Effects of clonidine and some α-adrenoreceptor blocking agents on avoidance conditioned reflexes in rats: Their interactions and antagonism by atropine

Clonidine (150 g/kg s. c.) depressed avoidance conditioned reflexes in Long-Evans rats. Some -adrenoceptor blocking agents piperoxan, tolazoline, phentolamine, dibenamine and phenoxybenzamine also depressed these reflexes. Yohimbine (1–2 mg/kg) was found to have no significant effect. Atropine (10 to 15 mg/kg i. p.) antagonized the depression of conditioned avoidance reflexes induced by -adrenoceptor blocking agents as well as the depression produced by clonidine.Yohimbine (1–2 mg/kg) reduced the depressing effects of clonidine on avoidance conditioned reflexes. In rats treated with piperoxan (10 mg/kg) the effects of clonidine were reduced. In these rats the depression of conditioned reflexes was less than in rats treated with clonidine or piperoxan alone.These experiments suggest that clonidine depressed avoidance conditioned reflexes in rats by activating central -adrenoceptors. It is proposed that -adrenoceptor blocking agents might act as partial agonists.     

Involvement of alpha 2-adrenoceptors in the activation of kininogenesis by clonidine

The effects of clonidine and alpha 2-adrenoceptor blocking agents, yohimbine and tolazoline on kininogen and prekallikrein levels in plasma and on blood pressure were investigated in male Wistar rats. Clonidine (0.5 mg/kg) decreased concentration of kininogen. Yohimbine (3 mg/kg) and tolazoline (5 mg/kg) counteracted this effect of clonidine and increased the kininogen level in plasma. Clonidine did not change the level of prekallikrein, however in rats receiving alpha 2-adrenoceptor blockers alone or together with clonidine kallikrein utilization was reduced, which indicated indirectly that the level of prekallikrein in plasma augmented. Yohimbine and tolazoline in the same doses (3 mg/kg and 5 mg/kg resp.) increased the blood pressure and counteracted the hypotensive action of clonidine, which indicated the presynaptic alpha 2-adrenoceptor blocking effect of the doses used. The data indicate that clonidine-induced activation of kininogenesis as well as its hypotensive effect are counteracted by yohimbine and tolazoline in doses blocking specifically presynaptic alpha 2-adrenoceptors. It can be suggested that alpha 2-adrenoceptors are involved in the kininogenesis-activating effect of clonidine.     

Cognitive enhancers prevent the hypoxia-induced disruption of conditioned avoidance response

Six cognitive enhancer compounds, the new class of central nervous system (CNS)-active drugs (vinpocetine, vincamine, dihydroergotoxine, nicergoline, piracetam, and meclofenoxate) were tested before acquired two-way active avoidance response of spontaneously hypertensive (SH) rats had been disrupted by lowered environmental oxygen concentration (6% oxygen). Exposure to normobaric hypoxia reduced the number of conditioned avoidance responses (CAR) by 44%; at the same time the spontaneous locomotor activity of the animals (measured by the alteration in the number of intertrial crossings) was not considerably decreased, and latency times of conditioned responses were not lengthened significantly. The compounds tested showed protective effect against hypoxia-induced performance deficit without stimulating spontaneous activity. Vinpocetine antagonized nearly completely CAR blockade in the 1.25-5.0 mg/kg p.o. dose range, restoring the ratio of avoidance responses to normal level. Vincamine exerted activity at a dose of 20 mg/kg p.o. Dihydroergotoxine and nicergoline were effective at 10 mg/kg p.o., Piracetam and meclofenoxate showed moderate protective activity at 2,000 and 500 mg/kg p.o., respectively. For comparison, the effect of dopaminergic drugs of a different mechanism of action (amphetamine, apomorphine, bromocriptine, and methylphenidate) was also tested in a similar situation. Only amphetamine (at 1.0 mg/kg i.p.) and low doses of apomorphine (0.1 and 1.0 mg/kg) had a favorable effect against hypoxia-induced CAR blockade; this effect was accompanied by an increase in locomotor activity. On the basis of these data, the relatively simple behavioral method applied by us, protection of acquired conditioned response against the disruptive effect of hypoxia in SH rats, seems suitable to detect nootropic activity.     

