituitary-adrenal) axis are expressed in adulthood and persist for life. Maternal separation in rodents, particularly rats, was used as a model for various psychotic conditions, especially depression. The most popular separation procedure of a 3-h daily separation from the second to the 12th postpartum day yields a depression model of high construct and predictive validity. The results of studies on maternal separation in rats and monkeys prompt a discussion of the consequences of traditional procedures in the maternity wards of developed countries where attention is focused on the hygiene of the neonates and not on their psychological needs. This alternate focus results in a drastic limitation of mother-infant contact and prolonged periods of separation. It is tempting to speculate that differences in the course and severity of various mental disorders, which are usually less prevalent in underdeveloped countries than in developed countries (as noted by Kraepelin), may be related to different modes of infant care. Only recently has so-called kangaroo mother care (establishing mother-infant skin-to-skin contact immediately after birth) become popular in developed countries. In addition to its instant benefits for the neonates, this procedure may also be beneficial for the mental health of the offspring in adulthood.

Morphine-induced place preference affects mRNA expression of G protein α subunits in rat brain.

BACKGROUND: The conditioned place preference (CPP) test is an animal model serving to assess addictive potential of drugs in which environmental cues become associated with the subjective effects of drugs of abuse. Morphine, a known addictive drug, is an agonist of opioid receptors that couple to the G(i/o) family of guanine nucleotide-binding proteins (GP). We have recently found that chronic treatment with morphine affects mRNA levels of GPs that are not coupled to opioid receptors (OR). Therefore, in this study, we investigated the influence of morphine-induced CPP on mRNA expression of the Gα subunits, G(i/o), G(s), G(q/11), and G(12), in the rat prefrontal cortex (PFC) and nucleus accumbens (NAc) using standard PCR techniques. METHODS: CPP and NO-CPP experiments were conducted; Wistar rats were either subjected to the standard CPP procedure or were injected with morphine (or saline) in their home cage. All rats were decapitated 24 h after the last injection. RESULTS: We found that mRNA levels of Gα(q), Gα(11) and Gα(12) were increased after morphine in non-conditioned treatment in the PFC but remained unchanged in the NAc. In rats showing conditioned place preference to morphine, levels of Gα(i2) in the PFC and levels of Gα(oA) in the NAc were diminished by ≈58% and ≈30%, respectively (p < 0.05 vs. saline), but levels of Gα(s-l) in NAc were increased (≈60%, p = 0.05). CONCLUSION: Our data indicate that only G(i/o) and G(s) were specifically changed in animals after morphine-induced CPP, thus suggesting that the effect was related to learning environmental cues associated with morphine.

Changes induced by formalin pain in central alpha1-adrenoceptor density are modulated by adenosine receptor agonists.

We aimed to elucidate the role of alpha(1)-adrenoceptors in adenosine analgesia in the formalin test. Formalin was injected into the hind paw of male CD-1 mice after injection of adenosine A(1) or A(2a) receptor agonists, CPA, [N(6)-cyclopentyladenosine], and CGS21680 [2-p-(2-carboxyethyl)-phenylethylamino-5'-N-ethylcarboxamidoadenosine hydrochloride]. In the behavioral experiment, alpha(1)-adrenoceptors were blocked by an alpha(1)-adrenoceptor antagonist prazosin, 0.01 mg/kg i.p., and the time mice spent paw licking was recorded for the early (0-15 min) and late (15-60 min) phase of formalin pain. In the neurochemical experiments, mice were killed 15 or 45 min after formalin injection. The density of alpha(1)-adrenoceptors was assessed in various brain areas and in the lumbar spinal cord by [(3)H]prazosin autoradiography. Adenosine agonists produced analgesia in both phases of formalin pain, while prazosin showed a tendency to pronociceptive action in the late phase, and antagonized the effect of CGS21680. After formalin injection, alpha(1)-adrenoceptor density was elevated in some brain areas, mainly in the late phase (some contralateral amygdaloid and ipsilateral thalamic nuclei) and depressed in others (early phase in the ipsilateral spinal cord and late phase in both ipsi- and contralateral sensorimotor cortex). Elevation of alpha(1)-adrenoceptor density, which may be interpreted as a defensive response, did not develop in several cases of CPA-pretreated mice. This suggests that the analgesic effect of adenosine A(1) receptor activation renders the defensive response unnecessary. The depression of alpha(1)-adrenoceptors may suggest development of hypersensitivity in a given structure, and this was antagonized by CGS21680, suggesting the role of A(2a) receptors in control of inflammatory formalin pain.

