Central Alpha 2-adrenoceptors Research Articles

Chronic subordination stress in male tree shrews: Replacement of testosterone affects behavior and central alpha 2-adrenoceptors

Subordination stress induced by social defeat in male animals is known to inhibit gonadal functions and it has been discussed whether the resulting deficit in testosterone might play a role in subordination behavior. One of the major transmitter systems involved in regulation of behavior is the noradrenergic system. To analyze whether a testosterone replacement can alter subordination behavior and whether this might be related to changes in the brain noradrenergic system, we quantified alpha 2-adrenoceptors (alpha 2-ARs) in the central nervous system of male tree shrews. Animals were submitted to chronic subordination stress and received testosterone at the same time. Behavior was monitored during all phases of the experiment: the control period of 10 days, the period of social stress lasting 10 days when subordinates were confronted daily with a dominant male, and, subsequently, the stress and treatment period of 18 days when in parallel to the stress, animals received either injections of testosterone or vehicle. Brain alpha 2-ARs were quantified by in vitro receptor autoradiography using the antagonist ligand 3H-RX821002. Locomotor activity decreased significantly during the stress period and was not re-normalized by testosterone. In contrast, testosterone re-normalized scent marking behavior and autogrooming, parameters that had both been reduced due to the subordination stress. Vehicle injections improved none of these behaviors. In 8 of 10 brain regions that were analyzed, numbers of alpha 2-adrenergic binding sites were increased in stressed animals that received vehicle injections, but a difference between testosterone and vehicle injected animals was only observed in five regions. These brain regions are all known to be involved in emotional behavior (anterior hypothalamus, medial nucleus of the amygdala, cingulate cortex) or autonomic regulation, respectively (solitary tract nucleus, dorsal motor nucleus of vagus). Therefore, our data show that testosterone influences behavior of male subordinates and modulates alpha 2-AR expression in their brains. Androgen-mediated alterations in receptors occur in brain regions that are known to be involved in emotionality, e.g., in the anterior hypothalamus which regulates aggressive behavior. One can therefore conclude that alpha 2-ARs contribute to neuronal functions that are responsible for subordination of stress behavior, and that testosterone-induced receptor changes are related to the partial restoration of normal behavior.

Respective contributions of alpha-adrenergic and non-adrenergic mechanisms in the hypotensive effect of imidazoline-like drugs.

The hypotensive effect of imidazoline-like drugs, such as clonidine, was first attributed to the exclusive stimulation of central alpha2-adrenoceptors (alpha2ARs). However, a body of evidence suggests that non-adrenergic mechanisms may also account for this hypotension. This work aims (i) to check whether imidazoline-like drugs with no alpha2-adrenergic agonist activity may alter blood pressure (BP) and (ii) to seek a possible interaction between such a drug and an alpha2ARs agonist alpha-methylnoradrenaline (alpha-MNA). We selected S23515 and S23757, two imidazoline-like drugs with negligible affinities and activities at alpha2ARs but with high affinities for non-adrenergic imidazoline binding sites (IBS). S23515 decreased BP dose-dependently (-27+/-5% maximal effect) when administered intracisternally (i.c.) to anaesthetized rabbits. The hypotension induced by S23515 (100 microg kg(-1) i.c.) was prevented by S23757 (1 mg kg(-1) i.c.) and efaroxan (10 microg kg(-1) i.c.), while these compounds, devoid of haemodynamic action by themselves, did not alter the hypotensive effect of alpha-MNA (3 and 30 microg kg(-1) i.c.). Moreover, the alpha2ARs antagonist rauwolscine (3 microg kg(-1) i.c.) did not prevent the effect of S23515. Finally, whilst 3 microg kg(-1) of S23515 or 0.5 microg kg(-1) of alpha-MNA had weak hypotensive effects, the sequential i.c. administration of these two drugs induced a marked hypotension (-23+/-2%). These results indicate that an imidazoline-like drug with no alpha2-adrenergic properties lowers BP and interacts synergistically with an alpha(ARs agonist.

