Comparison of post-junctional α-adrenoceptors in iris dilator muscle of humans, and albino and pigmented rabbits

The relative potency of -adrenoceptor agonists and the dissociation constants of competitive antagonists were studied to characterize the post junctional -adrenoceptor of the human iris dilator muscle. The data obtained from human iris dilator tissue was compared to that from rabbit. The iris dilator muscle was mounted in an organ bath and tension changes were recorded. (–)-Norepinephrine, (–)-phenylephrine (PE), oxymetazoline and p-aminoclonidine caused contractile responses in albino rabbit, pigmented rabbit and human iris dilator muscle in a concentration-dependent manner. The imidazoline molecules were partial agonists. In rabbit iris dilator, desensitization occurred to repeated oxymetazoline application at an interval of 1 h but recovery to the agonist activity was complete in about 3 h. Exposure to cocaine (10 mol/l), hydrocortisone (100 mol/l) and U-0521, a catechol-O-methyltransferase inhibitor (100 pmol/l), significantly potentiated the response to norepinephrine by 92-, 32- and 7 fold in iris dilator tissue of albino rabbit, pigmented rabbit and human, respectively. After block of uptake₁ and uptake₂, the EC₅₀ values of norepinephrine in the albino rabbit, pigmented rabbit and human iris dilator did not differ and ranged from 99 to 195 nmol/l. Small but significant potentiation by uptake blockers was also observed in the responses to PE in the albino rabbit or pigmented rabbit iris dilator. The average maximum tension induced by 100 mol/l PE was 96 ± 11 mg (n = 10), 197 ± 11 mg (n = 11), 45 ± 5 mg (n = 27) in albino rabbit, pigmented rabbit and human iris dilator, respectively.In human iris dilator, the responses to PE were competitively antagonized by prazosin, 5-methylurapidil and phentolamine with apparent pKB values of 7.3, 6.6 and 7.5, respectively. The pK _B values of the prazosin-PE interaction in iris dilator of albino and pigmented rabbit were 8.6 and 6.4, respectively.These results suggest that the post-junctional -adrenoceptors in iris dilator may be similar to that in pigmented rabbit iris. The -adrenoceptor of the human or pigmented rabbit iris dilator may be characterized as _1L-adrenoceptor subtype. The -adrenoceptor of albino rabbit iris dilator appears to be a high affinity subtype.Furthermore, albino rabbit may not be the best strain for the drug research which is relevant to human ocular therapeutics.     

Agonist pharmacology at recombinant α1A- and α1L-adrenoceptors and in lower urinary tract α1-adrenoceptors

BACKGROUND AND PURPOSE

Two distinct α₁-adrenoceptor phenotypes (α_1A and α_1L) have recently been demonstrated to originate from a single α_1A-adrenoceptor gene. Here, we examined the agonist profiles of recombinant α_1A and α_1L phenotypes and of lower urinary tract (LUT) α₁-adrenoceptors.

EXPERIMENTAL APPROACH

A series of drugs (A61603, Ro 115–1240, NS-49, MK017 and ESR1150) originally developed for stress urinary incontinence (SUI) therapy were used to stimulate recombinant α_1A- and α_1L-adrenoceptor phenotypes, and their potencies and intrinsic activity estimated from Ca²⁺ responses. Agonist-induced contractions were also examined in LUT tissues of rats and humans and in human mesenteric artery and rat tail artery.

KEY RESULTS

All the drugs were potent agonists of the α_1A-adrenoceptor compared with the α_1L-adrenoceptor phenotype. Among them, Ro 115–1240 was shown to be an α_1A-specific partial agonist that produced partial contractions through α_1A-adrenoceptors in rat prostate and tail artery, but not in the other LUT tissues and human mesenteric artery. In contrast, P-come 102 showed full agonist activity at α_1A- and α_1L-adrenoceptors, but was less selective than noradrenaline for α_1A-adrenoceptors. Like noradrenaline, P-come 102 was highly potent at inducing contractions in all of the LUT tissues tested. However, the potency and intrinsic activity of P-come 102 were significantly lower than those of noradrenaline in human mesenteric artery.

