Attentional performance of (C57BL/6Jx129Sv)F2 mice in the five-choice serial reaction time task.

Impaired attention is evident in several neurological and psychiatric disorders. In the present study, attentional capabilities were measured in the operant five-choice serial reaction time task (5-CSRTT) in male (C57BL/6Jx129Sv)F2 hybrid (B6129F2) mice. Main aims were to validate and standardize the test in these mice: to setup procedures, measure potential beneficial effects of sub-chronic nicotine in degraded versions of the 5-CSRTT (by decreasing stimulus duration, inducing white noise and making the stimuli unpredictable) and study disruptive effects of additional administration of the muscarinic antagonist scopolamine. During the baseline pre-nicotine sessions, the B6129F2 mice attained a very good performance in the test (95% accuracy). As stimulus duration was reduced from 2 s to 1 s, response accuracy of the mice decreased. Mice treated with nicotine (0.16 mg/kg) attained significantly higher response accuracy and had a lower percentage of incorrect responses in comparison with the solvent-treated animals. No further beneficial effects of nicotine were found. Reduced response accuracy was also obtained when stimulus duration was reduced from 1 s to 0.5 s and when a variable intertrial interval was introduced. Noise interpolation between trials did not impair performance. Finally, scopolamine (0.16 mg/kg) disrupted attentional functioning. Although most studies have been performed in rats, these results add to the existing evidence that the 5-CSRTT can also be used to assess attentional performance in mice. This offers the opportunity to test transgenic and knockout mice with similar background as the B6129F2 as animal models of psychiatric and neurological diseases.

Risperidone: a novel antipsychotic with balanced serotonin-dopamine antagonism, receptor occupancy profile, and pharmacologic activity.

The interaction of risperidone, 9-hydroxyrisperidone (the principal active metabolite), and clozapine with neurotransmitter receptors was investigated in vitro using animal brain tissue homogenates and cloned human receptors expressed in cells and ex vivo using quantitative receptor autoradiography in rat and guinea pig brain sections. In vitro, risperidone and 9-hydroxyrisperidone had similar binding profiles, and their highest affinity was for 5-HT2A receptors (cloned human, Ki 0.4 nM); affinities for other 5-HT-receptor subtypes were at least 100 times lower. Risperidone bound to 5-HT2A receptors with 20 times greater affinity than clozapine and 170 times greater affinity than haloperidol. Clozapine primarily bound to histamine H1 receptors and haloperidol to dopamine D2 receptors. The binding affinity of risperidone and 9-hydroxyrisperidone for the D2 family of receptors (D2L, D2S, D3, D4) was one order of magnitude lower than their affinity for 5-HT2A receptors. Risperidone bound to D2 and D3 receptors with 50 and 20 times greater affinity than clozapine and was only 2 to 3 times less potent than haloperidol. All compounds bound with similar affinities to D4 receptors (Ki 5-9 nM), and their affinities for D1 receptors were 100 times lower than for D4 receptors. The ex vivo receptor occupancy profile of the compounds matched the in vitro receptor binding profile. A conspicuous property of risperidone, not seen for the other compounds, was the shallow occupancy curve at D2 receptors in the striatum and mesolimbic brain area. Moreover, it was observed that antagonism of strong D2-receptor stimulation by apomorphine in rats was achieved at less than 50% D2 occupancy by the antipsychotics.(ABSTRACT TRUNCATED AT 250 WORDS)

Differential effects of the new antipsychotic risperidone on large and small motor movements in rats: a comparison with haloperidol.