Involvement of the opioid system in the central antisecretory action of alpha-2 adrenoceptor agonists in rat

The aim of the present study was to analyse the role of the central alpha-2 adrenoceptors in the regulation of gastric acid secretion in pylorus ligated rats. It was found that the intracerebroventricularly (icv.) injected presynaptic alpha-2 adrenoceptor agonist clonidine and the alpha-2A adrenoceptor subtype selective stimulant oxymetazoline exerted a dose dependent inhibition on gastric acid secretion. The antisecretory ED 50 values for clonidine and oxymetazoline were 20 and 7.5 nmol/rat icv., respectively. The antisecretory effect of these compounds was antagonised by the presynaptic adrenoceptor antagonist yohimbine (50 nmol/rat icv.) indicating that the action is mediated through central presynaptic alpha-2 adrenoceptors. Moreover, naloxone (50 nmol/rat icv.)—non-selective opioid antagonist—and naltrindole (0.5 nmol/rat icv.)—delta-opioid receptor selective antagonist—also decreased the antisecretory effect of clonidine and oxymetazoline suggesting that the endogenous opioid system is likely to be involved in the central antisecretory action of alpha-2 adrenoceptor stimulants.

Galanin/alpha2-adrenoceptor interactions in telencephalic and diencephalic regions of the rat.

The aim of this study was to evaluate whether galanin could affect central alpha2-adrenoceptors in telencephalic and diencephalic regions in the rat using quantitative receptor autoradiography with the alpha2 agonist radioligand [3H]p-aminoclonidine. Galanin 1 nM significantly and substantially increased the Kd value of the [3H]p-aminoclonidine binding sites in the medial hypothalamus and amygdala by 86% (p < 0.01) and 73% (p < 0.05) respectively. The Bmax value was only significantly increased with 3 nM galanin in the amygdala and the medial hypothalamus (both p < 0.05). The antagonist M35 counteracted the increase of the Kd values of the alpha2-adrenoceptor agonist binding sites produced by galanin 1 nM in the amygdala and the medial hypothalamus (both p < 0.001). These findings suggest the existence of an antagonistic galanin/alpha2 adrenoceptor interaction in the medial hypothalamus and amygdala that may be of relevance for alpha2-adrenoceptor-regulated neuroendocrine functions and food intake.

Enhancement of antipsychotic-like effects by combined treatment with the alpha1-adrenoceptor antagonist prazosin and the dopamine D2 receptor antagonist raclopride in rats.

Blockade of central alpha1-adrenoceptors has been implicated as a possible factor contributing to the atypical antipsychotic profile of clozapine. Thus, in the present study we examined the effects of concomitant alpha1-adrenoceptor and dopamine D2 receptor blockade on conditioned avoidance response performance, as an index of antipsychotic-like activity, and on the induction of catalepsy, as a test for extrapyramidal side effect liability, in rats. It was found that pretreatment with the alpha1-adrenoceptor antagonist prazosin (0.2mg kg(-1) s.c.) caused an enhancement of a suppression of conditioned avoidance response in the presence of the dopamine D2 receptor antagonist raclopride (0.05-0.20 mg kg(-1) s.c.). The effect was most prominent at a subthreshold dose of raclopride (0.05 mg kg(-1)). At these doses, prazosin or raclopride by themselves, or in combination, did not produce catalepsy. In addition, pretreatment with prazosin (0.2mgkg(-1) s.c.) did not alter the catalepsy produced by a higher dose of raclopride (1.0 mg kg(-1) s.c.). It is suggested that, in the presence of low dopamine D2 receptor occupancy, additional alpha1-adrenoceptor blockade might improve antipsychotic efficacy, and thereby improve the therapeutic window with regard to parkinsonism.

Actions of L-NAME and methylene blue on the hypotensive effects of clonidine and rilmenidine in the anesthetized rat.

The antihypertensive mechanism of alpha2-adrenoceptor agonists, such as clonidine and rilmenidine, is not completely elucidated, although it is probably due to reduction of sympathetic tone mediated by stimulation of central alpha2-adrenoceptors. Because activation of alpha2-adrenoceptors on endothelial cells induces release of endothelium-derived relaxing factor (EDRF), we determined whether nitric oxide (NO) release is involved in the antihypertensive action of clonidine and rilmenidine. In chloralose-anesthetised Wistar rats, systolic and diastolic arterial blood pressures were recorded on a polygraph. Intravenous injection of clonidine or rilmenidine (control group) caused a rapid increase of arterial blood pressure. followed by a long-lasting hypotensive effect. The hypotensive effects, estimated as the area enclosed by the decrease in diastolic pressure during the 20 min after clonidine and rilmenidine injections, were 574+/-60 and 410+/-59 mm Hg/min, respectively. The delta decrease in diastolic arterial blood pressure observed 20 min after intravenous injections of clonidine and rilmenidine was 48+/-5 and 34+/-3 mm Hg, respectively. Clonidine and rilmenidine injected 5-10 min after intravenous pretreatment with L-NAME (2 and 1 mg/kg) or methylene blue (10 mg/kg) induced hypotensive effects that were significantly smaller than that observed for the control group. These results suggest that the antihypertensive effects of clonidine and rilmenidine also may be modulated by the NO-cyclic guanosine monophosphate (cGMP) pathway at the level of the central nervous system and/or at the vascular peripheral circulation.