CONCLUSIONS AND IMPLICATIONS

The α_1A- and α_1L-adrenoceptor phenotypes and LUT α₁-adrenoceptors were demonstrated to have distinct agonist profiles. As adrenergic contractions in LUT are predominantly mediated through α_1L-adrenoceptors, the development of α_1L-selective agonists may provide clinically useful drugs for SUI therapy.     

Actions of tizanidine on α1- and α2-adrenoceptors in the peripheral tissues

• 1.1. Tizanidine behaved as the partial agonist on the α₁-adrenoceptor in high doses (10⁻⁶−10⁻⁴ M) and as the α₂-adrenoceptor agonist in low doses (3 × 10⁻⁹−10⁻⁶ M).
• 2.2. Tizanidine is about one-third as potent as clonidine in α₂-agonistic effects.

     

Subtypes of α1-adrenoceptors in hippocampus of pigs,guinea-pigs,calves and humans: regional differences

Radioligand binding studies were performed with membranes of guinea-pig, pig, calf and human hippocampus using [¹²⁵I]BE 2254 (also known as [¹²⁵I]HEAT) as the radioligand. [¹²⁵I]BE 2254 bound with similar high affinity to saturable populations of recognition sites in all four membrane preparations. Competition curves obtained with a variety of ligangs (e.g., WB 4101, benoxathian, 5-methyl-urapidil) were biphasic and the profiles of the high- and low-affinity components of [¹²⁵I]BE 2254 binding were similar in all four membrane preparations. The data suggest that [¹²⁵I]BE 2254 labels two subtypes of α₁-adrenoceptors in the hippocampus of these species. [³H]WB 4101 was used to label α_1A recognition sites in pig hippocampus membranes. [³H]WB 4101 recognized with high affinity an apparently homogeneous class of sites, as suggested by monophasic saturation and competition experiments. The rank order of affinity of the compounds for the high-affinity component of [¹²⁵I]BE 2254 binding was similar to the rank order of affinity of these drugs for [³H]WB 4101 sites. The pharmacological profile of the low-affinity component of [¹²⁵I]BE 2254 binding was similar to that described recently for the α_1B-adrenoceptor cloned from DDT₁ cells. In autoradiographic studies with human hippocampal slices, CEC (chloroethylclonidine), an alkylating agent described to show selectivity for α_1B-adrenoceptors, displaced preferentially [¹²⁵I]BE 2254 binding from the molecular layer of the dentate gyrus. In contrast, WB 4101 an α_1A-adrenoceptor-selective ligand, displaced preferentially [¹²⁵I]BE 2254 binding in the hilus and the CA3 region. The data show that 2 subtypes of α₁-adrenergic recognition sites can be identified in the hippocampus. In the human hippocampus, α_1A sites are predominant in the hilus and the CA3 region, whereas α_1B sites are predominant in the molecular layer of the dentate gyrus. These subtypes show a similar pharmacological profile in man, calf and guinea-pig, and may have a different functional role in these two areas of the hippocampus.     

Autoreceptors and α2-adrenoceptors at the serotonergic axons of rabbit brain cortex

Summary Slices of the rabbit occipito-parietal cortex were preincubated with ³H-serotonin and then superfused and stimulated electrically (2 min at 3 Hz). In the absence of drugs, the stimulation-evoked overflow of tritium was approximately 3% of the tritium content of the tissue. Unlabelled serotonin and 5-carboxamido-tryptamine, when administered in the presence of 6-nitroquipazine, reduced the evoked overflow of tritium. Their effects were antagonized by metitepin (apparent pA₂ value 8.1) and (±)-cyanopindolol (apparent pA₂ value 6.4). Metitepin, but not cyanopindolol, increased evoked tritium overflow; the effect of metitepin was greater in the presence than in the absence of nitroquipazine. The evoked overflow of tritium was also depressed by clonidine, an effect antagonized by idazoxan (apparent pA₂ value 7.0) but not by prazosin. Phenylephrine caused a decrease only at high concentrations that simultaneously accelerated basal tritium efflux. Prazosin and idazoxan did not change evoked tritium overflow, and phentolamine increased it significantly only when administered in the presence of (+)-oxaprotiline. Rauwolscine produced an inhibition that was prevented by metitepin. It is concluded that the serotonergic axons of the rabbit occipitoparietal cortex possess presynaptic, release-inhibiting serotonin autoreceptors and ₂-adrenoceptors. The receptors appear to receive an input of endogenous serotonin and, to a lesser extent, noradrenaline, under the conditions of these in vitro experiments.     