Risperidone, a new antipsychotic agent, was studied for its effects on spontaneous motor activity in rats in comparison with haloperidol. Motor activity was recorded via the optical scanning technique (horizontal and vertical activity) and via a recently developed technique based on the piezo-electric principle which, in contrast to optical scanning, is very sensitive to small, stationary movements (piezo activity). Risperidone and haloperidol at low doses depressed both vertical activity (ED50s: 0.062 and 0.038 mg/kg, respectively) and horizontal activity (0.18 and 0.060 mg/kg, respectively). With increase of dose, motor activity decline was significantly faster with haloperidol than with risperidone. Moreover, haloperidol also rapidly depressed piezo activity (ED50: 0.085 mg/kg) whereas risperidone depressed this component of motor behaviour at much higher doses only (ED50: 2.80 mg/kg). Visual inspection did not reveal abnormal behavioural movements following the test compounds. Risperidone, therefore, preserves normal small movements over a much larger dose interval than haloperidol; this effect may be related to its relatively low cataleptogenic activity and potentially also to a reduced EPS liability. The present results further confirm that the piezo technique may complement the optical scanning method, and thereby enhance the information on the extent that test compounds modify behaviour.

Behavioral activity of rats measured by a new method based on the piezo-electric principle.

Spontaneous and drug-induced (haloperidol, apomorphine, and amphetamine) motor activity of rats was measured simultaneously via two distinct and independent methods: the classical optical scanning technique and a new procedure based on the piezo-electric principle. The latter procedure measured animal-induced mechanical vibrations of a flexible cage floor which were transduced into electric signals via piezo-electricity. The piezo method appeared to be relatively more sensitive in recording the small, stereotyped motor movements induced by apomorphine (0.63- greater than or equal to 10 mg/kg) and high doses of amphetamine (2.5- greater than or equal to 20 mg/kg). The optical scanning technique, on the other hand, was more sensitive in recording horizontal displacements across the cage such as induced by low doses of amphetamine (0.31-2.5 mg/kg). Both methods showed comparable sensitivity in recording the depression of behaviour induced by haloperidol (0.04- greater than or equal to 1.25 mg/kg) or low doses of apomorphine (0.04-0.16 mg/kg). The piezo method may complement the optical scanning procedure, and thereby enhance the information on the extent that test compounds modify animal behaviour.

Interaction of antipsychotic drugs with neurotransmitter receptor sites in vitro and in vivo in relation to pharmacological and clinical effects: role of 5HT2 receptors.

In the introductory section an overview is given of the strategies which have been proposed in the search for side-effect free antipsychotics. Special attention is paid to the role of predominant 5HT2 receptor blockade over D2 blockade. Whereas D2 receptor blockade seems to be essential for the treatment of positive symptoms of schizophrenia, it also underlies the induction of extrapyramidal side effects (EPS). Predominant 5HT2 receptor blockade may reduce the EPS liability and can ameliorate negative symptoms of schizophrenia. We further report a nearly complete list of neuroleptics that are on the European market and eight new antipsychotics that recently entered clinical trial, 5HT2 and D2 receptor binding affinity (Ki values) and the rank order in affinity for various neurotransmitter receptor subtypes are also discussed. For the eight new antipsychotics and for six reference compounds the complete receptor binding profile (including 33 radioligand receptor binding and neurotransmitter uptake models) is reported. Furthermore, for a series of 120 compounds the relative affinity for D2 receptors and D3 receptors (a recently cloned new dopamine receptor subtype) is compared. Finally, original findings are reported for the new antipsychotic risperidone and for haloperidol and clozapine on the in vivo occupation of neurotransmitter receptors in various brain areas after systemic treatment of rats or guinea pigs. The receptor occupation by the drugs was measured ex vivo by quantitative receptor autoradiography. The receptor occupancy was related to the motor activity effects of the test compounds (measurements were done in the same animals) and to the ability of the drugs to antagonize various 5HT2 and D2 receptor mediated effects. With risperidone a high degree of central 5HT2 receptor occupation was achieved before other neurotransmitter receptors became occupied. This probably co-underlies the beneficial clinical properties of the drug. Antagonism of the various D2 receptor-mediated effects was achieved at widely varying degrees of D2 receptor occupancy, from just about 10% to more than 70%. For therapeutic application it may be of prime importance to carefully titrate drug dosages. Antipsychotic effects may be achieved at a relatively low degree of D2 receptor occupancy at which motor disturbances are still minimal. With drugs such as risperidone that produce shallow log dose-effect curves, differentiation between the various D2 receptor mediated effects may be made more easily, allowing EPS-free maintenance therapy of schizophrenic patients.