Intracerebroventricular injection of clonidine releases β-endorphin to induce mucosal protection in the rat

The possibility that the endogenous opioid system could be involved in the central nervous system (CNS)-mediated gastroprotective effect of clonidine was investigated. Intracerebroventricularly (i.c.v.) injected clonidine (470 pmol/rat) inhibited the gastric mucosal lesions induced by (orally administered) acidified ethanol in a significant manner in the rat. The gastroprotective effect of the centrally administered clonidine was antagonised by i.c.v. or intracisternally (i.c.) administered presynaptic alpha-2 adrenoceptor antagonist, yohimbine; the non-selective opioid receptor antagonist, naloxone; and the delta opioid receptor antagonist naltrindole. These results suggest that an interaction between central alpha-2 adrenoceptors and endogenous opioid systems is involved in mediating the mucosal protective effect. β-endorphin antiserum (i.c.) also antagonised the gastroprotection induced by intracerebroventricularly injected clonidine indicating that β-endorphin release is likely to be a key factor in the gastroprotective effect of clonidine. Furthermore, the i.c.v. or i.c. injection of beta-endorphin produced a potent gastroprotection in the picomolar range. The mucosal protective effect of clonidine was abolished after vagotomy indicating that the central effect may be conveyed to the periphery by vagal efferents. Since atropine (1 mg/kg i.v.) failed to modify, but hexamethonium (10 mg/kg i.v.) antagonised the gastroprotective effect of clonidine, it would appear that in the periphery nicotinic, but not muscarinic, cholinergic receptors are likely to be involved in the mucosal protective effect of clonidine. In conclusion, clonidine (i.c.v.) induces gastroprotective action by releasing an endogenous opioid substance — most likely β-endorphin — in the rat. The clonidine-induced central gastroprotection requires the integrity of vagal pathway; cholinergic nicotinic — but not muscarinic — receptors might mediate the effect in the periphery.

Brain alpha 1-adrenergic neurotransmission is necessary for behavioral activation to environmental change in mice

Terazosin, a water-soluble alpha 1 antagonist that can be administered in high doses intraventricularly was used to study the relationship between brain alpha 1 adrenoceptor neurotransmission and behavioral activation in the mouse. The antagonist was found to produce a dose-dependent, complete inhibition of motor activity and catalepsy which were reversed preferentially by coinfusion of an alpha 1 agonist (phenylephrine) compared to a D1 (SKF38393) or a D2 agonist, (quinpirole). Blockade of central beta-1 (betaxolol), alpha-2 (RX821002) or beta-2 (ICI118551) adrenoceptors had smaller or non-significant effects. Terazosin's selectivity for alpha 1 receptors versus dopaminergic receptors was verified under the present conditions by showing that the intraventricularly administered antagonist protected striatal and cerebral cortical alpha 1 receptors but not striatal or cortical D1 receptors from in vivo alkylation by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroxyquinoline. That its effect was due to blockade of brain rather than peripheral receptors was shown by the finding that intraperitoneal doses of terazosin three to 66 times greater than the maximal intraventricular dose produced less behavioral inhibition. Intraventricular terazosin also produced hypothermia and a reduced respiratory rate suggestive of a reduced sympathetic outflow. However, external heat did not affect the inactivity, and captopril, a hypotensive agent, did not mimic it. Terazosin did not impair performance on a horizontal wire test or the ability to make co-ordinated movements in a swim test suggesting that its activity-reducing effect was not due to sedation and may have a motivational or sensory gating component. It is concluded that central alpha 1-noradrenergic neurotransmission is required for behavioral activation to environmental change in the mouse and may operate on sensorimotor and motivational processes.