Paroxetine pretreatment does not change the effects induced in the rat cortical beta-adrenergic receptor system by repetitive transcranial magnetic stimulation and electroconvulsive shock.

Repetitive transcranial magnetic stimulation (rTMS) has been proposed as a clinically effective antidepressant treatment, but meta-analysis suggests that its efficacy is marginal. We investigated whether the administration of rTMS together with paroxetine would enhance its effects on the beta-adrenergic system of the rat. We compared our results with the effects of electroconvulsive shock therapy (ECS). The experiment was performed for 12 d on male Wistar rats that received a physical treatment of either rTMS (B=1.4 T, f=10 Hz, 300 s) or ECS (I=130 mA, f=50 Hz, t=500 ms), preceded by sterile water or paroxetine (10 mg/kg i.p. 30 min earlier). All rats were decapitated 24 h after the final treatment. Cyclic AMP (cAMP) was measured in cortical slices prelabelled with [3H]adenine and stimulated with noradrenaline. beta-adrenoceptor parameters (Bmax and KD) were assessed in the P2 fraction of cortical homogenates using [3H]CGP 12177 as a ligand. ECS resulted in down-regulation of both the cAMP response and beta-adrenoceptor density, while rTMS depressed only the responsiveness of the cAMP-generating system. Paroxetine, which was only effective in dampening the cAMP response, did not change the effects of either physical treatment. The data suggest that any possible interaction between paroxetine and rTMS or ECS does not involve the beta-adrenergic mechanisms.

3-Methoxytyramine, an extraneuronal dopamine metabolite plays a physiological role in the brain as an inhibitory regulator of catecholaminergic activity.

3-Methoxytyramine (3-MT), an extraneuronal metabolite of dopamine, present in the synaptic cleft at a very low amount (low nanomolar range), comparable to dopamine concentration, is generally regarded as a biologically inactive compound. We have shown in this study that 3-MT binds to the rat noradrenergic cortical alpha(1) and striatal dopamine D(1) and D(2) receptors in nanomolar concentration range, and to cortical alpha(2) adrenoceptor at low micromolar concentration. Bilateral intrastriatal injections of 3-MT (0.25 micromol in 0.5 microl) did not affect significantly locomotor activity in naive rats but strongly antagonized amphetamine-induced (1 mg/kg s.c.) hypermotility. Biochemical studies in rat brain structures showed that 3-MT behaved as an antagonist of the noradrenergic system, i.e. accelerated noradrenaline metabolism and counteracted the inhibitory effect of amphetamine on the rate of noradrenaline metabolism. In contrast to a general view about the lack of physiological role of monoamine metabolites, these results for the first time strongly suggest that an extraneuronal metabolite of dopamine, 3-MT plays an important physiological role as an inhibitory regulator counteracting excessive stimulation of catecholaminergic neurons in the striatum.

Does the presence of morphine counteract adaptive changes in expression of G-protein alpha subunits mRNA induced by chronic morphine treatment?