Regional brain distribution of risperidone and its active metabolite 9-hydroxy-risperidone in the rat.

Risperidone is a new benzisoxazole antipsychotic. 9-Hydroxy-risperidone is the major plasma metabolite of risperidone. The pharmacological properties of 9-hydroxy-risperidone were studied and appeared to be comparable to those of risperidone itself, both in respect of the profile of interactions with various neurotransmitters and its potency, activity, and onset and duration of action. The absorption, plasma levels and regional brain distribution of risperidone, metabolically formed 9-hydroxy-risperidone and total radioactivity were studied in the male Wistar rat after single subcutaneous administration of radiolabelled risperidone at 0.02 mg/kg. Concentrations were determined by HPLC separation, and off-line determination of the radioactivity with liquid scintillation counting. Risperidone was well absorbed. Maximum plasma concentrations were reached at 0.5-1 h after subcutaneous administration. Plasma concentrations of 9-hydroxy-risperidone were higher than those of risperidone from 2h after dosing. In plasma, the apparent elimination half-life of risperidone was 1.0 h, and mean residence times were 1.5 h for risperidone and 2.5 h for its 9-hydroxy metabolite. Plasma levels of the radioactivity increased dose proportionally between 0.02 and 1.3 mg/kg. Risperidone was rapidly distributed to brain tissues. The elimination of the radioactivity from the frontal cortex and striatum--brain regions with high concentrations of 5-HT2 or dopamine-D2 receptors--became more gradual with decreasing dose levels. After a subcutaneous dose of 0.02 mg/kg, the ED50 for central 5-HT2 antagonism in male rats, half-lives in frontal cortex and striatum were 3-4 h for risperidone, whereas mean residence times were 4-6 h for risperidone and about 12 h for 9-hydroxy-risperidone. These half-lives and mean residence times were 3-5 times longer than in plasma and in cerebellum, a region with very low concentrations of 5-HT2 and D2 receptors. Frontal cortex and striatum to plasma concentration ratios increased during the experiment. The distribution of 9-hydroxy-risperidone to the different brain regions, including frontal cortex and striatum, was more limited than that of risperidone itself. This indicated that 9-hydroxy-risperidone contributes to the in vivo activity of risperidone, but to a smaller extent than would be predicted from plasma levels. AUCs of both active compounds in frontal cortex and striatum were 10-18 times higher than those in cerebellum. No retention of metabolites other than 9-hydroxy-risperidone was observed in any of the brain regions investigated.

Survey on the pharmacodynamics of the new antipsychotic risperidone.

This review reports on the pharmacodynamics of the new antipsychotic risperidone. The primary action of risperidone is serotonin 5-HT2 receptor blockade as shown by displacement of radioligand binding (Ki: 0.16 nM), activity on isolated tissues (EC50: 0.5 nM), and antagonism of peripherally (ED50: 0.0011 mg/kg) and centrally (ED50: 0.014 mg/kg) acting 5-HT2 receptor agonists in rats. Risperidone is at least as potent as the specific 5-HT2 receptor antagonist ritanserin in these tests. Risperidone is also a potent dopamine D2 receptor antagonist as indicated by displacement of radioligand binding (Ki: 1.4 nM), activity in isolated striatal slices (IC50: 0.89 nM), and antagonism of peripherally (ED50: 0.0057 mg/kg in dogs) and centrally acting D2 receptor agonists (ED50: 0.056-0.15 mg/kg in rats). Risperidone shows all effects common to D2 antagonists, including enhancement of prolactin release. However, some central effects such as catalepsy and blockade of motor activity occur at high doses only. Risperidone is 4-10 times less potent than haloperidol as a central D2 antagonist in rats and it differs from haloperidol by the following characteristics: predominant 5-HT2 antagonism; LSD antagonism; effects on sleep; smooth dose-response curves for D2 antagonism; synergism of combined 5-HT2/D2 antagonism; pronounced effects on amphetamine-induced oxygen consumption; increased social interaction; and pronounced effects on dopamine (DA) turnover. Risperidone displays similar activity at pre- and postsynaptic D2 receptors and at D2 receptors from various rat brain regions. The binding affinity for D4 and D3 receptors is 5 and 9 times weaker, respectively, than for D2 receptors; interaction with D1 receptors occurs only at very high concentrations. The pharmacological profile of risperidone includes interaction with histamine H1 and alpha-adrenergic receptors but the compound is devoid of significant interaction with cholinergic and a variety of other types of receptors. Risperidone has excellent oral activity, a rapid onset, and a 24-h duration of action. Its major metabolite, 9-hydroxyrisperidone, closely mimics risperidone in pharmacodynamics. Risperidone can be characterized as a potent D2 antagonist with predominant 5HT2 antagonistic activity and optimal pharmacokinetic properties.