Comparison between thermoregulatory effects mediated by α1- and α2-adrenoceptors in normothermic and febrile rabbits

• 1.1. In order to better delineate the profile of thermoregulatory action of α₁-adrenoceptor antagonists; corynanthine (CRN) and BMY 20064 (BMY), and α₂-adrenoceptor agonist; medetomidine (MDT) and B-HT 920 (BHT), the effect of intravenous administration of two doses of these drugs on rectal (T_re) and ear skin (T_e) temperatures, metabolic rate (M), respiratory evaporative heat loss (E_res) and respiratory rate (R_r) were examined in febrile and non-febrile rabbits.
• 2.2. Results indicate that α₁-adrenoceptor antagonists as well as α₂-adrenoceptor agonists markedly lowered body temperature exhibiting antipyretic and hypothermic actions. The hypothermic and antipyretic effect after the CRN or BMY, and BHT or MDT, treatment was associated with inhibition of metabolic rate and/or with body heat redistributed to peripheral tissues and an increase of the potential for heat loss to the environment.
• 3.3. BMY also abolished the thermogenic response to cold. However, BMY did not affect metabolic heat production on exposure to a cold ambient temperature. This unexpected phenomenon is difficult to explain at the present moment. Possible mechanisms responsible for the thermoregulatory activity of BMY are discussed.
• 4.4. These results indicate that α₁- and α₂-adrenoceptor drugs' thermoregulatory actions are similar in event and suggest that both subtypes of α-adrenoceptor might be implicated functionally in a variety of thermoregulatory processes.

     

Estimation of the biophase concentration of noradrenaline at presynaptic α2-adrenoceptors in brain slices

Summary The aim of the present study was to determine the local concentrations of noradrenaline existing at presynaptic ₂-adrenoceptors during electrical pulse train stimulation of brain slices at different frequencies. The experiments are based on the assumption that the concentration of released noradrenaline at the ₂-adrenoceptors exerting a certain autoinhibition should be equal to the concentration of exogenous noradrenaline causing the same inhibition under conditions in which any influence of the released transmitter is excluded. In order to avoid autoinhibition, hippocampus and cortex slices of the rabbit and the rat, prelabelled with [³H]noradrenaline and superfused in presence of an uptake inhibitor, were electrically stimulated using 4 pulses delivered at 100 Hz (POP stimulation). Exogenous noradrenaline diminished the overflow of tritium elicited by POP stimulation in a concentration-dependent manner. In rabbit brain tissues the EC₅₀ value and maximum inhibition of noradrenaline release were found to be approximately 6 nmol/l and more than 95%, respectively, whereas in rat tissues the corresponding values were between 20 and 30 nmol/l and approximately 90%. When electrical stimulation was performed with trains of 36 pulses delivered at 0.1, 0.3 or 3 Hz in absence or presence of an uptake inhibitor, the ₂-adrenoceptor antagonist yohimbine (1 or 10 mol/l) enhanced the evoked tritium overflow in a manner which was dependent on the frequency of stimulation and on blockade of the re-uptake mechanism. The facilitatory effects of yohimbine reflected an extent of autoinhibition which was between 53% (36 pulses/0.1 Hz, no uptake inhibitor) and 85% (36 pulses/3 Hz, uptake inhibitor present) in rabbit and between 16% (36 pulses/0.3 Hz, no uptake inhibitor) and 71% (36 pulses/3 Hz, uptake inhibitor present) in rat brain slices. Accordingly, the corresponding estimated biophase concentrations of noradrenaline were generally higher in rat than in rabbit tissues (they were between 32.5 and 74.5 or 5.1 and 51.6 nmol/l in the presence or absence of an uptake inhibitor, respectively, in the rat, and between 15 and 23.1 or 6.1 and 18.6 nmol/l in the rabbit). The observed frequency dependence of the effect of re-uptake blockade on the calculated biophase concentrations of noradrenaline would be compatible with the idea of a dependence of the effectiveness of the re-uptake mechanism on the firing rate of the neurone in being more effective at lower frequencies. Moreover, the stikingly low biophase concentrations of noradrenaline suggest that also in brain tissue noradrenaline causes lateral inhibition of release as has recently been shown for guinea-pig vas deferens. Send offprint requests to C. Allgaier at the above address     