Opiate dependence develops due to changes in intracellular signaling caused by long-term exposure to morphine. Here we investigated changes in the mRNA expression of the main classes of G-protein alpha (Galpha) subunits in various brain regions in morphine-dependent rats. Rats received increasing doses of morphine, 10-50 mg/kg, b.i.d., for 14 days. G-protein alpha-subunit mRNA expression was determined shortly following the conclusion of chronic morphine administration (2 h after the final dose) and during withdrawal (48 h after the final dose). Significant changes in mRNA expression for Galpha subunits were observed in several brain areas during withdrawal, while the changes were much less evident or absent 2 h after the final drug injection. Changes in mRNA expression were particularly evident in the nucleus accumbens (increases in Galpha(12), Galpha(q), Galpha(11), and Galpha(o) during withdrawal, increase in Galpha(i) and decrease in Galpha(s) just following treatment). The direction of the changes, which were not all significant, for Galpha(12), Galpha(q), and Galpha(11) was generally consistent in the amygdala and prefrontal cortex; changes in G proteins coupled to the adenylyl cyclase cascade were less consistent. These results suggest that morphine dependence leads to alterations in intracellular signaling, which are reflected in changes in the expression of genes encoding various G proteins. The results may explain why signs of opiate dependence are not expressed during chronic administration of morphine, but only after cessation of the treatment.

Diverse effects of GABA-mimetic drugs on cocaine-evoked self-administration and discriminative stimulus effects in rats.

RATIONALE: Recent data indicate that gamma-aminobutyric acid (GABA) is a modulator of behavioral responses to cocaine. OBJECTIVE: The efficacy of gabapentin (a cyclic GABA analogue), tiagabine (a GABA reuptake inhibitor), or vigabatrin (an inhibitor of GABA transaminase and reuptake) to alter cocaine-seeking behavior and discriminative effects was examined in rats. MATERIALS AND METHODS: Rats were trained to press a lever for cocaine (0.5 mg/kg per infusion) paired with a cue (light + tone) using a fixed ratio (FR) 5 schedule of reinforcement. After extinction, the cocaine-seeking behavior was reinstated by cocaine priming (10 mg/kg). Another group of rats was trained to discriminate cocaine (10 mg/kg) from saline in a two-lever FR 20 task. RESULTS: Vigabatrin (150-250 mg/kg) decreased cocaine-maintained responding, whereas tiagabine (10 mg/kg) significantly reduced responses on the "active" lever. Vigabatrin (150-250 mg/kg) significantly decreased responding to the cocaine-priming dose and a nonsignificant attenuation of cocaine-induced reinstatement was seen after tiagabine (5-10 mg/kg). Gabapentin (10-30 mg/kg) failed to alter maintenance of cocaine self-administration or drug-induced reinstatement. Pretreatment with either gabapentin, tiagabine, or vigabatrin resulted in neither reinstatement of cocaine seeking nor alterations in cocaine discrimination. CONCLUSIONS: Our study demonstrates that vigabatrin (only at the 150 mg/kg dose) exerted inhibitory actions on cocaine-maintained responding and attenuated the reinstatement of extinguishing responding more effectively than gabapentin or tiagabine and with less evidence of motor impairment than the latter drugs. Present findings do not support a role for gabapentin or tiagabine for the possible treatment of cocaine relapse, whereas albeit limited effects of vigabatrin may be seen.

Various GABA-mimetic drugs differently affect cocaine-evoked hyperlocomotion and sensitization.