Central benzodiazepine involvement in clonidine cardiovascular actions

It is well known that the GABAergic and noradrenergic systems play an important role in blood pressure and heart rate regulation. Benzodiazepines and beta-carbolines, respectively, increase or decrease the probability of chloride-channel opening induced by GABA. The aim of this study was to determine, in conscious rats, the interaction existing between the central alpha2-adrenoceptor stimulation induced by clonidine and the facilitation or impairment of benzodiazepine receptor activity through the administration of either diazepam, a benzodiazepine receptor agonist, or methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM), an inverse benzodiazepine agonist. Clonidine (5-10 microg, intracerebroventricularly) reduced heart rate and increased mean blood pressure by activation of central alpha2-adrenoceptors. Diazepam (2 mg/kg, intravenously (i.v.)) induced an increase in heart rate, while DMCM (0.3 mg/kg, i.v.) elicited a bradycardic effect. The bradycardic effects induced by both clonidine and DMCM were antagonized by the prior administration of methylatropine (1.5 mg/kg, i.v.). DMCM (0.3 mg/kg, i.v.) prevented the clonidine effects on heart rate and mean blood pressure, while diazepam (2 mg/kg, i.v.) failed to modify these effects. Our results suggest that the bradycardic effects of clonidine are mediated by a vagal stimulation and are related to the activation of a GABAergic pathway.

Interactions of intravenous anesthetics with cerebral alpha-2-adrenoceptors

Up to date the exact mechanisms of action of anaesthetics on structures of the central nervous system have not been elucidated. Agents acting on central alpha 2-adrenoceptors have been demonstrated to possess potent sedative properties. Beside the classical agonists, general anaesthetics that are currently in clinical use e.g. etomidate have also been shown to act on alpha 2-adrenoceptors. Thus, the aim of this study was to study the effect of the intravenous anaesthetic agents propofol and ketamine on the binding of the specific alpha 2-adrenoceptor agonist paraiodoclonidine at cerebral alpha 2 adrenoceptors. After approval of the local animal care committee brain tissue of six Wistar rats was removed and frozen at -80 degrees C. The tissue was sectioned in 15-20 microns thick slices. After washing in TRIS-buffer (pH = 7.5) the slices were incubated with 2.5 nM 125I-paraiodoclonidine (PIC) for determination of specific binding. Unspecific binding was determined in presence of 200 microM unlabeled clonidine. Displacement experiments were performed in presence of propofol (11-530 microM) and ketamine (52-1100 microM) and evaluated autoradiographically. Average exposure time was three days. All films were then analysed by densitometry. Statistical significance calculated by Student's t-test was assumed at a level of p < 0.05. Only at the highest concentration of propofol and ketamine an effect on PIC binding was noticeable. Neither propofol nor ketamine was able to displace PIC completely from cerebral alpha 2-adrenoceptors. In contrast to other anaesthetics such as etomidate which was demonstrated to displace PIC completely from alpha 2-adrenoceptors, propofol and ketamine exerted only minor effects on the binding of PIC even at concentrations that exceeded clinical concentrations. These data suggest that an additional site of action of these anaesthetics at cerebral alpha 2-adrenoceptors beside the known actions on GABA receptors or NMDA receptors is unlikely.

Pilocarpine treatments differentially affect alpha2-adrenoceptors which modulate the firing rate of locus coeruleus neurones and the synthesis and release of noradrenaline in rat brain.

We have investigated the effect of treatments with the muscarinic acetylcholine receptor agonist, pilocarpine, on the sensitivity of central alpha2-adrenoceptors that regulate the firing activity of rat locus coeruleus, the tyrosine hydroxylase activity in the rat cortex, hippocampus and hypothalamus, and the K(+)-evoked release of [3H]noradrenaline from rat cortical and hippocampal synaptosomes. Short-term (4 days), but not acute, treatment with pilocarpine caused a small but statistically significant increase in the inhibitory effect of the alpha2-adrenoceptor agonist clonidine on the firing rate of locus coeruleus neurones, with a decrease in the ED50 of 290% (P<0.001). However, no change in the effect of clonidine on the locus coeruleus was observed after longer pilocarpine (11 days) treatment. In the rat cerebral cortex, but not in hippocampus or hypothalamus, chronic (19 days) treatment with pilocarpine caused a decrease in the inhibitory effect of clonidine on tyrosine hydroxylase activity (55%, P<0.05), but did not change the stimulatory effect of the alpha2-adrenoceptor antagonist idazoxan. Moreover, treatments (4, 11 and 19 days) with pilocarpine did not alter the inhibitory effect of clonidine [10(-8)-10(-5) M] on the K(+)-evoked release of [3H]noradrenaline from rat cortical and hippocampal synaptosomes. These results indicate that administration of pilocarpine slightly potentiates some but not all the functional responses mediated by brain presynaptic alpha2-adrenoceptors. In conclusion, these results do not support the hypothesis that chronic treatments with pilocarpine lead to a suitable model of alpha2-adrenoceptor supersensitivity.