Typical and atypical neuroleptics are potent antagonists at α1-adrenoceptors of the dorsal lateral geniculate nucleus

Summary The present electrophysiological studies examined the actions of neuroleptics at central ₁adrenoceptors in the rat. In single-cell recording experiments, typical and atypical neuroleptics, when administered systemically or locally (iontophoresis and pressure ejection), were found to be potent antagonists of activating ₁adrenoceptor responses in the dorsal lateral geniculate nucleus (dLGN). Doses of neuroleptics effective as antagonists at ₁adrenoceptors had very weak effects as muscarinic receptors in the dLGN. Since doses of neuroleptics employed in the present study were within the clinical range, it appears likely that central ₁adrenoceptors would be blocked during neuroleptic therapy in humans.     

Involvement of μ-opioid receptors and α-adrenoceptors in the immunomodulatory effects of dihydroetorphine

The present study investigated the effects of acutely administered dihydroetorphine on mitogen-stimulated lymphocytes proliferation and lyrnphokine production in mice.These immune functions were significantly suppressed by dihydroetorphine at 24μg·kg~(-1) and 128μ·g-kg~(-1) in a dose-dependent fashion.This study further examined the involvement of μ-opioid receptors and     

The role of α1- and α2-adrenoceptors in the coronary vasoconstrictor responses to neuronally released and exogenous noradrenaline in the dog

Summary 1. Coronary vasoconstriction was examined in response to the neuronal release of noradrenaline produced by bilateral carotid occlusion and the infusion of tyramine (5 – 50 Erg/kg/min i. v.) in anaesthetized dogs which had been vagotomized and treated with the -adrenoceptor antagonist propranolol (1.0 mg/kg i. v.). These responses were compared to those produced by the infusion of noradrenaline (0.1 – 0.5 g/kg/min i. v.). 2. Similar increases in late diastolic coronary resistance were produced by bilateral carotid occlusion (0.70 ± 0.25 mmHg min/ml), and intravenous infusions of tyramine, 20 g/kg/min (0.70 ± 0.12 mm Hg min/ml) and noradrenaline, 0.5 gg/kg/min (0.59 ± 0.11 mm Hg min/ml). 3. Selective antagonism at ₁-adrenoceptors with prazosin (0.5 mg/kg i. v.) attenuated the coronary constrictor response to bilateral carotid occlusion (0.36 ± 0.09 mm Hg min/ml), tyramine (0.12 ± 0.06 mm Hg min/ml) and noradrenaline (0.18 ± 0.07 mm Hg min/ml). Antagonism at ₂-adrenoceptors with idazoxan (1 mg/kg i. v.) attenuated the coronary vasoconstriction produced by bilateral carotid occlusion (0.30 ± 0.06 mmHg min/ml), tyramine (0.17 ± 0.08 mmHg min/ml) and noradrenaline (0.12 ± 0.03 mm Hg min/ml). Combined antagonism at both ₁- and ₂-adrenoceptors with prazosin and idazoxan abolished the responses to bilateral carotid occlusion, tyramine and noradrenaline. 4. These results show that coronary vasoconstriction produced by either neuronally released or exogenous noradrenaline is mediated by both ₁ and ₁-adrenoceptors. It appears that in the coronary resistance vessels of the dog postjunctional ₁- and ₂-adrenoceptors are both innervated by sympathetic nerves.     