To substantiate the notion that cocaine behavioral effects may be influenced by gamma-aminobutyric acid (GABA) neurotransmission male Wistar rats were injected with gabapentin (a cyclic GABA analogue), tiagabine (a GABA reuptake inhibitor), or vigabatrin (a GABA transaminase inhibitor) before acute or repeated treatment with cocaine evoking either locomotor hyperactivation or sensitization. Gabapentin (1-30 mg/kg), tiagabine (2.5-10 mg/kg) or vigabatrin (75-250 mg/kg) attenuated the cocaine (10 mg/kg)-induced hyperactivation and in the highest doses they also decreased basal locomotor activation. Vigabatrin (75-250 mg/kg) dose-dependently reduced the development of cocaine sensitization in rats treated repeatedly (days 1-5) with cocaine (10 mg/kg) and then challenged with cocaine (10 mg/kg) following 5-day withdrawal; the remaining drugs were ineffective. When injected acutely with a cocaine challenge dose, gabapentin (3-10 mg/kg) or vigabatrin (150 mg/kg), but not tiagabine, significantly attenuated the expression of cocaine sensitization. The present results show that enhanced GABA-ergic neurotransmission exerted inhibitory actions on acute responses to cocaine, however, only in a case of vigabatrin the inhibition seems to be unrelated to the inhibitory effect of the drugs on basal locomotor activity. The finding that vigabatrin protected against the development and the expression of cocaine sensitization further supports its therapeutic potential in the treatment of cocaine dependence.

The mechanism of 1,2,3,4-tetrahydroisoquinolines neuroprotection: the importance of free radicals scavenging properties and inhibition of glutamate-induced excitotoxicity.

1-Methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), unlike several other tetrahydroisoquinolines, displays neuroprotective properties. To elucidate this action we compared the effects of 1MeTIQ with 1,2,3,4-tetrahydroisoquinoline (TIQ), a compound sharing many activities with 1MeTIQ (among them reducing free radicals formed during dopamine catabolism), but offering no clear neuroprotection. We found that the compounds similarly inhibit free-radical generation in an abiotic system, as well as indices of neurotoxicity (caspase-3 activity and lactate dehydrogenase release) induced by glutamate in mouse embryonic primary cell cultures (a preparation resistant to NMDA toxicity). However, in granular cell cultures obtained from 7-day-old rats, 1MeTIQ prevented the glutamate-induced cell death and 45Ca2+ influx, whereas TIQ did not. This suggested a specific action of 1MeTIQ on NMDA receptors, which was confirmed by the inhibition of [3H]MK-801 binding by 1MeTIQ. Finally, we demonstrated in an in vivo microdialysis experiment that 1MeTIQ prevents kainate-induced release of excitatory amino acids from the rat frontal cortex. Our results indicate that 1MeTIQ, in contrast to TIQ, offers a unique and complex mechanism of neuroprotection in which antagonism to the glutamatergic system may play a very important role. The results suggest the potential of 1MeTIQ as a therapeutic agent in various neurodegenarative illnesses of the central nervous system.

Conditioned rewarding stimulus associated with cocaine self-administration reverses the depression of catecholamine brain systems following cocaine withdrawal in rats.

Craving phenomena are related to induction of substance-seeking behaviour by stimuli associated with the availability of the drug. We investigated the changes in monoamine metabolism in regions of the brains of rats that, following a period of training of cocaine self-administration, were either killed 2 h after the last session or underwent extinction trials, during which cocaine was withdrawn. During the training, acoustic and visual stimuli announced the availability of cocaine. After 10 d of daily extinction trials, rats were re-introduced into the cage, and a signal associated with cocaine availability was applied to half of the animals. The rats were immediately killed and concentrations of dopamine and serotonin and their metabolites in various brain areas, and the concentration of noradrenaline and MHPG in the brainstem were assessed to calculate their metabolism rate indices. In rats self-administering cocaine, the levels of metabolites of all three amines were depressed, indicating a depression of the activity of monoaminergic systems. In the period of extinction, the dopamine levels in the nucleus accumbens and striatum and the level of the noradrenaline metabolite, MHPG, in the brainstem were reduced, suggesting a long-lasting disturbance of the catecholaminergic system, while serotonin levels and metabolism returned to normal values. The presence of the signal associated with previous cocaine availability, which invariably caused the reinstatement of cocaine-seeking behaviour annulled the changes observed in the group receiving no stimulus, bringing the concentration values of dopamine, and dopamine and noradrenaline metabolites to yoked-saline control rats. The results suggest that the stabilized self-administration of cocaine depresses the activity of all biogenic amine systems, and the changes in serotonin system are reversible, in contrast to those observed in catecholaminergic systems, which show the signs of a long-lasting impairment. The stimulus associated with cocaine availability activates the catecholaminergic system in animals after extinction procedure.

