The potentiating effects of 4-aminopyridine on adrenergic transmission in the rabbit vas deferens.

The effects of 4-aminipyridine on adrenergic transmission in rabbit vas deferens were investigated. 10(-5)-10(-3) M 4-aminopyridine markedly potentiated the sustained, secondary contractile response to transmural stimulation and significantly increased the amount of (+/-)-|3H]-metaraminol released at 5 Hz, but had little effect on responses to exogenous (-)-noradrenaline. 10(-5)-10(-4) M 4-aminopyridine did not significantly alter the accumulation or efflux of (-)-[3H]-noradrenaline in rabbit heart. It is suggested that 4-aminopyridine potentiated responses of rabbit vas deferens to transmural stimulation mainly by increasing transmitter release, possibly as a result of prolongation of the action potential. Responses to transmural stimulation were also potentiated by by 2- and 3-aminopyridine and by 3,4-diaminopyridine but not by pyridine, aniline, 4-dimethylaminopyridine, 4-aminoethylpyridine or 2-aminopyrimidine.     

Clonidine inhibits electrodermal potentials induced by preganglionic stimulation

The electrodermal potential (EDP) recorded with surface electrodes between the palm and the shaven back of the right forepaw of anaesthetized and vagotomized cats was taken as a measure of the activity of cholinergic-sympathetic sudomotor nerves. EDPs were induced by preganglionic electrical stimulation of the stellate ganglion for 2 s with trains of DC pulses (2 ms duration, 0.5-128 Hz) at regular intervals of 60 s. The EDPs amounted to 12 mV and increased little with stimulation rate (14 mV). The i.v. injection of 30 micrograms/kg of the alpha 2-adrenergic agonist clonidine did not change the EDPs significantly. A consistent result was obtained in cats pretreated i.p. with 5 mg/kg reserpine 18 h beforehand for depletion of catecholamines. Three hours after the i.v. injection of 3 mg/kg guanethidine, clonidine (30 micrograms/kg i.v.) induced significant reduction of EDPs in the lower range of the stimulation rate but did not affect those at 16 and 32 Hz. Partial blockade of ganglionic nicotinic receptors by i.v. infusion of 0.08-0.3 mg/kg per min hexamethonium diminished EDPs (1 Hz) by 30-50%. Under these conditions the i.v. injection (30 micrograms/kg) or topical application of 1 microgram clonidine to the right stellate ganglion inhibited EDPs at all rates of stimulation. The inhibitory effects of clonidine could be antagonized by 200 micrograms/kg yohimbine i.v. Partial ganglionic blockade by i.v. infusion of the depolarizing blocker suxamethonium (0.2-0.4 mg/kg per min) decreased EDPs. However, the topical application of 1 microgram clonidine to the stellate ganglion during infusion of suxamethonium caused no further decrease.(ABSTRACT TRUNCATED AT 250 WORDS)     

Involvement of spinal alpha adrenoceptors in mediation of the hypotensive action of guanabenz in rats

Summary The effects of intrathecal, intracisternal and intravenous injections of yohimbine (10 g) on hypotensive and bradycardic responses to the alpha agonist guanabenz (10 g/kg, i.v.) were studied in anesthetized rats. The depressor response to guanabenz was inhibited by intrathecal pretreatment with yohimbine, while this pretreatment did not affect the bradycardic response to guanabenz. Intracisternal pretreatment with yohimbine inhibited both cardiovascular responses to guanabenz, whereas intravenous pretreatment with yohimbine affected neither. Guanabenz (1 g) decreased blood pressure when injected intrathecally but did not affect it when injected intravenously. These results suggest that in rats spinal alpha adrenoceptors are involved in mediation of the hypotensive action of the alpha agonist guanabenz injected systemically. Send offprint requests to T. Kubo     

Studies on the centrally mediated hypotensive activity of guanabenz.