Adrenergic receptor blocking agents: Effects on central noradrenaline and dopamine receptors and on motor activity

Phenoxybenzamine, but not phentolamine or propranolol, blocked central noradrenaline receptors in flexor reflex experiments on the rat spinal cord using the selective noradrenaline receptor stimulating agent clonidine. None of these three drugs blocked dopamine receptors in experiments on turning of unilaterally striatectomized rats induced by the dopamine receptor stimulating agent apomorphine. The -methyltyrosine-induced disappearance of noradrenaline in the central nervous system of rats and mice was accelerated by phenoxybenzamine at doses related to the functional changes, whereas the other drugs were inefficient. The disappearance of dopamine was decelerated by phenoxybenzamine, but not by phentolamine or propranolol.The effects of the adrenergic receptor blocking agents on motor activity were studied in a model in which clonidine potentiated the activation induced by apomorphine in reserpine-treated mice. Phentolamine (20 mg/kg) and propranolol (10 mg/kg) reduced the stimulation seen both after apomorphine alone and in combination with clonidine, indicating nonspecific sedative effects. Phentolamine (10 mg/kg) blocked the peripheral effects of clonidine but did not markedly diminish the activation induced by the receptor stimulants. Phenoxybenzamine (20 mg/kg) blocked the clonidine-induced potentiation without interfering with the apomorphine-induced stimulation and can thus be used as a blocking agent of central and peripheral noradrenaline receptors in behavioural experiments.     

Presynaptic alpha-adenoceptors: the depression of self-stimulation by clonidine and its restoration by piperoxane but not by phentolamine or phenoxybenzamine.

Depression of self-stimulation by clonidine has been ascribed to continuous direct stimulation of alpha-adrenoceptors with consequent disruption of reinforcement signals thought to be conveyed by noradrenergic pathways. This suggestion was tested by administration of alpha-receptor blocking agents (piperoxane, phentolamine and phenoxybenzamine, PBZ) differing in their affinity for pre- and post-synaptic receptor sites. Piperoxane in low doses (0.55-5.0 mg/kg) previously reported to cause specific blockade of pre-synaptic receptors implicated in negative feedback circuits, caused a significant increase in self-stimulation rate and strongly antagonized the depression of self-stimulation by clonidine (0.15 mg/kg). A larger dose of piperoxane (45 mg/kg) and graded doses of phentolamine and PBZ, affecting both pre- and post-synaptic receptors, depressed self-stimulation, and did not antagonize clonidine-induced depression of self-stimulation. It is concluded that depression of self-stimulation by clonidine may depend on clonidine-induced inhibition of NA release exerted via presynaptic receptors, and that the effect of clonidine is not necessarily evidence that noncontingent adrenergic stimulation disrupts reinforcement.     

Learning deficits in 4 weeks old offspring of the nursing mothers treated with the neuroleptic drug penfluridol

Summary The acquisition of a conditioned avoidance response (CAR) was investigated in rats of nursing mothers given penfluridol 1.0 mg/kg on days 1, 3 and 5 after delivery. The male litter-mates were tested for CAR acquisition 4 weeks after birth. The animals whose mothers had received penfluridol were markedly inferior in the CAR acquisition than rats of mothers given glucose. Furthermore, rats of penfluridol-treated mothers were found to be heavier than those of glucose-treated mothers when weighed at 8 weeks of age. However, there was no difference between the groups at 4 weeks of age. The possibility that penfluridol interacts with the development of catecholamine mechanisms in the brain at a sensitive developmental period, and thereby influences later performance, is discussed.     

Reversal of the 6-hydroxydopamine-induced suppression of a CAR by drugs facilitating central catecholaminergic mechanisms.

The intraventricular injection of 6-hydroxydopamine (200 mug) blocked a previously learned conditioned avoidance response (CAR) in rats. The administration of NE, DOPA, amphetamine, phenelzine, desipramine and clonidine induced the reappearance of the CAR. These results are in agreement with current hypotheses on the mechanism of action of 6-hydroxydopamine on the central nervous system.     

Antipsychotic activity of substituted gamma-carbolines

Several novel substituted gamma-carbolines were synthesized and examined in a series of in vitro and in vivo pharmacological tests to determine potential antipsychotic activity. Most compounds were orally active in blocking the conditioned avoidance response (CAR) in rats but did not antagonize apomorphine-induced stereotyped behavior. Compound 17 (Wy-47,384), a gamma-carboline with a 3-(3-pyridinyl)propyl side chain, was selected for development as an atypical antipsychotic agent because of its potent and selective profile in preclinical psychopharmacological tests. It blocked CAR in rats with an AB50 of 14 mg/kg po, showed weak affinity for the D2 receptor site (Ki = 104 nM), and showed differential potency in antagonizing apomorphine-induced stereotyped behavior (ED50 = 11 mg/kg ip) and climbing behavior (ED50 = 4 mg/kg ip). Such activities are suggestive of antipsychotic efficacy combined with a low potential for extrapyramidal side effect (EPS) liability.     