Nicotine potentiates imipramine-induced effects on catecholamine metabolism: possible relation to antidepressant activity.

Following our behavioral studies demonstrating augmentation of imipramine action by concomitant administration of nicotine, we investigated the effects of one or 14 days of treatment (twice daily) with imipramine and nicotine on dopamine metabolism in various brain areas of rat and noradrenaline in the brain stem. In addition, we evaluated the responses of this metabolism to apomorphine challenge in the rat. Generally, chronic treatment of imipramine and nicotine produced opposite effects to acute administration. As revealed by HPLC, dopamine metabolism in the nucleus accumbens was slightly decreased after 14 days of treatment with imipramine, and co-administration of nicotine resulted in a significant and much more pronounced depression of dopamine metabolism in all investigated dopaminergic structures. Such biochemical effects suggested the development of a compensatory mechanism related with hypersensitivity of dopamine D(2) receptors in the mesolimbic and nigrostriatal system. Chronic administration of imipramine produced an opposite effect to the acute one in the brain stem noradrenergic system, like it was observed in dopaminergic structures. Significant inhibition of noradrenaline metabolism after acute administration of imipramine may be explained by its inhibitory effect on noradrenaline reuptake process. In contrast, chronic imipramine administration had no effect on noradrenaline metabolism what indicated the development of subsensitivity of (2)-adrenoceptors in the brain stem responsible for the rate of noradrenaline metabolism. Apomorphine alone decreased metabolism of both catecholamine, dopamine and noradrenaline through activation of dopamine D(2) receptors which are located also on noradrenergic neurons. The biochemical response to apomorphine in terms of dopamine metabolism was not changed by chronic administration of the investigated drugs but noradrenaline metabolism in the brain stem was strongly attenuated after a combined treatment of imipramine and nicotine. The present data demonstrate facilitation and potentiation of biochemical antidepressant-like effects of imipramine by nicotine co-treatment. We suggest that nicotine may potentiate the antidepressant-like effects of imipramine by promoting some plastic changes in the brain within dopamine and noradrenaline system considerably more strongly than imipramine alone.

Effect of cocaine sensitization on alpha1-adrenoceptors in brain regions of the rat: an autoradiographic analysis.

We investigated the effects of repeated intermittent cocaine treatment, resulting in behavioral sensitization, on the density of alpha(1)-adrenoceptors in the rat brain measured by quantitative in vitro autoradiography of [(3)H]prazosin. Animals were decapitated following a short (2 h) and long (48 h) withdrawal period after an injection of cocaine (10 mg/kg) on day 10 given to either cocaine-naive (saline daily, days 1-5) or cocaine-sensitized (cocaine 10 mg/kg daily, days 1-5) rats. In cocaine-naive rats, significant decreases in alpha(1)-adrenoceptors 2 h after a single dose of cocaine were observed in the amygdaloid nuclei and hippocampus; the decreases in the centromedial nucleus of the amygdala persisted until 48th hour of withdrawal. On the contrary, increases in alpha(1)-adrenoceptors after 2-h withdrawal were seen in the nucleus accumbens core and retrosplenial cortex. In cocaine-sensitized rats, the density of alpha(1)-adrenoceptors 2 h after the challenge with cocaine increased in the centrolateral amygdala, while in the granular retrosplenial cortex and in the most of thalamic nuclei, the densities of alpha(1)-adrenoceptors decreased. After 48-h withdrawal, the density of alpha(1)-adrenoceptors increased in the nucleus accumbens core and shell (by 21% and 58%, respectively), and in the amygdaloid centromedial and basolateral nuclei (by ca. 24%), while the decline was still observed in some thalamic nuclei. Our study shows for the first time that cocaine sensitization produces significant (dependent on the withdrawal time) alterations in the alpha(1)-adrenoceptor density, and the changes in some parts of the thalamus seem to be related to processes of cocaine relapses.