Recovery of cardiac norepinephrine concentration and tyrosine hydroxylase activity by the central alpha2-adrenoceptor agonist guanabenz in rats with aortic constriction.

Depletion of cardiac norepinephrine has been reported in cardiac hypertrophy. This depletion causes less support for cardiac output in response to sympathetic nerve activation. The central nervous system is thought to be involved in this abnormality. Correction of this abnormality is expected to restore proper support for the heart. Clipping of the ascending aorta or a sham operation was performed in 10-week-old rats. At 4 weeks after the operation, the left ventricular norepinephrine concentration in clipped rats decreased (p<0.01). The clipped rats and sham-operated rats were treated with either guanabenz (1 mg/kg) or a vehicle for 4 weeks starting from fifth postoperative week. The level of left ventricular norepinephrine increased more in clipped rats treated with guanabenz (469+/-37 ng/g) than in clipped rats treated with a vehicle (325+/-28 ng/g). The norepinephrine concentration in the left ventricle recovered significantly after the treatment with guanabenz (p<0.001). Tyrosine hydroxylase activity in the left ventricle also recovered after treatment with guanabenz (p<0.01). Modulation of sympathetic nerve tone by the alpha2-adrenoceptor agonist restored cardiac norepinephrine concentration and tyrosine hydroxylase activity. This could be a new approach to the treatment of heart failure.

Ethanol selectively counteracts hypotension evoked by central I(1)-imidazoline but not alpha2-adrenergic receptor activation in spontaneously hypertensive rats.

Previous studies from our laboratory showed that ethanol counteracts hypotensive responses to clonidine in spontaneously hypertensive rats. This study investigated whether this effect of ethanol involves interaction with central alpha2-adrenoceptors or I(1)-imidazoline receptors or both. The effects of ethanol (0.5 or 1 g/kg, i.v.) or an equal volume of saline on hypotensive and bradycardic responses to clonidine (mixed alpha2-adrenoceptor/I(1)-imidazoline receptor agonist), rilmenidine (selective I(1)-imidazoline receptor agonist), or alpha-methylnorepinephrine (selective alpha2-adrenoceptor agonist) were studied in conscious spontaneously hypertensive rats. Intracisternal administration of clonidine (0.5 microg), rilmenidine (25 microg), or alpha-methylnorepinephrine (4 microg) elicited similar decreases in mean arterial pressure (MAP; 25-30 mm Hg) that lasted > or =60 min. Subsequent administration of ethanol (0.5 and 1 g/kg, i.v.) counteracted the hypotensive effect of clonidine in a dose-related manner. Ethanol (1 g/kg) increased the blood pressure to levels similar to baseline (preclonidine) levels, and blood pressure remained significantly (p < 0.05) higher compared with the corresponding values in saline-treated rats. Similarly, ethanol (0.5 and 1 g/kg, i.v.) dose-dependently counteracted the hypotensive effect of rilmenidine. The antagonizing effects of ethanol on hypotension evoked by clonidine and rilmenidine were comparable in terms of both magnitude and duration. In contrast, ethanol (0.5 or 1 g/kg) had no effect on hypotension evoked by alpha-methylnorepinephrine. Except for a brief increase in blood pressure by ethanol (1 g/kg) at 5 min, blood pressure values obtained in alpha-methylnorepinephrine-treated rats receiving any of the two doses of ethanol were similar to postsaline values. Ethanol had no effect on bradycardic responses to any of the three hypotensive agents. Blood ethanol concentrations were similar regardless of the antihypertensive drug used. We concluded that the adverse hemodynamic effect of ethanol on centrally mediated hypotensive responses depends on the types of receptors involved in the elicitation of this response. That ethanol counteracts decreases in blood pressure evoked by clonidine and rilmenidine but not by alpha-methylnorepinephrine suggests an interaction between ethanol and central pathways involved in I(1)-imidazoline receptor-mediated hypotension.