Modulatory effects of ascorbate, alone or with haloperidol, on a lever-release conditioned avoidance response task.

Pretreatment with ascorbate, a modulator of dopamine transmission in the striatum, enhances the ability of haloperidol, a dopamine antagonist, to induce catalepsy and block the motor-activating effects of amphetamine. The present study extended this line of work to a lever-release version of the conditioned avoidance response (CAR) task, which is highly sensitive to changes in striatal dopamine. Adult male rats were trained to avoid footshock by releasing a lever within 500 ms of tone onset. Ascorbate (100 and 1000 mg/kg, IP) or vehicle was tested either alone or in conjunction with haloperidol (0.01 and 0.05 mg/kg, SC). Compared to vehicle pretreatment, 1000 mg/kg ascorbate alone or in combination with haloperidol impaired CAR performance by increasing avoidance latency. Latency to escape footshock was not impaired, ruling out a generalized motor deficit. In contrast, 100 mg/kg ascorbate alone or in combination with haloperidol had no consistent effects on CAR performance, even at a haloperidol dose (0.005 mg/kg, SC) known to potentiate dopamine transmission by preferentially blocking autoreceptors. Collectively, these results support an antidopaminergic action of ascorbate on striatal function, but suggest that this effect requires relatively high systemic doses.     

Effects of single and repeated exposures to abate on rat behavior and cholinesterase activity.

Rats were injected i.p. with the organophosphate insecticide ABATE and tested over the next 16 days. Animals given 1000 mg/kg showed impaired performance of a previously conditioned avoidance response 6 days after injection but not 2, 8, 10, or 16 days after injection. No behavioral changes were observed in animals given 316 or 562 mg/kg. A subsequent experiment showed that the avoidance impairment in animals given 1000 mg/kg was accompanied by significant erythrocyte, plasma, and brain cholinesterase activity inhibition and decreased spontaneous motor activity. If administration of the same ABATE dose was distributed over 6 days (167 mg/kg/day), cholinesterase and motor activity depression was still evident but conditioned avoidance performance was unimpaired. The results were interpreted as differential behavioral adaption to repeated injections of ABATE.     

The neurotensin-1 receptor agonist PD149163 inhibits conditioned avoidance responding without producing catalepsy in rats

Agonists for neurotensin (NT)-1 receptors have produced antipsychotic-like effects in many animals, including reversal of prepulse inhibition deficits and psychostimulant-induced increases in spontaneous activity. The present study sought to provide a basic assessment of the putative antipsychotic effects of PD149163 in rats using a two way conditioned avoidance response task, which is highly validated for screening antipsychotic drugs, and an inclined grid assessment, which is used to assess extrapyramidal side effect liability. PD149163 (0.0625-8.0 mg/kg) significantly suppressed conditioned avoidance responding (CAR) following administration of a 1.0 or 8.0 mg/kg dose. PD149163 failed to significantly increase catalepsy scores. The typical antipsychotic drug haloperidol (0.01-1.0 mg/kg) significantly suppressed CAR at a 0.1, 0.3, and 1.0 mg/kg dose, and a significant increase in catalepsy scores was found at the 1.0 mg/kg dose. The atypical antipsychotic drug clozapine (2.5-10.0 mg/kg) also produced a significant inhibition of CAR, which occurred following administration of a 10.0 mg/kg dose. Clozapine failed to significantly increase catalepsy scores. Finally, d-amphetamine (1.0 mg/kg), serving as a negative control, failed to suppress CAR or increase catalepsy scores. These data further suggest that PD149163 may have atypical antipsychotic-like properties.     

Reversal of clonidine-induced hypotension by beta-adrenoceptor blocking drugs

The interaction between clonidine, propranolol and sotalol was investigated using conscious normotensive and spontaneously hypertensive rabbits as well as conscious normotensive and spontaneously hypertensive rats. Clonidine (20 micrograms/kg) administration into the marginal ear vein of rabbits produced persistent hypotension and bradycardia. In propranolol (0.5 mg/kg, i.v.)-pretreated animals, clonidine-induced hypotension was prevented. In rats, daily oral or subcutaneous clonidine as well as propranolol produced hypotension and bradycardia. Significant antagonism of the observed hypotensive effects resulted when clonidine was given to propranolol-pretreated animals or when propranolol was added to the treatment regimen of animals being maintained on clonidine. No antagonism between sotalol and clonidine was demonstrable. In view of the known central site of action of clonidine, and the failure of sotalol to antagonize clonidine-induced hypotension it would appear that the central nervous system is a possible site of the observed drug interaction.     