Nicotine produces antidepressant-like actions: Behavioral and neurochemical evidence.

Converging lines of evidence indicate the involvement of nicotinic acetylcholine receptors in depressive illness and antidepressant drug action. We investigated the effects of sub-chronic and chronic treatment with imipramine, nicotine and their combination on: (a) the ability of a dopamine-mimetic challenge to produce locomotor stimulation and (b) cortical density of beta-adrenoceptors. One week of treatment with imipramine (10 mg/kg, twice daily) did not result in an altered response to the apomorphine (0.15 mg/kg) challenge, but after 2 weeks, the imipramine-treated rats demonstrated hyperactivity. Conversely, such increased locomotor response was observed in rats treated with nicotine (0.4 mg/kg, twice daily) for 1 but not for 2 weeks. Groups treated with nicotine+imipramine for 1 and 2 weeks demonstrated equally high hyperactivity in response to the apomorphine challenge. This effect was not different from the effects of 1-week treatment with nicotine or 2-week treatment with imipramine. The density of beta-adrenoceptors was equally decreased by 2 (but not 1) weeks of the treatment with imipramine, nicotine and their combination. The present behavioral and neurochemical data suggest the antidepressant-like effect of the chronic treatment with nicotine. It appears that the potentiation of the dopamine-mimetic-induced hyperactivity cannot be explained by beta-adrenoceptor down-regulation.

Carane derivative stereoisomers of different local anaesthetic and antiplatelet activity similarly potentiate forskolin-stimulated cyclic AMP response and bind to beta-adrenoceptors in the rat brain cortex.

A carane derivative, KP-23 [RS](-)-4-(2-hydroxy-3)N-isopropylamino)-propoxyimino)-cis-carane, was earlier described as a potential local anaesthetic and antiplatelet agent, and the following studies revealed that its R and S stereoisomers, KP-23R and KP-23S, have different potencies in the infiltration anaesthesia and platelet aggregation tests. The effects of these stereoisomers on the cyclic AMP (cAMP) generating system and the displacement of [(3)H]CGP 12177 (a beta-adrenoceptor ligand) from its binding sites in the rat cerebral cortical tissue were investigated. The stereoisomers did not affect the basal cAMP level, but, at concentrations between 10(-4) and 10(-3) M, they elevated the forskolin-induced accumulation of cAMP with similar potency. The compounds displaced [(3)H]CGP 12177, however the stereoisomer R was less potent than the racemic KP-23 and the S form (K(i) = 64.1 +/- 5.9 nM, 161.1 +/- 10 nM and 62.1 +/- 5.6 nM for KP-23, KP-23R and KP-23S, respectively). The fact that the stereoisomers differed in both tests only slightly, if at all, suggests that their pharmacological effects are not related to the action on the beta-adrenoceptor/adenylate cyclase system.

Protective effect of 1-methyl-1,2,3,4-tetrahydroisoquinoline against dopaminergic neurodegeneration in the extrapyramidal structures produced by intracerebral injection of rotenone.