Prior studies demonstrated that guanabenz reduces systemic blood pressure by inhibiting central sympathetic outflow as well as by adrenergic neuron blockade. Potential mechanisms responsible for the reduction of efferent sympathetic activity were examined in the present series. Guanabenz failed to modify carotid sinus nerve activity in a perfused sinus preparation. It reduced sympathetic outflow, heart rate and blood pressure in debuffered cats indicating that its actions are not mediated primarily by baroreceptor mechanisms. alpha-Adrenergic blockade greatly attenuated the response suggesting that the central sympathoinhibitory effect of guanabenz results from alpha-adrenergic receptor activation. Only a high dose of the compound attenuated the increase in sympathetic nerve activity produced by stimulation of the posterior hypothalamus. These experiments lead to the overall conclusion that guanabenz acts primarily at sites which regulate the basal level of sympathetic outflow.     

A quantitative assessment of CNS sympatho-inhibition produced by psychotropic drugs

As one index of sympathetic reactivity, electrodermal responses (EDR) were evoked from central (hypothalamic) and peripheral (ulnar nerve) sites in pentobarbital-anesthetized cats. When compared with intravenous chlorpromazine (ED₅₀ ～ 1.0 mg/kg), only thioridazine, trifluoperazine and pimozide were less potent than chlorpromazine in reducing the amplitude of these centrally-evoked sympathetic-cholinergic responses. Perphenazine and methotrimeprazine (a non-neuroleptic phenothiazine) were about twice as potent as chlorpromazine. Haloperidol and triflupromazine were about 5 times as potent and chlorprothixine was more than 10 times as potent. None of these agents reduced the peripherally-evoked electrodermal response, indicating a CNS mode of action. Diazepam was without effect at either site. In addition, pretreatment with yohimbine (0.5 mg/kg, i.v.) did not significantly alter the ED₅₀ for any of the above drugs. These results demonstrate that all of the phenothiazines and non-phenothiazine neuroleptics tested produced a dose-dependent central sympatho-inhibition and that diazepam does not. The results also suggest that there is no significant correlation between central symphatho-inhibition and the antipsychotic potency of these compounds and that their depression of central sympathetic outflow is independent of α-adrenergic mechanisms in the CNS.     

Comparison of guanabenz and clonidine in hypertensive patients

Summary

The antihypertensive and clinical effects of two centrally acting drugs, guanabenz and clonidine, were compared in a double-blind trial in 29 patients with established hypertension. After a 1-week baseline period and 2 weeks on placebo, patients received treatment with either guanabenz (mean dose 24?mg daily) or clonidine (mean dose 0.45?mg daily) alone for 8 weeks. Both drugs produced equivalent and highly significant (p < 0.001) reductions in systolic and diastolic blood pressures in the standing and supine positions. They also reduced significantly the standing and supine pulse rates. Normal orthostatic responses were maintained with both regimens. AN but 1 patient in each group reported side-effects during active treatment, the most frequent being dry mouth and sedation with each drug. No laboratory or ECG abnormalities related to treatment were observed.     

Effect of clonidine and chlorpromazine on centrally evoked electrodermal responses and their interaction with yohimbine

Both clonidine and chlorpromazine reduced the amplitude of electrodermal responses (EDR) evoked by stimulation of the hypothalamus at a constant submaximal frequency (10–16 Hz). The ED₅₀ for clonidine was approximately 5 μg/kg and that for chlorpromazine was about 1 mg/kg. Yohimbine pretreatment (0.5 mg/kg, i.v.) antagonized the effects of clonidine but did not alter the effectiveness of clorpromazine in inhibiting these responses. Yohimbine alone was without effect on these sympathetic-cholinergic responses. These results suggest that clonidine and chlorpromazine depress central sympathetic reactivity by different mechanisms.     

A physiological and pharmacological analysis of the electrodermal response in the rat

Electrodermal responses (EDR) of the sympathetic-cholinergic sudomotor system were elicited in the footpads of the hindpaws of anesthetized rats. The most reactive CNS loci were caudal to the region of the posterior hypothalamus. Peripherally evoked responses were elicited by electrical stimulation of the decentralized tibial nerve. The amplitude of these evoked EDR was stable over time and both the centrally and peripherally elicited responses were frequency-dependent. Atropine (200 μg/kg) depressed the EDR elicited by both peripheral and central stimulation whereas hexamethonium (20 mg/kg) only inhibited the central EDR. Clonidine had no effect on the EDR evoked at the periphery but produced a significant dose-dependent depressant effect on the centrally evoked EDR; this effect was partially antagonized by yohimbine (0.75 mg/kg). It is suggested that the rat is a suitable species for the use of the sudomotor system in the investigation of adrenergic agents which are thought to have a central sympatho-inhibitory action.     