Characterisation of the cardiovascular pharmacology of medetomidine in the horse and sheep

Medetomidine was administered to sheep and horses at a dose rate of 5 Pg kg −1 (i.v.). Heart rate and blood pressure were recorded. Medetomidine induced bradycardia and a biphasic blood pressure response consisting of a transient hypertension followed by hypotension. Administration of prazosin (an alpha, adrenoceptor antagonist; 100μg kg 1, i.v.) had no effect on the cardiovascular response to medetomidine (5 Vg kg −1, i.v.), but inhibited the cardiovascular response of methoxamine (an alpha, adrenoceptor agonist; 75 pg kg −1, i.v.). L-659,066 (an alpha2 adrenoceptor antagonist which does not cross the blood brain barrier; 264 μg kg −, i.v.) attenuated the medetomidine induced bradycardia, but had no effect on the cardiovascular response to methoxamine. L-659,066 also reduced the medetomidine induced hypertension in sheep, but had less effect on the horse. It is concluded that both alpha 1 and alpha 2 adrenoceptors are important in the control of cardiovascular function in horses and sheep. Medetomidine appears to act on alpha2 adrenoceptors alone in the sheep. The cardiovascular effects of medetomidine in the horse are complex and may be influenced by central alpha2 adrenoceptor regulation or effects on other receptor subtypes as well as direct stimulation of peripheral alpha 2 adrenoceptors.

Peripherally administered alpha2-adrenoceptor agonist in the modulation of chronic allodynia induced by spinal nerve ligation in the rat.

We studied whether a peripherally administered alpha2-adrenoceptor agonist modulates mechanical allodynia caused by unilateral ligation of two spinal nerves in the rat. Medetomidine, an alpha2-adrenoceptor agonist, atipamezole, an alpha2-adrenoceptor antagonist, or saline (control) was administered into the footpad of either the allodynic or the contralateral hindpaw. Medetomidine (1-10 microg/kg in 50 microL) reversed the unilateral allodynia in a dose-dependent fashion independent of the site of administration. At this dose range, medetomidine did not influence the heat-induced tail-flick response. The antiallodynic effect of medetomidine was completely reversed by a dose of atipamezole that alone was ineffective (30 microg/kg). At the largest dose used (100 microg/kg in 50 microL), atipamezole decreased the latency of the heat-induced tail flick and had an ipsilateral allodynic effect in the injected paw when administered into the control side. The atipamezole-induced mechanical allodynia was not attenuated by medetomidine. The results indicate that an alpha2-adrenoceptor agonist at a subantinociceptive dose may significantly attenuate allodynia produced by spinal nerve ligation. This antiallodynic effect is not due to a peripheral action, but rather to action on central (presumably spinal) alpha2-adrenoceptors. The allodynic effect of a high concentration of atipamezole (100 microg/kg in 50 microL) in the injected paw can be explained by peripheral nonadrenergic mechanisms (e.g., local irritation). The present behavioral results indicate that a selective alpha2-adrenoceptor agonist at a subantinociceptive dose effectively attenuates mechanical allodynia induced by an experimental model of chronic neuropathy in the rat. This antiallodynic action can be explained by central, rather than peripheral, alpha2-adrenergic mechanisms.

A Comparative Study of the Antinociceptive Action of Xenon and Nitrous Oxide in Rats

We attempted to clarify the mechanism of antinociceptive action induced by xenon and nitrous oxide.Eighty percent of nitrous oxide or 80% xenon was applied to rats inside enclosed clear plastic glass cylinders with their tails protruding for assessment of the tail-flick response to radiant heat. With repeated testing, there was a rapid reduction to nitrous oxide antinociception within 90 min, which was interpreted as development of tolerance, but not to xenon antinociception. Nitrous oxide antinociception was blocked by the intraperitoneal administration of 0.1 or 1.0 mg/kg yohimbine, but not by 1.0 or 5.0 mg/kg L659-066 or by 5.0 or 10 mg/kg naloxone. Xenon antinociception was not affected by any of these drugs. Yohimbine and L659-066 are characterized as alpha2-adrenoceptor antagonists. Although yohimbine penetrates the blood-brain barrier after systemic administration, L659-066 does not penetrate it and act peripherally. Therefore, the results indicate that alpha2-adrenoceptors, but not opioid receptors, may play a key role in antinociception induced by nitrous oxide in the central nervous system. Furthermore, the mechanism of xenon antinociception differs from that of nitrous oxide because it does not involve either alpha2 or opioid receptors. Implications: The precise mechanism of antinociceptive action of nitrous oxide and xenon remains unknown. It is still controversial whether an opioid system plays a role in antinociception induced by nitrous oxide. The results of the study showed that antagonism of central alpha2-adrenoceptors, but not opioid receptors, reverses the antinociception induced by nitrous oxide but not by xenon, which indicates that alpha2-adrenoceptors may play a key role in nitrous oxide antinociception. (Anesth Analg 1997;85:931-6)