Involvement of α1- and α2-adrenoceptors in the responses of proximal and distal segments of the canine saphenous vein to exogenous and endogenous noradrenaline

Summary In the present study the relationship between adrenergic nerve terminals and postjunctional -adrenoceptors mediating the responses to the endogenous transmitter was compared at proximal and distal levels of the canine saphenous vein.Concentration-response curves to noradrenaline and to tyramine as well as frequency-response curves to electrical stimulation were compared at both levels of the vessel, in the absence and presence of either prazosin (100nmol·l^–1) or yohimbine (100nmol·l^–1) The influence of inhibition of neuronal uptake by cocaine (12 µmol·l^–1) on the responses to noradrenaline in the presence of prazosin (56 nmol·l^–1) or yohimbine (20 nmol·l^–1) was compared at the proximal level. The results show that, at the proximal level, the maximal responses to electrical stimulation and tyramine reached 80.1±2.2 (n = 18) and 74.2±1.9 (n = 18)%, respectively, of the maximal responses to noradrenaline, and 70.3±0.8 (n = 15) and 53.1 ± 1.2 (n = 14) %, respectively, at the distal level. Furthermore, the proximal strips were more sensitive to electrical stimulation than the distal ones. Prazosin had a much greater inhibitory effect on the contractile responses to noradrenaline than on those to electrical stimulation, at both levels. At proximal level, the shifts (to the right) of the concentration (frequency)-response curves (at EC₅₀) amounted to 0.58±0.02 (n = 16) and 0.18±0.02 (n = 8) log units, respectively (P<0.05), but, at the distal level, to 1.12±0.03 (n = 16) and 0.28±0.08 (n = 8) log units, respectively (P< 0.05). At the proximal level, yohimbine antagonizes about equally the responses to noradrenaline and the responses to electrical stimulation. However, at the distal level, the shift of the concentration-response curve to noradrenaline was much larger than that of the frequency-response curve to electrical stimulation [1.12±0.07 and 0.80±0.10 (n = 6) log units at EC₅₀, respectively (P<0.05)]. The leftward shift of the concentration-response curve to noradrenaline caused by cocaine was more pronounced in the presence of prazosin than in the presence of yohimbine: 0.95±0.15 and 0.69±0.12 (n = 12) log units, respectively (P<0.05).We conclude that, in the canine saphenous vein: 1) noradrenaline released from the adrenergic nerve terminals by electrical stimulation and by tyramine preferentially activates ₂-adrenoceptors at both proximal and distal levels; 2) the effectiveness of ₂-adrenoceptor stimulation is greater at the proximal than at the distal level; 3) ₁-adrenoceptors at the distal level seem to be different from those at the proximal one. Send offprint requests to S. Guimarães at the above address     

Presynaptic α-adrenoceptors and the action of tricyclic antidepressant drugs in behavioural despair in rats

The influence of the -adrenolytics, yohimbine and phentolamine, given in low doses thought to block presynaptic -adrenoceptors, on the action of the tricyclic antidepressants imipramine, nortriptyline and amitriptyline in the behavioural despair test was investigated in Wistar rats. Antidepressant drugs given in a single dose or for 2 weeks reduced immobility in rats. Yohimbine potentiated the effect of nortriptyline or amitriptyline administered in a single dose and the effect of imipramine and amitriptyline given for 2 weeks. The potentiating effect of phentolamine was observed only in rats receiving a single dose of nortriptyline. Yohimbine given chronically for 3 weeks together with antidepressants counteracted the effect of imipramine and nortriptyline on behavioural despair. Similarly clonidine abolished the reduction of immobility induced by antidepressants given in a single dose. It is concluded that the despair test is a behavioural model which is sensitive to the noradrenergic component of the drugs and that the blockade of presynaptic -adrenoceptors facilitates the action of tricyclic antidepressants in this test.     

Both α1- and β1-adrenoceptors in the bed nucleus of the stria terminalis are involved in the expression of conditioned contextual fear

BACKGROUND AND PURPOSE

The bed nucleus of the stria terminalis (BNST) is a limbic structure that is involved in the expression of conditioned contextual fear. Among the numerous neural inputs to the BNST, noradrenergic synaptic terminals are prominent and some evidence suggests an activation of this noradrenergic neurotransmission in the BNST during aversive situations. Here, we have investigated the involvement of the BNST noradrenergic system in the modulation of behavioural and autonomic responses induced by conditioned contextual fear in rats.