Studies on the site of action of clonidine utilizing a sympathetic-cholinergic system

Clonidine (30 μg/kg, i.v.) reduced centrally evoked electrodermal responses (EDR) following stimulation of reactive loci in the hypothalamus and medulla. The responses were most depressed following low frequency stimulation. Similar results were observed on the EDR evoked by stimulation of the cervical cord in the spinal act. Little effect was seen following peripheral nerve stimulation. These results demonstrate that clonidine depresses the reactivity of this sympathetic-cholinergic system at all central levels including the cervical cord.     

Topical clonidine produces mydriasis by a central nervous system action.

One drop of clonidine solution (0.125, 0.25 or 0.5%) was administered topically to one eye in cats anesthetized with pentobarbital in which the vagosympathetic nerves had been sectioned. Clonidine caused a simultaneous dose-related mydriasis in both eyes along with a decrease in heart rate. The peak effects were observed in about 20--30 min. Topical administration of clonidine (0.5%) produced no effect on the parasympathectomized, eserinized iris but did dilate the opposite pupil. Epinephrine (0.1--30 microgram, i.a.) produced equal pupillary dilation in both eyes. In addition, topical clonidine caused a dramatic decrease in postganglionic ciliary nerve activity. All of the effects of clonidine were antagonized by yohimbine (0.5 mg/kg, i.v.). These results demonstrate that topical administration of clonidine causes my driasis in the cat and that this effect is mediated totally by means of CNS inhibition of parasympathetic tone to the iris.     

Analysis of pupillary dilation produced by analogs of clonidine

Pupillary responses to intravenous administration of clonidine and five congeneric derivatives of clonidine were observed in anesthetized cats. All of the agents tested produced a dose-dependent mydriasis of long duration. The order of potency for these compound was clonidine (St-155) > St-375 > St-606 > St-600 > St-608 > St-91. Pretreatment with yohimbine hydrochloride (0.5 mg/kg i.v.) shifted the dose-response curve for all of the compounds tested to the right. A CNS inhibition of parasympathetic nerve activity was demonstrated for clonidine and St-375 by means of direct recordings from the postganglionic ciliary nerves. An effect of high doses of St-91 on nerve activity was also observed. These results suggest that all the analogs of clonidine tested (with the possible exception of St-91) act like clonidine to produce mydriasis by a central α-adrenergic inhibition of parasympathetic outflow to the eye.     

Cardiovascular effects of alpha-adrenergic drugs: Differences between clonidine and guanabenz

Summary Guanabenz induced a pressor effect in pithed rats through postsynaptic ₂-adrenoceptors whereas clonidine activated both vascular ₁ and ₂-adrenoceptors. Previous treatment with prazosin, and ₁-antagonist, or depletion of the noradrenergic stores by reserpine produced supersensitivity to the pressor response to clondine only, probably through postsynaptic ₁-adrenoceptors.The hypotension and bradycardia developed in normotensive rats after intravenous guanabenz administration were abolished by prazosin, whereas the central effects of clonidine were antagonized by both prazosin and yohimbine.Selective destruction of central noradrenergic neurons by [N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine] (DSP 4) or reserpine plus blockade of catecholamine synthesis by -methyl-p-tyrosine abolished the hypotension and bradycardia produced by guanabenz but merely reduced the bradycardia from clonidine.The present results suggest that, in rats, guanabenz is a selective stimulant of central -autoadrenoceptors antagonized by prazosin whereas at a vascular level guanabenz preferentially activates -adrenoceptors antagonized by yohimbine. The differences observed between the mechanisms by which guanabenz and clonidine produce their central cardiovascular responses might be attributed to their acting on different nuclei.     