Occupancy of central neurotransmitter receptors by risperidone, clozapine and haloperidol, measured ex vivo by quantitative autoradiography.

Risperidone (Risperdal) is a novel antipsychotic drug, with beneficial effects on both positive and negative symptoms of schizophrenia, and with a low incidence of extrapyramidal side effects (EPS). These particular properties have been attributed to the predominant and very potent serotonin 5-HT2 receptor antagonism of the drug combined with less potent dopamine D2 antagonism. In order to provide data on the degree to which various central neurotransmitter receptors are occupied in vivo, we performed ex vivo receptor occupancy studies with risperidone in comparison with clozapine and haloperidol in rats and guinea pigs. Various types of receptors, to which the compounds were known to bind to in vitro, were investigated precisely using receptor autoradiography in sections of the same rat brain except for histamine H1 receptors that were measured in the guinea-pig cerebellum. Risperidone (2 h after s.c. treatment) occupied 5-HT2 receptors at very low doses (ED50 = 0.067 mg/kg). Nearly full occupancy (> 80%) was achieved before H1, D2, alpha 1 and alpha 2 receptors became occupied (ED50 = 0.45, 0.66, 0.75 and 3.7 mg/kg, respectively). Clozapine displayed occupancy of H1 and alpha 1 receptors at low doses (ED50 = 0.15 and 0.58 mg/kg, respectively) and of 5-HT2, 5-HT1C, D2, alpha 2, cholinergic muscarinic and 5-HT1A receptors at higher doses (ED50 = 1.3, 1.8, 9.0, 9.5, 11 and 15 mg/kg, respectively). Haloperidol occupied D2 and alpha 1 receptors at low doses (ED50 = 0.13 and 0.42 mg/kg, respectively) and 5-HT2 receptors at a higher dose (ED50 = 2.6 mg/kg). Occupancy of receptor types occurred with similar ED50-values in various brain areas, e.g. D2 receptors in striatum and mesolimbic areas. The ED50-values for the ex vivo measured occupancy of 5-HT2 and D2 receptors were in good agreement with ED50-values for functional effects putatively mediated by these central receptors. The dose-dependent occupancy of D2 receptors proceeded more gradually with risperidone (slope in the caudate-putamen: 0.85) than with clozapine (slope: 1.44) or haloperidol (slope: 1.51). It has previously been suggested that partial D2 receptor occupancy may suffice to control the positive symptoms of schizophrenia, whereas higher D2 receptor occupancy would induce extrapyramidal symptoms (EPS). The dose ratio for high (75%) vs. low (25%) D2 receptor occupancy in the caudate-putamen, was 37.3 for risperidone, 8.4 for clozapine, and 7.9 for haloperidol.(ABSTRACT TRUNCATED AT 400 WORDS)

Presence of a particular subpopulation of dopamine receptors within the septal nuclei: a behavioural study on cats.

