Vascular effects of ipsapirone are related with subtypes of alpha 1-adrenergic receptors

The aim of this study was to provide further evidence about the participation of Ó1-adrenoceptors in the vascular responses elicited by impsapirone. This 5-HT 1A agonist displayed vasodilator activity only when aortic rings were precontracted by Ó-adrenergic compounds. The relaxant effect was particulary evident when rings were precontracted with methoxamine (selective Ó1A-adrenergic agonist).On the other hand, ipsapirone but chloroethylclonidine (selective Ó1B-adrenergic antagonist), clearly displaced norepinephrine and methoxaminevasocontractile concentration-response curves to the right. Fanally, ipsapirone protected the Ó-adrenoceptors from the irreversible blockade provoked by phenoxybenzamine, as judged by thenorepinephrine contraction and stimulated phosphatidylinositil labeling. Accordingly the relaxant effect elicited by ipsapirone in aortic rings precontracted with Ó-adrenergic agonists and the protective action against blockade by phenoxybenzamine shown by this agent are proof of its ability to occupy Ó1-adrenoceptors. Specifically, Ó1A-adrenergic receptors seem to be involved in ipsapirone vascular effects, since this agent selectively relaxed aortic preparation precontracted with methoxamine and unlike chloroethylclonidine, clearly inhibited the contractile effect of this agonist. In summary, the present findings can be explained by accepting that ipsapirone may act as an antagonist at Ó1A-adrenoceptors

Central effects of an alpha 2-adrenergic antagonist on fetal lambs: a possible mechanism for hypoxic apnea.

In 12 chronically-prepared fetal lambs at 128-135 days gestation, a specific alpha 2-antagonist, L-657,743 (Merck) was infused into a lateral cerebral ventricle for periods of up to 24 h. Control infusions of artificial CSF had no effect. L-657,743 at a dose rate sufficient to block the actions of injected clonidine did not affect breathing incidence or episode duration, electrocortical activity, heart rate, arterial pressure or blood gases during normoxic conditions. However, 5 out of 6 preparations receiving L-657,743 while hypoxic continued to breathe during hypoxia instead of becoming apneic as normal. The central effects of alpha 2-adrenergic blockade appear to be specific to hypoxia. It is concluded that fetal hypoxic apnea may be mediated by inhibitory alpha 2-adrenergic receptors which can be blocked by the antagonist L-657,743.

Contribution of postsynaptic alpha 2-adrenoceptors to reflex sympathetic constriction of stenotic coronary vessels.

Increases in the activity of efferent cardiac sympathetic nerves by 35 +/- 9% were induced by 60 s bilateral occlusion of the common carotid arteries (BCO) in anesthetized dogs. Under control conditions the reflex rise in sympathetic nerve activity enhanced left ventricular pressure (115 +/- 4 mm Hg) by 47% and regional myocardial oxygen consumption (9.7 +/- 1.1 ml/min.100 g) by 56%. Simultaneously, end-diastolic circumflex coronary resistance (0.99 +/- 0.11 mm Hg.min.100 g/ml) decreased by 16%. After exhaustion of coronary dilator reserve by production of a severe coronary stenosis, BCO enhanced left ventricular pressure (107 +/- 4 mm Hg) by 49%, oxygen consumption of the poststenotic area (7.6 +/- 0.8 ml/min.100 g) increased by 21%, and circumflex coronary resistance (0.54 +/- 0.05 mm Hg.min.100 g/ml) also increased by 19%. The reflex increase in coronary resistance during BCO was abolished after infusion of the alpha 2-adrenoceptor antagonist rauwolscine (0.2 mg/kg i.v.). Administration of rauwolscine, however, did not prevent the reflex increase of left ventricular pressure and regional myocardial oxygen consumption. Comparable increases in poststenotic coronary resistance during BCO were found in dogs which either received propranolol (2 mg/kg i.v.) or in which the reflex rise in mean aortic pressure was limited to 13 +/- 3 mm Hg. In both experimental groups, rauwolscine also effectively prevented the BCO-induced rise in coronary resistance. In contrast, the reflex increase of total peripheral resistance was not significantly reduced by rauwolscine, but was blunted after additional administration of the selective alpha 1-adrenoceptor antagonist prazosin (1.2 mg/kg i.v.). We conclude that: 1) Poststenotic coronary vasoconstriction occurs during shortlasting increases in efferent cardiac sympathetic discharge within the physiological range. 2) This increase in poststenotic coronary resistance is significantly reduced after administration of the alpha 2-adrenoceptor antagonist rauwolscine. 3) In contrast to poststenotic coronary resistance, functionally innervated alpha 2-adrenoceptors are of minimal importance for the reflex increase in total peripheral resistance.