The aim of this paper was to investigate whether rotenone, a pesticide causing experimental parkinsonism, causes direct damage to dopaminergic structure when injected intracerebrally and whether this action may be prevented by peripheral administration of 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), an endogenous compound with anti-dopaminergic activity. Male Wistar rats were injected unilaterally into the median forebrain bundle with 2 microg rotenone, and received 1MeTIQ, 50 mg/kg i.p. 1 h before and then daily for 21 d. To compare the effect of intracerebral and peripheral treatment, rotenone was also given once or for 7 d in a dose of 10 mg/kg s.c. Dopamine, serotonin and their metabolites were assessed by HPLC in the substantia nigra and striatum. While a single subcutaneous rotenone dose did not produce any change in striatal dopamine metabolism, the multiple treatments resulted in changes suggesting a shift in the metabolism towards oxidative desamination and reduction of O-methylation. In contrast to systemic injections, intracerebral-administered rotenone produced a decrease in dopamine and its metabolites content in the striatum (dopamine decrease by 70%) and substantia nigra (dopamine decrease by 35%), without affecting the serotonin system. As those changes were observed 21 d after the injection of rotenone, they suggest a durable neurotoxic effect. The treatment with 1MeTIQ strongly reduced the fall of striatal dopamine concentration. The data suggest that rotenone given peripherally affects metabolic processes in dopaminergic neurons, and this seems to result from its neurotoxic action, which may be observed after an intracerebral injection. 1MeTIQ is able to counteract the damaging action of rotenone and seems to be a potential neuroprotective agent.

A possible physiological role for cerebral tetrahydroisoquinolines.

Tetrahydroisoquinolines present in the mammalian brain, 1,2,3,4-tetrahydroisoquinoline (TIQ) and salsolinol, suspected to cause neurodegeneration leading to Parkinson's disease, were investigated to find their possible physiological role. To this aim their behavioral and receptor effects induced after a single dose were tested in mice and rats. Both compounds do not affect significantly the basal locomotor activity, very effectively block hyperactivity induced by apomorphine (rats) and amphetamine (mice), only partially block hyperactivity induced by scopolamine, do not affect locomotor stimulation induced by cocaine, and strongly augment the running fit induced by morphine (mice). They do not produce extrapyramidal symptoms and do not potentiate haloperidol-induced catalepsy (rats). TIQ and salsolinol do not displace antagonists of several receptors (including D(1) and D(2)) from their binding sites, but displace the agonists of Alpha(2)-adrenoceptors, [(3)H]clonidine and of dopamine receptors, [(3)H]apomorphine. The results indicate that salsolinol and TIQ act as specific antagonists of agonistic conformation of dopamine receptors, and owing to that may play a role of endogenous feed-back regulators of the dopaminergic system. Those properties make tetrahydroisoquinolines potential antidopaminergic drugs devoid of extrapyramidal effects, with possible application in substance addiction disorder as anti-craving agents.

1-methyl-1,2,3,4-tetrahydroisoquinoline protects against rotenone-induced mortality and biochemical changes in rat brain.

The effect of single and multiple administration of the neurotoxic pesticide, rotenone, and the potentially neuroprotective compound, 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), on the concentration of dopamine and its metabolites (homovanillic acid-HVA, 3,4-dihydroxyphenylacetic acid-DOPAC, and 3-methoxytyramine-3-MT)) in three brain areas was studied by high-performance liquid chromatography (HPLC) with electrochemical detection in Wistar rats. The rate of dopamine catabolism in the striatum along the N-oxidative and O-methylation pathways was assessed by calculation of the ratio of dopamine metabolites to dopamine. In addition, the effect of rotenone on mortality and general behavior of rats was investigated. We have found that the neurotoxic pesticide, rotenone, administered in a single dose (12 mg/kg s.c.) did not produce evident behavioral or biochemical effects. In contrast, repeated administration of rotenone in doses (12-15 mg/kg) causing abnormalities in general behavior, produced considerable mortality and dramatic increases in dopamine metabolism, which may be ascribed to an increase in the oxidative pathway. Interestingly, it depressed the concentration of the extracellular dopamine metabolite, 3-MT. These behavioral and biochemical changes were effectively counteracted by administration of 1MeTIQ before each dose of rotenone. In summary, we demonstrated that multiple systemic rotenone injections are strongly toxic, and induce alterations of cerebral dopamine metabolism, and that 1MeTIQ may be considered as a potential protective agent against environmental factors affecting the function of the dopaminergic system.