The behavioural effects of intraseptal administration of dopaminergic drugs (apomorphine, haloperidol, (3,4-dihydroxy-phenylamino)-2-imidazoline (DPI), ergometrine and dopamine) and alpha-noradrenergic drugs (oxymetazoline, noradrenaline and phentolamine) were analysed in cats pretreated with morphine (5 mg/kg, i.p.). Changes in frequencies of stereotyped locomotor patterns were used for statistical evaluation of drug-induced effects. Taking advantage of the specificity of the drugs mentioned, a distinction could be made between effects mediated via excitation-mediating dopamine (DAe), inhibition-mediating dopamine (DAi) and alpha-noradrenaline (alpha-NA) receptors. Intraseptal injection of the DAi agonist DPI resulted in a decrease in the frequency of stereotyped locomotor patterns. This effect was dose-dependent and mimicked by that of intraseptally applied dopamine but not of any of the other drugs. Moreover, intraseptal injection of the DAi antagonist ergometrine inhibited the effect of DPI. The DAe agonist apomorphine as well as the DAe antagonist haloperidol remained ineffective when applied in low doses. The alpha-NA antagonist phentolamine and a rather high dose of haloperidol produced a slight but significant increase in the frequency of locomotor patterns; intraseptally applied oxymetazoline counteracted the phentolamine-induced effect. It is concluded that the septal nuclei of cats contain functionally active, alpha-NA receptors as well as functionally active, dopamine (DA) receptors having pharmacological properties identical to those of DA receptors present within the mesolimbic structures such as the nucleus accumbens: the so-called inhibition-mediating DAi receptors.

Further validation of in vivo and in vitro pharmacological procedures for assessing the alpha 2/alpha 1-selectivity of test compounds: (1). Alpha-adrenoceptor antagonists.

Twenty one chemically dissimilar alpha-adrenoceptor antagonists were assessed in 7 different tests. The potencies of the compounds to protect rats from norepinephrine lethality were closely correlated with their potencies for inhibition of [3H]WB4101, but not of [3H]clonidine, [3H]idazoxan, [3H]rauwolscine or [3H]yohimbine binding to rat brain homogenates. The reverse was found for the potencies of the compounds to prevent the antidiarrheal effect of clonidine. The in vivo anti-norepinephrine/anti-clonidine potency ratios were highly correlated with the ratios between the in vitro potency to inhibit [3H]WB4101 binding on one hand and [3H]clonidine, [3H]idazoxan, [3H]rauwolscine or [3H]yohimbine binding on the other. The reliability of the above models for measuring alpha 1- and alpha 2-adrenoceptor antagonistic activity and selectivity is discussed.

Further validation of in vivo and in vitro pharmacological procedures for assessing the alpha 2/alpha 1-selectivity of test compounds: (2). Alpha-adrenoceptor agonists.

Eighteen chemically dissimilar alpha-adrenoceptor agonists were assessed in 6 different tests. The antidiarrheal activity of the compounds was highly correlated with the diuretic activity. Both activities were highly correlated with the inhibition of [3H]clonidine or [3H]idazoxan binding; the degree of correlation with inhibition of [3H]WB4101 binding or with antiptotic activity was much less. Antiptotic activity was better correlated, although still poorly, with inhibition of [3H]WB4101 binding than with inhibition of [3H]clonidine or [3H]idazoxan binding. The in vivo antiptotic/diuretic or antiptotic/antidiarrheal potency ratios both reflected the known alpha 2/alpha 1-selectivity of the compounds tested and were significantly correlated with the in vitro potency ratios between inhibition of [3H]WB4101 binding on one hand and of [3H]clonidine or [3H]idazoxan binding on the other. The reliability of the above models for measuring alpha 1- and alpha 2-adrenoceptor agonistic activity and selectivity is discussed.

Normalization of small intestinal propulsion with loperamide-like antidiarrheals in rats.