Lack of alpha-1-adrenergic receptor-mediated downregulation of angiotensin II receptors in neuronal cultures from spontaneously hypertensive rat brain.

Neuronal cells from Wistar Kyoto (WKY) and spontaneously hypertensive (SH) rat brains were established in culture to compare the expression of angiotensin II (Ang II) specific receptors and their regulation by norepinephrine (NE). Neurons from SH rat brains possess twice more Ang II specific receptors and expressed a proportional increase in Ang II stimulated [3H]-NE uptake compared with WKY neurons. NE caused a dose-dependent decrease in 125I-Ang II binding in WKY neurons, an effect not observed when neurons from SH rat brains were incubated with NE. These observations suggest that the lack of NE-induced downregulation of Ang II receptors in neuronal cultures is genetically regulated.

Evidence for an alteration in the microsomal Ca2+ release mechanism in senescent rat parotid acinar cells.

Previously, we have shown that Ca2+ mobilization following an alpha 1-adrenergic receptor stimulus is reduced in parotid acinar cells from senescent rats as a result of an altered ability of inositol 1,4,5-trisphosphate (IP3) to induce Ca2+ release from a non-mitochondrial, intracellular Ca2+ store (Ishikawa, Y., et al. Biochim. Biophys. Acta 968, 203-210). We have used this model to examine the IP3-induced Ca2+ release mechanism in these cells. 45Ca2+ efflux, after exposure to (-) epinephrine, from cells of young adult (3-6 months) rats was approx. 2-fold that observed from cells from older animals (approx. 24 months) either in the presence or absence of extracellular Ca2+. Similarly, cytosolic Ca2+ levels were greater in cells of young adult rats under these same incubation conditions. However, microsomal membrane preparations, from both age groups displayed similar IP3 binding sites (Kd approximately 90 nM, Bmax approximately 850 fmol/mg protein) and ATP-dependent Ca2+ transport ability (approx. 8 nmol/mg protein.min -1). These data suggest that there is an alteration in the IP3-induced Ca2+ release mechanism in microsomal membranes of parotid glands from senescent rats which may account for the decreased Ca2+ release seen after agonist stimulation of this tissue.

Regional variation of postjunctional alpha-adrenoceptor responses in the developing renal vascular bed of sheep.

Postjunctional alpha 1- and alpha 2-adrenoceptor vasoconstrictor responses were evaluated in isolated segments of main renal artery, segmental renal artery, and renal vein from fetal (130-138 d of gestation; term 145 d), newborn (3-15 d age), and nonpregnant adult sheep. Vascular rings were mounted at their optimal resting tension and responses to phenylephrine (alpha 1-adrenoceptor agonist) and guanabenz and UK14304 (both alpha 2-adrenoceptor agonists) were determined. Optimal resting tension increases with development in the main renal artery, segmental renal artery, and renal vein of sheep. Arterial vessels develop more isometric tension to alpha 1-adrenoceptor stimulation than to alpha 2-adrenoceptor stimulation, whereas venous segments develop similar isometric tension to alpha 1- and alpha 2-adrenoceptor stimulation. The segmental renal artery develops more isometric tension to alpha 2-adrenoceptor stimulation than the main renal artery. No large developmental differences exist among vessels in the sensitivity (concentration required for half maximal response, ED50) to alpha-adrenoceptor stimulation except for the renal vein with alpha 2-adrenoceptor stimulation. Maximum isometric tension corrected for vessel cross-sectional area decreases with age for all vessels with both alpha 1- and alpha 2-adrenoceptor stimulation. These findings may reflect developmental differences in receptor number and affinity or differences in vascular smooth muscle function. In addition, these data suggest that whereas both alpha 1- and alpha 2-adrenoceptors mediate vasoconstriction in the renal circulation, they may do so at different sites.