EXPERIMENTAL APPROACH

Male Wistar rats with cannulae bilaterally implanted into the BNST were submitted to a 10 min conditioning session (6 footshocks, 1.5 ma/ 3 s). Twenty-four hours later freezing and autonomic responses (mean arterial pressure, heart rate and cutaneous temperature) to the conditioning box were measured for 10 min. The adrenoceptor antagonists were administered 10 min before the re-exposure to the aversive context.

KEY RESULTS

L-propranolol, a non-selective β-adrenoceptor antagonist, and phentolamine, a non-selective α-adrenoceptor antagonist, reduced both freezing and autonomic responses induced by aversive context. Similar results were observed with {"type":"entrez-protein","attrs":{"text":"CGP20712","term_id":"874704353"}}CGP20712, a selective β₁-adrenoceptor antagonist, and WB4101, a selective α₁-antagonist, but not with ICI118,551, a selective β₂-adrenoceptor antagonist or RX821002, a selective α₂-antagonist.

CONCLUSIONS AND IMPLICATIONS

These findings support the idea that noradrenergic neurotransmission in the BNST via α₁- and β₁-adrenoceptors is involved in the expression of conditioned contextual fear.     

Modulation of motor activity by α1 and α2 stimulation in mice

Summary The influence of two -adrenoceptor agonists, clonidine and B-HT 920, on motor activity was tested in mice. Both, clonidine and B-HT 920 (2-amino-6-allyl-5,6,7,8-tetrahydro-4H-thiazolo-[4,5-d]-azepine) in the dose range 30–300 g/kg s.c. equieffectively inhibited exploratory activity. On the other hand only clonidine, which stimulates ₂- and ₂-adrenoceptors increased locomotor activity in mice treated with reserpine (5 mg/kg) and apomorphine (3 mg/kg) in the doses of 0.3 and 1 mg/kg i.p. The highly selective ₂-agonist B-HT 920 was ineffective under these conditions up to 30 mg/kg i.p. It is concluded, that in mice sedative -adrenoceptors are of the ₂- and excitatory of the ₁-type.     

Interactions of the enantiomers of 3-O-methyldobutamine with α- and β-adrenoceptors in vitro

Summary The enantiomers of 3-O-methyldobutamine, a metabolite of dobutamine, were evaluated for their - and -adrenoceptor mediated effects in vitro in a variety of isolated organs and in radioligand binding studies. Neither enantiomer of 3-O-methyldobutamine possessed ₁-adrenoceptor agonist activity in isolated guinea pig aorta. However, both enantiomers of 3-O-methyldobutamine were competitive ₁-adrenoceptor antagonists, with the (+)-enantiomer being approximately 10-fold more potent than the (-)-enantiomer as assessed either in guinea pig aorta or by displacement of ³H-prazosin binding from ₁-adrenoceptors in rat cerebral cortex. The ₁-adrenoceptor blocking activity of (+)-3-O-methyldobutamine was relatively potent and corresponded to a pA₂ of 7.33 in guinea pig aorta and a-log K _i of 7.72 in radioligand binding studies. Neither enantiomer of 3-O-methyldobutamine possessed ₂-adrenoceptor agonist activity in field-stimulated guinea pig ileum. Although (+)-3-O-methyldobutamine weakly inhibited the twitch response in field-stimulated guinea pig ileum, the response was not blocked by the selective ₂-adrenoceptor antagonist, yohimbine, and was found to result from weak anticholinergic activity (pA₂=5.06). Neither enantiomer of 3-O-methyldobutamine possessed ₁-adrenoceptor agonist activity in guinea pig atria, however the (+)-enantiomer was a weak noncompetitive antagonist at ₁-adrenoceptors. In contrast, both enantiomers of 3-O-methyldobutamine were weak ₂-adrenoceptor agonists in rat uterus, however these weak effects were not highly stereoselective, which was also confirmed in radioligand binding studies. The results of the present study indicate that 3-O-methyldobutamine is a potent and highly selective ₁-adrenoceptor antagonist, with minimal activity at ₂-, ₁- and ₂-adrenoceptors. It is hypothesized that the potent ₁-adrenoceptor antagonist activity of 3-O-methyldobutamine, which resides predominantly in the (+)-enantiomer, may play a role in the hemodynamic effects of dobutamine, by contributing, in part, to the decrease in total peripheral vascular resistance observed following administration of dobutamine.     