Central sympathetic reactivity inhibited by indoramin

The action of indoramin on central autonomic activity was investigated using the sympathetic-cholinergic electrodermal response system. Indoramin (0.33–10 mg/kg, i.v.) significantly reduced the amplitude of electrodermal responses (EDR) evoked by stimulation of the hypothalamus in a dose-related manner. The CNS effects of indoramin on the EDR were partially antagonized by yohombine pretreatment. These results demonstrate that indoramin reduces central sympathetic reactivity and suggest that an α-adrenergic inhibitory mechanism may be involved.     

Clonidine inhibits nicotinic effects in ganglia of the cholinergic-sympathetic system

The electrodermal potential (EDP) recorded with surface electrodes between the palm and the shaven back of the forepaws of anaesthetized cats was taken as a measure of sudomotor activity in response to submaximal activation of cholinergic-sympathetic efferents with the nicotinic ganglionic stimulant DMPP. The intravenous injection of 30 micrograms/kg clonidine inhibited EDPs in despinalized cats and in vagotomized cats with chronic denervation of the stellate ganglion. In vagotomized cats clonidine (30 micrograms/kg i.v.) shifted the dose-response curve of DMPP (4-256 micrograms/kg i.v.) to the right. An inhibitory effect was also observed in despinalized cats upon topical application of 0.1 and 1 microgram clonidine to the stellate ganglion. In all preparations the inhibitory effect of clonidine could be antagonized by intravenous injection of 200 micrograms/kg yohimbine. As all substances used were also active when injected during arrest of blood flow to the foreleg by means of a tourniquet, actions at the level of the sweat gland could be excluded. The results lead to the conclusion that the activation of postsynaptic somadendritic alpha 2-adrenoceptors in sudomotor ganglia of the cholinergic-sympathetic nervous system inhibits the effects of nicotinic ganglionic stimulation. It is assumed that this action is due to the known ganglionic hyperpolarization induced by alpha 2-adrenoceptor stimulants.     

Effects of clonidine and chlorpromazine on a sympathetic-cholinergic reflex

Intravenous administration of clonidine and chlorpromazine resulted in a dose-dependent inhibition of the amplitude of reflexly evoked electrodermal responses in intact and spinal cats. Yohimbine pretreatment (0.5 mg/kg, i.v.) antagonized the effects of clonidine but not chlorpromazine in both preparations. These findings confirm and expand previous observations that both clonidine and chlorpromazine inhibit the amplitude of centrally evoked responses in this sympathetic-cholinergic system. In addition, both drugs appeared to have a spinal site of action. The antagonism of the effects of clonidine by yohimbine suggests that the mechanism of the action of clonidine may be a result of activation of central inhibitory alpha-adrenergic receptors. The failure of yohimbine to antagonize the effects of chlorpromazine suggests that clonidine and chlorpromazine may depress these sympathetic reflexes by different mechanisms.     

Differential effects of d,l-amphetamine on licking maintained by electrical hypothalamic stimulation and/or water in rats

The role of the reinforcer as a determinant of the behavioural effects of amphetamine was assessed with a procedure under which response requirements were equated. Eight rats were trained to lick on fixed-ratio schedules of rewarding lateral hypothalamic stimulation and water delivery, respectively. The effects of d,l-amphetamine (0.1–2.0 mg/kg) were investigated in the lateral hypothalamic stimulation condition and compared with the effects of the drug at 1.0 mg/kg in the water condition. Licking maintained by hypothalamic stimulation was increased after amphetamine administration to approximately 200% of control performance at 1.0 mg/kg. At this dose amphetamine administration resulted in a decrease in water maintained licking. A computer analysis of the temporal patterning of licking in both water and stimulation conditions was carried out. This analysis revealed that amphetamine may attenuate the disruptive effects of hypothalamic stimulation on the lick-interrupt cycle. The differential effects of amphetamine on licking maintained by the events may reflect either unequal participation of catecholaminergic circuits in the two types of reward, or anti-inhibitory motor effects of amphetamine.     