[Adrenergic coronary vasoconstriction during myocardial hypoperfusion]. / Vasoconstricção coronária adrenérgica nas situações de hipoperfusão miocárdica.

The intramural branches of the coronary arteries of dogs were studied using venilite casts. The intramyocardial vessels have a general pattern with straight branches that cross from the epicardium toward the endocardium. Many branches were observed throughout the extension of these vessels, mainly in the subendocardium, where there is a rich interconnecting plexus. Some branches divides immediately in the subepicardium. These anatomic findings concur with the observations that the subendocardium is more susceptible to ischemia, as it is dependent on an extensive vascularization. These anatomic findings indicate that during constant underperfusion, the alpha adrenergically mediated vasoconstriction of the subepicardial vessels have an unexpected beneficial effect because of decreased transmural steal.

Vasoconstricçäo coronária adrenérgica nas situaçöes de hipoperfusäo miocárdica / Adrenergic coronary vasoconstriction during myocardial hypoperfusion

Os autores estudam a distribuiçäo dos vasos coronarianos intramurais, através de moldes plásticos de vinilite, obtidos de coraçöes de cäo. Fporam estudados 6 moldes, de coraçöes injetados com vinilite e corroídos com ácido clorídrico. Os achados revelam que os vasos intramurais nascem de artérias situadas no epicárdio e que penetram em ângulo reto ou com pequena obliqüidade, na musculatura ventricular. Os vasos intramurais caminham até o subendocárdio, como um eixo, emitindo pequenos ramos em todo seu trajeto. Mais raramente observam-se ramos terminais para o subepicárdio. Ao nível do subendocárdio verifica-se uma extensa ramificaçäo vascular, que dá ao molde a aparência de uma penugem, com ampla anastomose para os vasos vizinhos, formando plexo vascular subendocárdico. Estes achados anatômicos estäo coerentes com a observaçäo de que a vascularizaçäo subendocárdica deva ser mais exuberante, para compensar o consumo de oxigênio nesta regiäo e a oposiçäo ao fluxo pela pressäo intracavitária. Com esta base anatômica é possível entender que em situaçöes de hipoperfusäo, o fluxo sangüíneo subendocárdico é mantido por vasoconstricçäo de vasos subepicárdicos. Há evidência na literatura de que esta açäo vasoconstrictora é mediada por receptores alfa adrenérgicos. A açäo de drogas vasodilatadoras que atuam nos pequenos vasos subepicárdicos, levaria a um desvio de fluxo com isquemia subendocárdica

Transient changes in cortical alpha 1 adrenoceptors and seizure threshold following electroconvulsive seizures in rats.

Alpha 1 adrenoceptor density (Bmax) is consistently decreased in actively spiking human cortical epileptic foci. Interpretation of these unique human data is limited because all surgical excisions are completed shortly after a period of active seizure discharge. To determine the temporal profile of seizure-induced changes in cortical alpha 1 adrenoceptors we examined rats primed by 15 daily electroconvulsive seizures (ECS). Since the noradrenergic system has an inhibitory effect on epileptic activity, we also measured the postictal rise in minimal ECS seizure threshold. Animals were sacrificed immediately before or at intervals after the last scheduled seizure. Cortical membranes were assayed using [3H]prazosin as specific radioligand. Repeated ECS produced an increase in the number of cortical alpha 1 sites from 4 to 24 h postictally, but following the last seizure there was a transient 'normalization' of alpha 1 receptor density which persisted for 3 h. The postictal ECS seizure threshold also remained elevated for a 2 h period. Both these transient postictal changes may in part result from activation of the central NA system. Decreased alpha 1 adrenoceptors in surgical specimens of spiking cerebral cortex may also be a secondary response to focal seizure activity.