Gastrointestinal propulsion and the presence of diarrhea were assessed in rats pretreated with various opioids and challenged orally with either castor or paraffin oil, which both contained phenol red as a marker of gastrointestinal propulsion. In solvent-pretreated rats, diarrhea was always observed within 90 min after castor oil, reflecting a state of hyperpropulsive activity of the gut, but never (up to 8 h) after paraffin oil, reflecting normal intestinal propulsion (which amounted to an average distance of 91% of the total length of the small intestine in 90 min). Paraffin oil propulsion was blocked (to values less than 60%) by all opioids tested with the exception of the gut-selective compounds loperamide, loperamide oxide and fluperamide oxide (ED50s: greater than or equal to 160 mg/kg). Castor oil diarrhea was antagonized by all opioids tested and, at comparable but slightly (1.3-2.6 times) higher doses, propulsion was normalized to values (less than 100%) comparable to those measured in paraffin oil-challenged control rats. Castor oil propulsion was further reduced to subnormal values (less than 60%) by still higher doses of the opioids, comparable to those that blocked propulsion after paraffin oil. However, the required dose increment varied consistently among the opioids tested and ranged, depending on gut selectivity, from a factor 2.3 times the antidiarrheal dose for narcotic analgesics such as pethidine and dextromoramide to greater than 300 for antidiarrheals such as loperamide, loperamide oxide and fluperamide oxide. Protection from diarrhea and normalization of propulsion showed a close correlation; both failed to correlate with central analgesic activity and are thought to be mediated via peripheral opioid receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of the analgesic and intestinal effects of fentanyl and morphine in rats.

Clinical studies report a low incidence of intestinal side effects with transdermally administered fentanyl (TTS-fentanyl) in comparison with oral morphine. To support these clinical data, analgesic and intestinal effects of both opioids were compared in rats. After subcutaneous injection, analgesia in the tail withdrawal reaction test was obtained at a peak effect dose of 0.032 mg/kg with fentanyl and 8.0 mg/kg with morphine. This analgesic dose exceeded the ED50 for inhibition of castor oil-induced diarrhea only slightly (1.1 x) in the case of fentanyl (0.028 mg/kg) but markedly (36 x) in the case of morphine (0.22 mg/kg). To reverse completely the antidiarrheal effect of equivalent analgesic doses of the opioids (their ED50S for analgesia lasting 2 hours), much more naloxone was required in the case of morphine (5.4 mg/kg) than in the case of fentanyl (0.19 mg/kg). After oral administration, the difference between both opioids was less pronounced. Analgesia was obtained at 0.85 mg/kg with fentanyl and 32 mg/kg with morphine. This analgesic dose only slightly (1.7 x) exceeded the antidiarrheal dose in the case of fentanyl (0.49 mg/kg) but significantly (6.2 x) in the case of morphine (5.2 mg/ kg). To reverse completely the antidiarrheal effect of equivalent analgesic oral doses of the opioids (their ED50S for analgesia lasting 2 hours), more naloxone was required in the case of morphine (11 mg/kg) than in the case of fentanyl (2.0 mg/kg). Rapid penetration of fentanyl into the brain is thought to be responsible for small dissociation between the analgesic and intestinal effect of this lipophilic opioid. The present data provide preclinical evidence to support the relatively low incidence of intestinal side effects observed clinically with the use of TTS-fentanyl in comparison with orally administered morphine.

Involvement of the septum in central dopamine-acetylcholine interactions in morphine-treated cats.

The involvement of the septum in central dopamine-acetylcholine (DA-ACh) interactions was investigated by analysis of the behavioural effects of intracerebrally injected drugs in cats pretreated with morphine (5 mg/kg, IP). The intracerebrally evoked effects on the morphine-induced behaviour were analyzed both quantitatively (changes in the incidence of locomotor patterns) and qualitatively (changes in the stereotyped nature of the behaviour patterns). Activation of a particular subclass of dopamine receptors (DAi receptors) within the septum by means of the DAi agonist (3,4-dihydroxyphenylamino)-2-imidazoline (DPI) suppressed the effect of intraseptal injections of the cholinergic agonist carbachol. Simultaneous activation of the DAi receptors within the septum and those within the caudate nucleus produced an effect characteristics of activation of septal DAi receptors; the effect characteristic of activation of caudate DAi receptors was suppressed. Analogous results were obtained when the DAi receptors within the septum and those within the caudate nucleus were simultaneously inhibited by the DAI antagonist ergometrine. Finally, it was found that the effect of intraseptal injections of the cholinergic agonist carbachol was suppressed by inhibition of the DAi receptors within the caudate nucleus. In the latter case, the effect characteristic of inhibition of caudate DAi receptors was also suppressed. The present results indicate the existence of interactions between (1) septal ACh activity and septal DAi activity, (2) septal DAi activity and caudate DAi activity, and (3) septal ACh activity . A mechanism of action for the observed interactions is discussed.