Inhibition of hyperalgesia through peripheral and central alpha2-adrenoceptor activation in the inflamed knee joint model of the rat

Recently, the location of peripherally distributed alpha2-adrenoceptors was demonstrated in a model of inflammatory pain [1]. The inflamed knee joint model of the rat presents a well-established pre-clinical model of thermal hyperalgesia. Inhibition of hyperalgesia could be achieved by either peripheral (intra-articular (IA) or central (intrathecal (IT)) delivery of clonidine. However, until now no data have existed, about the superiority of central or peripheral administration of clonidine for the inhibition of thermal hyperalgesia. As it was shown for peripheral opioid receptors the analgesic efficacy of peripherally acting clonidine could depend on the state of inflammation. Thus, in this present study we sought to evaluate the antinociceptive action of intra-articular and intrathecal administered clonidine with regard to time effect and dose-effect relation for two distinct routes of alpha2-agonist application. After IAR approval, male Sprague-Dawley rats received a chronic intrathecal catheter (i.c.). Inflammation was induced once by IA injection of 3% carrageenan-caolin (CC; 100 μL) into the right knee joint. Thermal hyperalgesia (TH) was tested using a modified Hargreaves device every 30 min up to 4 h on day 1 and day 2 (6-8 rats/group): 1=IT saline/IA saline; 2=sham-operated rats/IA saline; 3=IT clonidine/IA saline; 4=IT saline/IA clonidine. Drug delivery was 30 min after induction of inflammation. Thermal hyperalgesia of the hind paw was stable during the 28 h observation period (P<0.05). IT and IA clonidine resulted in a dose-dependent inhibition of TH, revealing similar ED50 on both days. All effects were reversible by IT or IA yohimbine. This study suggests a similar efficacy for inhibition of thermal hyperalgesia by peripheral and central administration of an alpha2-adreno-agonist such as clonidine for inflammatory pain. Unlike the response to opioids, this analgesic activity does not appear to depend on the duration of inflammation Fig. 3.Fig. 3: (abstract 41)

Centrally acting antihypertensives

Classic centrally acting antihypertensives are known to stimulate alpha2-adrenoceptors located in the pontomedullary region, in the vicinity of the nucleus tractus solitarii, vasomotor centre, vagal nucleus and the various interconnecting neurones. The stimulation of these central alpha2-adrenoceptors induces peripheral sympathoinhibition and hence a reduction in (elevated) blood pressure, predominantly as a result of vasodilation and a consequent decrease in peripheral vascular resistance. Clonidine, guanfacine, guanabenz and alpha-methyldopa (via its active metabolite alpha-methylnoradrenaline) are well-known examples of classic centrally acting antihypertensives. They are effective antihypertensives with an attractive haemodynamic profile. However, these agents have lost much of their clinical interest because of their subjectively unpleasant side-effects (sedation, dry mouth, impotence). Since these side-effects are also mediated, to a major extent, by alpha2-adrenoceptors it is virtually impossible to separate the desired centrally induced antihypertensive effect and the adverse reactions by designing new compounds. I1-Imidazoline receptors have recently been discovered as a new target of centrally acting antihypertensives. When stimulated with agonists the I1-imidazoline receptors, located in the nucleus reticularis lateralis will trigger peripheral sympathoinhibition, following similar pathways as involved in the effects of the classic alpha2-adrenoceptor stimulants. Moxonidine and rilmenidine are I1-imidazoline receptor stimulants with little affinity for alpha2-adrenoceptors. Accordingly, such agents lower elevated blood pressure in a similar manner as the aforementioned older drugs, but it may be hoped that their side-effect profile is more favourable. Accordingly, it would now be possible to separate the attractive haemodynamic properties and the side-effects of centrally acting antihypertensives.