Effects of the Aconitum alkaloid mesaconitine in rat hippocampal slices and the involvement of α- and β-adrenoceptors

1. The effects of mesaconitine, the main alkaloid contained in Aconiti tuber, were investigated by use of extracellular recordings of stimulus-evoked population spikes and field excitatory postsynaptic potentials (e.p.s.ps) in the CA1 region of rat hippocampal slices.
2. At a concentration of 10 nM, mesaconitine evoked excitations, which were manifested as an increase in the amplitude of the orthodromic spike and the appearance of multiple spikes following the first postsynaptic spike, without affecting the magnitude of paired-pulse facilitation. The increase in spike amplitude was persistent and was not reversed by up to 90 min of washout. At concentrations of 30 and 100 nM, the alkaloid produced a biphasic effect, that is an excitation followed by an inhibition without having any effect upon the field e.p.s.p. At concentrations above 100 nM, mesaconitine suppressed the orthodromic population spike and the field e.p.s.p.
3. The excitatory effect was also observed when electrical stimulation was stopped completely during the application of mesaconitine (10 nM) and during the first 15 min of washout.
4. The enhancement of the population spike and the appearance of multiple spikes induced by mesaconitine (10–100 nM) were blocked by pretreatment with the β-adrenoceptor antagonists propranolol (1 μM) and timolol (1 μM), whereas the inhibitory effect was blocked by the α-adrenoceptor antagonists yohimbine (1 μM) and phentolamine (10 μM). However, when the β-adrenoceptor antagonist timolol was added 10 min after the application of mesaconitine, it failed to block the long-lasting enhancement of the spike amplitude and the appearance of multiple population spikes.
5. Application of the selective β-adrenoceptor agonist isoprenaline (500 nM) to the hippocampal slices induced an increase in the amplitude of the orthodromic population spike and elicited 2–3 additional spikes. Mesaconitine (10 nM) did not further potentiate this enhancement of the spike amplitude when added after a 15 min pretreatment with isoprenaline.
6. Perfusion of forskolin, which directly activates adenylate cyclase, enhanced the population spike. Mesaconitine had no additional effect when applied after pretreatment with forskolin.
7. It is concluded that the excitatory effects evoked by lower concentrations of the plant alkaloid mesaconitine are mediated by stimulation of β-adrenoceptors and the consequent activation of intracellular processes which lead to the long-lasting changes in excitability.

     

Phentolamine and hypoxia: modulation of contractility and α1-adrenoceptors in isolated rat atria

The hypoxia-induced effects on the binding sites and affinity constant of adrenoceptors, in the presence and absence of phentolamine, were determined for atrial membranes of hearts from normal and genetically hyperlipidaemic Yoshida (YOS) rats. Atrial function was also measured during normoxia and hypoxia, in the presence and absence of phentolamine.Hypoxia increased a1-adrenoceptor density in atrial membranes of normal rats (B_max 10.6 to 26.7 fmoles/mg protein). Phentolamine prevented the increase in the B_max of ₁-adrenoceptors and increased the equilibrium dissociation constant of these receptors (K _D 0.17 to 0.53 nmol/l). Beta-adrenoceptors did not change during hypoxia, but the B_max was slightly increased (26%) in the presence of phentolamine. Thus, the ₁/\ ratio increased from 0.40 in normoxia to 1.06 in hypoxia. In normoxic atria from YOS rats, the ₁/\ ratio was already elevated (0.86) in comparison to control rats (mainly due to a higher density of at-adrenoceptors in atrial membranes from YOS rats). This ratio was not modified by hypoxia (0.84), but decreased when phentolamine was present (0.30).Hypoxia reduced the force of contraction and increased diastolic tension of atria of normal rats, while the sinus rate was not significantly modified. Phentolamine abolished the increase in diastolic tension and reduced the negative effect of hypoxia on contractile force. In YOS rat atria, functional parameters were modified by hypoxia in a qualitatively similar way to that of normal rat atria.The observed increase in ₁-adrenoceptor density during hypoxia is in accordance with the results of experiments with animal models of the ischaemic heart and with findings in human heart failure. The possible therapeutic significance of these data is considered. Correspondence to: G. Fassina at the above address     