Intraseptally injected opiate agents: effects on morphine-induced behaviour of cats.

Behavioural effects of intraseptally administered opiate agents were analyzed in cats pretreated with an intraperitoneal injection of morphine. In this way, it became possible to investigate (1) the involvement of septal opiate receptors in the behavioural response of cats to systemic administration of morphine, and (2) the pharmacological character of septal opiate receptors. The following results were obtained with intraseptal injections 15-16 min after intraperitoneal morphine: (1) naloxone decreased frequencies of head and limb movements, and (2) morphine was ineffective. The following results were obtained with intraseptal injections 40-41 min after intraperitoneal morphine: (1) beta-endorphin and, to a lesser extent, fentanyl increased frequencies of locomotor patterns, (2) morphine and Met-enkephalin were ineffective, (3) naloxone and naltrexone decreased frequencies of locomotor patterns in a dose-dependent way, (4) naloxone and naltrexone antagonized the effects of beta-endorphin and fentanyl, and (5) morphine did not attenuate the effect of naloxone. The intraseptal injections affected only the frequencies of the systemically evoked behaviour patterns; the nature of the behaviour patterns remained unchanged. It is concluded that (1) systemically administered morphine does not affect behaviour via a direct action on septal opiate receptors, and (2) the receptors mediating the septally evoked effects are most probably epsilon-type opiate receptors. The hypothesis is put forward that systemic administration of morphine results in an increased release of beta-endorphin from hypothalamo-septal neurons and, as a consequence, changes the beta-endorphin activity at the epsilon-type opiate receptors in the septum.

Antagonism of the antidiarrhoeal effect of clonidine and the lethal effect of noradrenaline in rats: a reliable procedure to evaluate the in-vivo alpha 1- and alpha 2-blocking activity of drugs?

Eight compounds with alpha-adrenergic blocking activity were tested for their ability to antagonize the antidiarrhoeal effect of clonidine (clonidine test) and the lethal effect of noradrenaline (noradrenaline test). Six of the compounds studied are alpha-adrenergic blocking agents with known alpha 2/alpha 1 selectivity. Two compounds, ketanserin (R 41 468) and butanserin (R 53 393), are 5-hydroxytryptamine S2-antagonists. The ED50-values (mg kg-1) obtained in the clonidine test were: phentolamine (0.34), RX781094 (0.34), yohimbine (0.51), piperoxan (9.36), butanserin (greater than 5.0), prazosin (greater than 10.0), phenoxybenzamine (greater than 40.0), and ketanserin (greater than 80.0). In the noradrenaline test the ED50's (mg kg-1) were: butanserin (0.014), prazosin (0.032), phentolamine (0.59), phenoxybenzamine (1.02), ketanserin (4.69), RX781094 (12.4), piperoxan (21.5), and yohimbine (25.0). The selectivity alpha 2/alpha 1-ratios (ED50 clonidine/ED50 noradrenaline were: yohimbine (0.020), RX781094 (0.027), piperoxan (0.44), phentolamine (0.58), ketanserin (greater than 39), prazosin (greater than 312), and butanserin (greater than 357). These results show that yohimbine and RX781094 are equipotent and relatively selective alpha 2-antagonists; piperoxan and phentolamine block both alpha 1- and alpha 2-receptors at closely related doses; ketanserin, prazosin and butanserin are selective blockers of alpha 1-receptors, ketanserin being very weak, prazosin and butanserin being very potent compounds in this respect. The potent and selective alpha 1-blocking activity of butanserin, combined to its 5-HT S2-antagonism makes butanserin a very interesting experimental drug in view of earlier reported data concerning the amplifying effects between 5-hydroxytryptaminergic and noradrenergic vascular mechanisms.