Differential effects of d- and l-propranolol on dopamine turnover stimulated by oxotremorine in striatal and mesolimbic areas of rat brain

The effects of l-propranolol, d-propranolol and clonidine on homovanillic acid (HVA) concentrations in the corpus striatum and nucleus accumbens of rats were studied under normal conditions and after treatment with oxotremorine or haloperidol. Thile propranolol and clonidine given alone had no significant effect on HVA levels in either area, l-propranolol (1--10 mg/kg) and clonidine (0.1 mg/kg), both significantly inhibited the elevation of striatal HVA, found 60 min after oxotremorine administration. Both l- and d-propranolol (2.5 mg/kg), reduced the effect of oxotremorine in the nucleus accumbens. Neither l-propranolol no clonidine affected the rise in HVA in either brain area seen after haloperidol. Our results suggest that propranolol may reduce cholinergic activation of dopaminergic pathways by two different mechanisms. One is stereospecific for the l-isomer and operates in the striatum and another, which is shared by both isomers, occurs in the nucleus accumbens.     

Antidepressant effect of carbamazepine--the role of dopaminergic and noradrenergic agents.

Fourteen days of treatment with 20 mg/kg/day carbamazepine (CBZ) reduced the immobility time in the behavioral "despair" test in rats. Two blockers of dopaminergic receptors: haloperidol (0.5 mg/kg) and sulpiride (100 mg/kg) antagonized the reduction of immobility caused by CBZ. The anti-immobility effect of CBZ was also counteracted by clonidine (0.1 mg/kg) an agonist of presynaptic alpha 2 adrenoreceptors and propranolol (5 mg/kg) beta-adrenolytic drug.     

Domoic acid: Neurobehavioral and neurohistological effects of low-dose exposure in adult rats

Adult rats treated IP with domoic acid at 0, 0.22, 0.65, or 1.32 mg/kg were tested for passive avoidance (PA), auditory startle (AS), or conditioned avoidance (CAR) behaviors. Clinical signs were observed only at the 1.32 mg/kg dose level. Within 24 h of dosing, rats surviving a dose of 1.32 mg/kg exhibited transient decreased body weight and exaggerated AS responding. Startle latency and habituation, PA, and CAR were not affected. Examination of brains from six rats per group revealed a subset (2/6) of animals receiving 1.32 mg/kg domoic acid with degenerating neurons in the hippocampal CA1/CA3 subregions and gliosis. The decreased body weight and increased startle suggest a hyperreactivity syndrome possibly related to neuronal degeneration in the hippocampus. In a separate experiment, domoic acid at an IP dose of 0.93 mg/kg was found to produce hypomotility in addition to a decrease in body weight. Both effects were reduced by pretreatment with scopolamine (2 mg/kg), but not with caffeine (30 mg/kg), indicating a possible cholinergic involvement in domoate's toxicity.     

Effects of (-)3-PPP on acquisition and retention of a conditioned avoidance response in the rat

The administration of (-)3-(3-hydroxyphenyl)-N-n-propylpiperidine (3-PPP) was found to partially, but significantly, suppress the acquisition (4–8 mg/kg IP) and performance (8–16 mg/kg IP) of a conditioned avoidance response (CAR) in male Sprague-Dawley rats. All statistically significant effects were observed within 2 h of injection. Furthermore, using a situation in which the CAR was dependent on a visual successive discrimination, it was shown that discriminative performance was unaffected, and that (-)3-PPP (12.5–25 mg/kg IP) but not (+)3-PPP, suppressed the CAR. When (-)3-PPP (6.25 mg/kg IP) was combined with haloperidol (0.1–0.4 mg/kg IP), additive effects on the CAR performance were observed. Considering these effects, and the doses of (-)3-PPP required to suppress the CAR performance, it is concluded that the effects obtained in the present experiments are primarily due to a blockade of postsynaptic DA receptors.     