Pharmacological characterization of alpha-adrenoceptors involved in nictitating membrane and pupillary responses to sympathetic nerve stimulation in cats.

Electrical stimulation of the preganglionic cervical sympathetic nerve produced frequency-related contraction of the nictitating membrane (NM) and dilation of the pupil in anesthetized cats. Observations of the effects of alpha-adrenoceptor blockade on these effectors were made simultaneously from the same preparations. All of the alpha 1-adrenoceptor antagonists tested produced a dose-related blockade of the NM with the relative potencies being prazosin greater than WB-4101 greater than phentolamine greater than phenoxybenzamine. In contrast, the iris dilator response was blocked by WB-4101 and phenoxybenzamine but was almost totally refractory to antagonism by doses of prazosin and phentolamine that reduced the evoked NM responses by more than 75% in these same preparations. Neither alpha 2-adrenoceptor (yohimbine or rauwolscine) nor beta-adrenoceptor (propranolol) antagonism produced significant inhibition of the activation of either organ. These results suggest that: 1) neural activation of the nictitating membrane is solely due to stimulation of alpha 1-adrenoceptors; 2) neither beta- nor alpha 2-adrenoceptors contribute significantly to nerve activation of either the nictitating membrane or iris dilator muscle in vivo; 3) the alpha-adrenoceptors on the dilator muscle that are activated neurally can not be classified pharmacologically as either alpha 1 or alpha 2 receptors.

[Effect of guanabenz on the peristaltic reflex and on the spontaneous rhythmic movements of the ileum isolated from the rabbit: do the alpha-2-adrenoreceptors constitute a homogenous population?]. / L'effet du guanabenz sur le réflexe péristaltique et sur les mouvements rythmiques spontanés de l'iléon isolé de Lapin: les alpha-2 adrénocepteurs constituent-ils une population homogène?

The inhibitory effect of the predominantly alpha-2 adrenoceptor agonist, guanabenz, on the peristaltic reflex and on the pendular movements of the rabbit isolated ileum was investigated. Guanabenz depressed or abolished the peristaltic reflex as well as the pendular movements. These effects were concentration-dependent. Guanabenz is much more potent inhibiting the peristaltic reflex (IC50 1 X 10(-7) M) than the pendular movements (IC50 1 X 10(-5) M). The choline ester, acetylcholine restored the peristaltic reflex and the anticholinesterase, eserine, restored the pendular movements previously abolished by guanabenz. During the blockade of the peristaltic reflex produced by guanabenz, the pendular movements were virtually not changed. It is therefore reasonable to suppose that the inhibitory effect of guanabenz reflects the different properties of alpha-2 adrenoceptors associated with cholinergic nerve terminals within the myenteric plexus and the longitudinal smooth muscle subserving the peristaltic reflex and the pendular movements.

Effects of stimulation of the ocular sympathetic nerves on IOP and aqueous humor flow.

Ocular sympathetic nerves were stimulated chronically in awake rabbits using electrodes unilaterally implanted on the cervical sympathetic trunk. IOP was measured by pneumatonometry and aqueous inflow was measured by fluorophotometry. In each animal, continuous trains of 1 msec pulses were delivered by means of a portable electrical stimulator. Experiments were spaced by 1 week recovery periods. Stimulation was varied over a range of amplitudes (5-15 V) and frequencies (3-12 Hz). Continuous sympathetic stimulation produced an immediate sharp decrease in IOP followed by a gradual rise to pre-stimulation values which were attained 60-90 min after onset. A rebound increase in IOP occurred when stimulation was terminated. The magnitude of the initial IOP drop, the delay in the return to pre-stimulation IOP, and the rebound rise in IOP subsequent to termination of electrical stimulation were proportional to the stimulation frequency. Maximal effects were observed at 12 Hz, and stimulation with 8-10 Hz for 180 min caused a sustained reduction in anterior chamber aqueous humor flow. Topical 2% phentolamine 1 hr before stimulation markedly reduced IOP and abolished the acute IOP changes observed in untreated stimulated animals. Topical 1% timolol did not affect either the initial IOP drop or the rebound; however, the IOP recovered during stimulation to values greater than pre-stimulation IOP. We conclude that in rabbits the beta-adrenergic effect of prolonged sympathetic nerve stimulation is to decrease aqueous flow. Chronic electrical stimulation in awake animals provides an experimental model for studying the role of the ocular sympathetic nerves.