Role of α2-adrenoceptors in the regulation of intestinal water transport

1. The influence of the sympathetic nervous system on intestinal fluid transport by the jejunum and ileum of the anaesthetized rat was investigated under basal conditions and during active secretion induced by intra-arterial infusion of vasoactive intestinal peptide (VIP).
2. Intra-arterial infusion of noradrenaline (3, 10, 30 nmol min⁻¹, i.a.) and i.v. injection of the selective α₂-adrenoceptor agonist UK 14,304 (1 μmol kg⁻¹, i.v.) increased the rate of basal fluid absorption. The effect of UK 14,304 was blocked by yohimbine (10 μmol kg⁻¹, i.v). However, the selective α₁-adrenoceptor agonist phenylephrine (5 μmol kg⁻¹, i.v.) did not alter either the jejunal or ileal absorption rate.
3. The α₂-adrenoceptor antagonists yohimbine (0.3, 1.0, 3 and 10 μmol kg⁻¹, i.v.) and rauwolscine (10 μmol kg⁻¹, i.v.) decreased the basal absorption rate, while the α₁-adrenoceptor antagonist prazosin (3 μmol kg⁻¹, i.v.) was without effect. Intracerebroventricular injection of yohimbine (3 μmol kg⁻¹) caused a significant antiabsorptive effect in the jejunum but not ileum.
4. Peripheral chemical sympathectomy induced by pretreating animals with 6-hydroxydopamine (150 mg kg⁻¹, i.p., total dose) induced a trend towards impaired absorption in the jejunum and ileum.
5. The findings provide evidence that the sympathetic nervous system exerts tonic control on intestinal fluid transport and that the effect is mainly through peripheral α₂-adrenoceptors.
6. The subtype determination of α₂-adrenoceptors in modulating intestinal fluid transport was assessed by determining the effects of α₂-adrenoceptor agents on intestinal fluid secretion induced by i.a. infusion of VIP (0.8 μg min⁻¹).
7. Intravenous administration of UK 14,304 caused a dose-dependent reversal of the secretory phase of the VIP-induced response, but failed to restore fluid transport to the control level of net absorption. EC₅₀ values were 0.17 μmol kg⁻¹ in the jejunum and 0.22 μmol kg⁻¹ in the ileum.
8. The effect of UK 14,304 was blocked by the selective α_2A/D antagonist BRL 44408 and the non-selective α₂ antagonist yohimbine (each 10 μmol kg⁻¹). The selective α_2B/C antagonist ARC 239 (10 μmol kg⁻¹) did not affect the antisecretory action of UK 14,304. It is suggested that the α₂-adrenoceptors in the rat intestinal epithelium are the α_2D or α_2A-like subtype.

     

Heroin increases the density and sensitivity of platelet α2-adrenoceptors in human addicts

The density of platelet ₂-adrenoceptors, quantified by means of the binding of [³H]clonidine, and the aggregation response induced by adrenaline, was investigated in ten heroin addicts. The number of binding sites for [³H]clonidine was significantly increased during heroin abuse. Concomitantly, there was a potentiation of the adrenaline-induced platelet aggregation, which suggested that continuous heroin use (opiate dependence) is associated with supersensitive platelet ₂-adrenoceptors in human addicts. Spontaneous heroin withdrawal further increased in the same addicts the density of platelet ₂-adrenoceptors. These results suggest that platelet ₂-adrenoceptors may be used as a model to study receptor mechanisms of opiate dependence and withdrawal in humans