Enhanced acute antihypertensive effect of propranolol in the absence of circulating epinephrine in the rat

beta-Adrenoceptor blocking agents might reduce blood pressure, in part, by blocking presynaptic beta-adrenoceptors. Absence of circulating epinephrine should then reduce the antihypertensive effect of propranolol. Biadrenalectomized Wistar-Kyoto rats were made hypertensive with methylprednisolone (20 mg/kg s.c. weekly), given for 2 weeks, and supplemented with deoxycorticosterone pivalate (10 mg/kg weekly). Sham-operated controls received the same treatment. Baseline weight, mean intraarterial blood pressure, and heart rate of the groups were the same. After propranolol (5 mg/kg s.c.) was administered to the unanesthetized rats, blood pressure fell within 90 min from 151 +/- 4 by 23 +/- 4 mm Hg (mean +/- SEM) in the adrenalectomized animals and from 153 +/- 4 by only 7 +/- 3 mm Hg in the sham-operated controls (p less than 0.001); heart rate fell by 91 +/- 13 beats/min in the adrenalectomized animals and by 40 +/- 11 beats/min in the controls (p less than 0.01). Propranolol's vehicle had no effect. At the end of the experiment, plasma epinephrine levels were less than 40 pg/ml for adrenalectomized rats, and 420 +/- 60 pg/ml for controls. Norepinephrine levels were approximately equal in the two groups. Since blood pressure fell despite virtual absence of circulating epinephrine, these results suggest that propranolol reduces blood pressure, at least in part, by mechanisms other than presynaptic beta-adrenoceptor blockade.     

Effect of post-trial injection of beta adrenergic blocking agents on a conditioned reflex in rats

The effect of beta adrenergic blocking agents (propranolol, pronethalol, INPEA and dichloroisoproterenol) on the acquisition of a conditioned reflex was studied in adult, male Wistar rats. The experiments were done in a shuttle box with light as a conditioned stimulus, and an electric shock to the legs through the grid as the unconditioned stimulus. A maximum of 23 sessions were carried out with 24 h intervals. The drugs, the peripheral effects of which are estimated to last less than 24 h, were injected i.p. post-trial.The learning process and the number of animals which succeeded in complying with the learning criterion until session 23 were improved by propranolol (2 mg/kg) and pronethalol (10 mg/kg). INPEA (10 mg/kg) only increased the percentage of animals which succeeded in complying the learning criterion until session 12. In the initial sessions dichloroisoproterenol (10 mg/kg) impaired the learning process.Fellow of the Consejo Nacional de Investigaciones Cientificas y Técnicas, Argentina.     

Lesion of serotonergic neurons antagonizes clonidine-induced suppression of avoidance behavior and locomotor activity in rats

The effects of clonidine on avoidance acquisition and locomotor activity were studied in male Wistar rats with 5,6-dihydroxytryptamine (5,6-DHT) lesions of the median raphe nucleus. Lesioned animals showed marked depletion in forebrain serotonin and 5-hydroxyindole acetic acid concentrations. Clonidine (0.2 mg/kg IP in a single daily dose for 6 consecutive days) inhibited avoidance acquisition and reduced locomotor activity in unlesioned rats. In 5,6-DHT rats clonidine failed to produce depressive effects. The resistance of raphe-lesioned rats to clonidine is discussed on the basis of possible interaction between noradrenergic and serotonergic brain systems.     

Amphetamine,cocaine, and dizocilpine enhance performance on a lever-release,conditioned avoidance response task in rats

A lever-release version of the conditioned avoidance response (CAR) task was used to assess the behavioral effects of several psychomotor stimulants in rats. The indirect dopamine agonists,d-amphetamine (0.1 and 0.25 mg/kg) and cocaine (7.5 and 15 mg/kg), enhanced performance on this task. Both drugs incre ased percent avoidance responses and decreased avoidance latency. A higher dose of amphetamine (0.5 mg/kg) also decreased avoidance latency but failed to improve percent avoidance. Similar effects were seen at low (0.01 and 0.025 mg/kg) and high (0.05 mg/kg) doses of dizocilpine (MK-801), a stimulant that acts as a noncompetitive antagonist ofN-methyl-d-aspartate (NMDA) glutamate receptors. When combined with haloperidol (0.1 mg/kg), a dopamine antagonist, amphetamine (0.25 mg/kg) and dizocilpine (0.025 mg/kg) had differential effects on the lever-release CAR task. Thus, amphetamine-haloperidol was significantly better than haloperidol alone on percent avoidance but not on avoidance latency, whereas dizocilpine-haloperidol had the opposite effect: significantly better than haloperidol alone on avoidance latency but not on percent avoidance. Taken together, these results provide further support for dopaminergic mechanisms in CAR performance but suggest an opposing glutamatergic influence.