Increase of alpha 2-presynaptic adrenoceptor response after neuronal uptake inhibition in vivo.

alpha 2-adrenoceptor antagonists, yohimbine or idazoxan (1 mg kg-1 i.p.), administered alone, did not change noradrenaline content in the central and peripheral tissues of the rat (hypothalamus, brain stem, frontal cortex and heart). The inhibition of neuronal uptake by desipramine (DMI) administered alone or prior to alpha 2-adrenoceptor antagonists did not affect the neurotransmitter content either. alpha-methyl-p-tyrosine (alpha-MT) 6 hr before sacrifice, induced a marked disappearance of the noradrenaline content, greater in central nervous tissues than in heart. When the catecholamine synthesis was inhibited by alpha-MT, neither alpha 2-adrenoceptor antagonists nor DMI at the dose used, significantly changed the disappearance rate of noradrenaline in any of the tissues studied. Under these experimental conditions, however, the combination of DMI plus alpha 2-adrenoceptor antagonists significantly decreased the neurotransmitter content in all tissues when the values were compared with the control or DMI-treated groups. The present results might suggest evidence in favour of a functional coupling between presynaptic alpha 2-autoreceptors and noradrenaline uptake mechanism.

Positive chronotropic responses produced by alpha-adrenoreceptors in the pithed rat.

1 Phenylephrine (1-100 microgram/kg, intravenously) produced dose-dependent increases in heart rate and blood pressure in the pithed rat. 2 The positive chronotropic response to phenylephrine (10 microgram/kg) was reduced in a dose-dependent manner by propranolol (0.01-0.3 mg/kg), but higher doses of propranolol (up to 3 mg/kg) did not reduce the response by more than about 50%. The residual response was virtually abolished by phentolamine (0.3 mg/kg) or prazosin (3 microgram/kg). Labetalol (3 mg/kg) which has both alpha- and beta-blocking activity, also abolished the positive chronotropic response. 3 The pressor response to phenylephrine (1-30 microgram/kg) was enhanced by propranolol (1 mg/kg) and abolished by phentolamine (1 mg/kg) and prazosin (30 microgram/kg). Labetalol (3 mg/kg) reduced the response to phenylephrine by 73%. 4 Propranolol (0.3 mg/kg) completely blocked the chronotropic and vasodepressor effects of isoprenaline (0.1 microgram/kg). 5 It is concluded that phenylephrine acts on both alpha 1- and beta 1-adrenoreceptors to produce an increase in heart rate, on alpha 1-adrenoreceptors to produce vasoconstriction and on beta 2-adrenoreceptors to produce vasodilation. This latter effect is usually masked by the predominant vasoconstrictor action.

Alpha-adrenergic vasoconstriction in arterial and arteriolar sections of the canine coronary circulation.

Using a method which allowed the in-situ measurement of segmental coronary vascular resistances, the reaction of arterial and arteriolar sections of the coronary vascular system to alpha-receptor stimulation was studied i anaesthetized dogs. The left coronary artery was cannulated, and the perfusion pressure was kept constant. Allowance was made for extravascular and metabolic influences of alpha-stimulation on coronary vascular resistances. On an average, submaximal alpha-stimulation with xylometazoline increased the arterial resistance by about 60% and the arteriolar resistance by about 90%. The cannulation of the left coronary artery increased the sympathetic reactivity of the arterioles. Moreover, xylometazoline increased the extravascular component of the coronary vascular resistance by about 4%. Thus it can be assumed that under normal in-vivo conditions alpha-receptor vasoconstriction might be less different in coronary arteries and arterioles. Since the arterial resistance ranges from 20% to 50% of total coronary resistance, a sympathetic vasoconstriction of this vascular section might lead even to a critical limitation of coronary blood flow. On the other hand, a predominant constriction in arterioles leads to an increase in peripheral coronary pressure, i.e. to a "reverse coronary steal phenomenon".