Modulation of sympathetic outflow by centrally acting antihypertensive drugs.

The modulation of peripheral sympathetic activity by the central nervous system (CNS) has been intensely investigated as a potential target of antihypertensive drugs. In particular, clonidine, guanfacine, and alpha-methyl-DOPA (acting via its metabolite alpha-methylnoradrenaline) have been developed as antihypertensives with a predominantly CNS site of action. Initially these drugs have been assumed to reduce elevated blood pressure via the stimulation of central alpha2-adrenoceptors in the brain stem, thus leading to peripheral sympathoinhibition and a reduction of elevated blood pressure, heart rate, and plasma catecholamines. In a later stage it has been recognized that central imidazoline (I1) receptors, probably located in the nucleus reticularis lateralis in the medullary region, may also be the target of centrally acting antihypertensives. Moxonidine and rilmenidine are the prototypes of such agents. Accordingly, the receptor profile of the various types of centrally acting antihypertensives may be characterized as follows: alpha-methyl-DOPA, alpha2 (through alpha-methylnoradrenaline); clonidine alpha2 + I1 (mixed agonist); and moxonidine, rilmenidine, I1 > alpha2. The various compounds mentioned will thus cause peripheral sympathoinhibition, initiated by different receptor targets in the CNS. Finally, the peripheral alpha1-blocker, urapidil, has been demonstrated to possess an additional central mechanism, mediated by the stimulation of serotonergic 5HT1A-receptors located in the rostral ventrolateral medulla. The stimulation of these 5HT1A-receptors appears to suppress, via the autonomic nervous system, the reflex tachycardia triggered by vasodilation.

Cardiovascular responses to static exercise in conscious cats: effects of intracerebroventricular injection of clonidine.

1. Static exercise elicits increases in arterial blood pressure and heart rate (HR) in humans and conscious animals. In this study, the effects of intracerebroventricular (I.C.V.) administration of clonidine, an alpha 2-adrenergic agonist, on these cardiovascular responses were investigated using conscious cats. Four cats were operantly trained to extend a forelimb and press a bar (200-650 g) for 15-60 s. A stainless-steel cannula was inserted into the right lateral ventricle for I.C.V. injection of drugs, and a common carotid artery was catheterized to measure mean arterial pressure (MAP) and HR. The number of exercise trials and changes in MAP, HR and force were pooled for 30 min periods. After the cats exercised for 30 min, either artificial cerebrospinal fluid (CSF) or clonidine (2 or 5 micrograms) were administered intracerebroventricularly. 2. Before clonidine injection, fifty-two exercise trials increased MAP and HR by 15 +/- 3 mmHg and 41 +/- 5 beats min-1, respectively. Administration of clonidine (2 micrograms) did not alter the resting MAP and HR, but attenuated the increases in MAP and HR in response to exercise (0-30 min post-clonidine: n = 81; delta MAP, 6 +/- 3 mmHg; delta HR, 20 +/- 6 beats min-1; 30-60 min post-clonidine: n = 71; delta MAP, 4 +/- 4 mmHg; delta HR, 17 +/- 8 beats min-1). Administration of artificial CSF I.C.V. had no effect on the cardiovascular responses to static exercise. 3. An increased dose of clonidine (5 micrograms) decreased resting MAP and HR by 31 +/- 7 mmHg and 37 +/- 7 beats min-1, respectively, and markedly blunted the cardiovascular responses to exercise (pre-clonidine: n = 52; delta MAP, 17 +/- 3 mmHg; delta HR, 38 +/- 5 beats min-1; post-clonidine 0-30 min: n = 66; delta MAP, 4 +/- 2 mmHg; delta HR, 15 +/- 5 beats min-1; post-clonidine 30-60 min: n = 60; delta MAP, 4 +/- 2 mmHg; delta HR, 14 +/- 6 beats min-1). Pretreatment with the alpha 2-adrenergic antagonist, yohimbine (8 micrograms, I.C.V.), blocked the attenuating effects of I.C.V. administration of clonidine (5 micrograms). 4. These results show that stimulation of central alpha 2-adrenoceptors by clonidine attenuates the cardiovascular responses to static exercise in conscious cats. In addition, this study suggests that alpha 2-adrenoceptors blocked by yohimbine injected I.C.V. do not appear to have a tonic influence on HR and blood pressure.