Action of enpiprazole on emotional behavior induced by hypothalamic stimulation in rats and cats

Action of enpiprazole on emotional behavior elicited by hypothalamic stimulation in rats and cats was investigated and comparisons were made with effects of diazepam. Two behavioral patterns were elicited by stimulation of the postero-medial part of the hypothalamus in rats: a food-carrying response beginning with exploratory movement and an analogue of fear. Enpiprazole frequently changed the food-carrying response into food-taking response and occasionally analogues of fear into food-carrying and/or food-taking responses. Thresholds for these behaviors were also elevated. Diazepam showed the same effects on the thresholds as enpiprazole, having but little effect on the behavioral patterns.In cats, enpiprazole elevated the thresholds for affective-defensive responses induced by hypothalamic stimulation in 6 of 8 cases, but lowered them in 2 cases. This suggests that enpiprazole has a biphasic effect in the central nervous system. By contrast, diazepam consistently elevated thresholds. Comparing the action of enpiprazole with that of diazepam it can be presumed that the former is a different type of anxiolytic drug than the latter.     

One-way generalization of clonidine to the discriminative stimulus produced by cocaine

Rats were trained to discriminate the stimulus properties of either cocaine or clonidine using a food reinforced two-lever choice paradigm. After training, cocaine was generalized to the cocaine lever in a dose-dependent manner, and clonidine was generalized to the clonidine lever in a dose-dependent manner. Yohimbine, an alpha-2 antagonist, blocked the clonidine stimulus but not the cocaine stimulus. Cocaine was not generalized to the clonidine stimulus; however, clonidine was generalized to the cocaine stimulus, and this generalization was blocked by yohimbine. The one-way generalization of clonidine to cocaine suggests that clonidine has at least two discrete stimulus components: a major component that is not cocaine-like, and a minor component that can be detected by cocaine-trained subjects. In addition, the yohimbine blockade data suggest that both components of the clonidine stimulus are mediated via alpha-2 receptors.     

The effects of alaproclate on the pupillary responses to tyramine,phenylephrine and pilocarpine in depressed patients

Nine depressed patients were treated with alaproclate, a selective 5-HT uptake inhibitor, for 3 weeks in a dose of 400 mg daily. The pupillary responses to tyramine, phenylephrine, and pilocarpine eye drops were measured on consecutive days before, after 1 week and after 3 weeks of treatment. The tyramine-induced mydriasis was unaffected by alaproclate, suggesting that it does not significantly inhibit the reuptake of noradrenaline. The pilocarpine-induced miosis and the phenylephrine-induced mydriasis were both enhanced after 1 week but not after 3 weeks of treatment. This suggests that alaproclate acutely increases the responsiveness of postsynaptic muscarinic and ₁ adrenoceptors.     

Effects of yohimbine on CNS structures: Neurophysiological and behavioral correlations

Yohimbine (by intravenous and/or intraperitoneal routes at doses between 0.5 to 3 mg/kg) produces behavioral changes (in freely moving cats and dogs) characterized by: a) an increase in alertness, extreme agitation, and nervousness; b) an increase in motor reaction to a variety of sensory stimuli; and c) an appearance of muscular shaking and tremors. These behavioral changes are associated with electrographic alterations characterized by: a) an increase in the amplitude of the local evoked potentials (LEP) recorded from the mesencephalic reticular formation, posterior hypothalamus and intralaminar thalamic system; b) a decrease in the amplitude of the LEP recorded from preoptic area, lateral hypothalamus and septal region; c) a slight diminution in the LEP amplitudes of the amygdala and hippocampus; d) an initial increase (15 to 25 min) followed by a long period of strong diminution in the amplitude of the cortical LEP; e) an increase in the percent time of fast cortical activity and in the amplitude of all frequencies; f) an appearance of subcortical and cortical epileptiform discharges in the normal animals during administration of the compound at doses of 2 to 3 mg/kg; and g) a marked activation of epileptiform discharges in the subcortical or cortical epileptic animals in association with occasional clinical seizures. These results were discussed from the point of view of neuronal excitability changes in CNS structures.This project was supported in part by the Louisiana State Department of Hospitals, and in part by a USPHS Grant, Number 2-R10-MH-03701-13, Psychopharmacology Research Branch, NIMH, U.S.A.