Alpha-adrenoceptor-mediated modulation of 5-hydroxytryptamine release from rat brain cortex slices.

Slices of rat brain cortex preincubated with 3H-5-hydroxytryptamine were superfused with physiological salt solution and stimulated electrically, or they were superfused with Ca2+-free solution containing 25 mM K+ and stimulated by introduction of 1.3 mM CaCl2 for 2 min. 1. The electrically evoked 3H overflow was decreased by noradrenaline and increased by phentolamine in a concentration-dependent manner. 2. Phentolamine caused a parallel shift to the right of the concentration-response curve of noradrenaline for its inhibitory effect on impulse-evoked 3H overflow; by contrast, it left the inhibitory effect of unlabelled 5-hydroxytryptamine on the electrically evoked 3H overflow unaffected. 3. Propranolol did not alter the inhibitory effect of noradrenaline on impulse-induced 3H overflow. 4. The increasing effect of phentolamine on the electrically evoked 3H overflow was not modified by paroxetine. 5. Cocaine (at a concentration which almost completely blocks the uptake of 3H-noradrenaline but only partially that of 3H-5-hydroxytryptamine into cortex slices) decreased impulse-evoked 3H overflow. This effect was abolished by phentolamine. 6. In the presence of tetrodotoxin throughout superfusion, the Ca2+-evoked 3H-overflow from slices superfused with Ca2+-free solution was inhibited by noradrenaline and increased by phentolamine. These findings suggest that the terminal serotoninergic fibers of rat brain cortex possess alpha-adrenoceptors, activation of which by exogenous or endogenous noradrenaline leads to inhibition of the release of 5-hydroxytryptamine.

In vitro characterization of adrenergic receptors mediating extrusion of preformed sebum from preputial gland of rat.

Extrusion of preformed sebum from the preputial glands of rat after phenylephrine and adrenaline treatment, and its inhibition by alpha-receptor blocking agents under in vitro conditions, shows that secretory response of the glands is influenced by alpha-adrenergic receptors, and isoproterenol--a beta-agonist--is not effective in elicitation of exudation of secretion from the preputial gland.

Structure activity relationships between catecholamines and the alpha-adrenergic receptor responsible for the aggregation of human platelets by epinephrine.

PEA is a potent inhibitor (Ki approximately 13 microM) of human platelet aggregation induced by epinephrine. This led us to perform an SAR study of a congeneric series of compounds in an effort to identify the molecular components of epinephrine critical to its aggregating effect upon human platelets. Phenylethanolamine was similar to PEA in inhibitory potency. However, hydroxylation of the phenyl ring diminished the inhibitory effect (Ki tyramine approximately 87 microM; Ki octopamine approximately 88 microM). Dopamine, the weakest inhibitor (Ki approximately 150 microM), was a partial agonist capable of inducing platelet aggregation in some samples of PRP. The order of potency of catecholamines as aggregating agents was epinephrine greater than norepinephrine greater than Epinine greater than dopamine. Phenylephrine, the prototype alpha-agonist, did not induce aggregation but was a potent inhibitor (Ki approximately 12 microM) of the aggregation induced by epinephrine. Isoproterenol, the prototype beta-agonist, was neither an aggregant nor an inhibitor of epinephrine-induced platelet aggregation. These findings suggest that the binding of epinephrine to the alpha-adrenergic receptor responsible for platelet aggregation is accomplished by the N-methyl amino group whereas intrinsic aggregating activity is a function of the catechol moiety.