Two distinct serotonin receptors co-mediate non-photic signals to the circadian clock.

Several lines of evidence indicate that serotonin type 7 (5-HT7) receptors play a critical role for non-photic resetting of the mammalian circadian clock; however, the contributions of other types of 5-HT receptors to non-photic entrainment are not yet clarified. The present study demonstrates that MKC-242, a selective 5-HT1A receptor agonist, can evoke a non-photic-like phase-response in hamsters in vivo. This phase-shifting response to MKC-242 was antagonized not only by the selective 5-HT1A receptor blocker WAY100635 but also by the selective 5-HT7 receptor blocker DR4004. These suggest that synchronous activation of 5-HT1A and 5-HT7 receptors mediates non-photic signals to the hamster circadian clock.

Pharmacological characterizations of adrenergic receptors in human adipocytes.

Three types of adrenergic receptors, beta, alpha-1, and alpha-2, were identified in human adipocytes, isolated from properitoneal adipose tissue, using both the binding of radioactive ligands and the effects of adrenergic agents on receptor-specific biochemical responses. Adrenergic binding studies showed the following results: [(3)H]dihydroalprenolol binding (beta adrenergic) B(max) 280 fmol/mg protein, K(D) 0.38 nM; [(3)H]para-aminoclonidine binding (alpha-2 adrenergic) B(max) 166 fmol/mg protein, K(D) 0.49 nM; [(3)H]WB 4101 binding (alpha-1 adrenergic) B(max) 303 fmol/mg protein, K(D) 0.86 nM. In adipocytes from subcutaneous adipose tissue, [(3)H]dihydroergocryptine binding indicated the presence of alpha-2 but not alpha-1 receptors. Beta and alpha-2 adrenergic receptors appeared to be positively and negatively coupled to adenylate cyclase, respectively. Cells or cell membranes were incubated with epinephrine (10 muM) alone and in combination with the antagonists yohimbine (alpha-2) and prazosin (alpha-1). Epinephrine alone prompted a modest increase in adenylate cyclase activity, cyclic AMP, and glycerol release, an index of lipolysis. Yohimbine (0.1 muM) greatly enhanced these actions whereas prazosin was without effect. The beta agonist, isoproterenol, stimulated glycerol release, whereas the alpha-2 agonist, clonidine, inhibited lipolysis and cyclic AMP accumulation. To assess further alpha-1 receptors, cells were incubated with [(32)P]phosphate and epinephrine (10 muM) alone and in combination with prazosin and yohimbine. Epinephrine alone caused a three- to fourfold increase in (32)P incorporation into phosphatidylinositol. Prazosin (0.1 muM) blocked this action whereas yohimbine (0.1 muM) was without effect. Thus, in a homogeneous cell preparation, the human adipocyte appears to have three different adrenergic receptors, each of which is coupled to a distinct biochemical response.

Clazosentan (Actelion).

Clazosentan, an endothelin ETA antagonist, is under development by Actelion (formerly Axovan), under license from F Hoffman-La Roche, for the potential prevention of cerebral infarction and ischemia induced by cerebral vasospasm following subarachnoid hemorrhage. Results from the phase IIb portion of a phase IIb/III clinical study are expected in the first half of 2006.

QSAR study for a novel series of ortho disubstituted phenoxy analogues of alpha1-adrenoceptor antagonist WB4101.

On the basis of the affinities at the alpha1a-, alpha1b- and alpha1d-adrenoceptors and the 5-HT1A receptor of a previous series of sixteen 2-[(2-phenoxyethyl)aminomethyl]-1,4-benzodioxanes ortho monosubstituted at the phenoxy moiety, a number of ortho disubstituted analogues were designed, synthesized in both the enantiomeric forms and tested in binding assays on the same receptors. The affinity values of the new compounds 1-11 were compared with those of the enantiomers of the 2,6-dimethoxyphenoxy analogue, the well-known alpha1 antagonist WB4101, and of the ortho monosubstituted derivatives, suggesting some distinctive aspects of the interaction of the phenoxy moiety, in particular with the alpha1a-AR and the 5-HT1A receptor, of the monosubstituted and the disubstituted compounds. A classical quantitative structure-activity relationship (Hansch) analysis was applied to the whole set of the S enantiomers of the ortho mono- and disubstituted WB4101 analogues (26 compounds), finding a very good correlation for the alpha1a affinity. For this latter, a significant parabolic relationship was also found with the volume of the two ortho substituents. Diametrically opposite, the same relationships for the 5-HT1A exhibit low or insignificant correlation coefficients.

Recombinant human alpha 2-adrenoceptor subtypes: comparison of [3H]rauwolscine, [3H]atipamezole and [3H]RX821002 as radioligands.

Kinetic, saturation and competition binding assays were employed to optimize and validate radioligand binding methods for characterization of recombinant human alpha 2-adrenoceptor subtypes and for screening of new subtype-selective ligands. Stable transfected lines of Shionogi 115 mouse mammary tumour cells (S115) and three structurally different antagonist radioligands, [3H]rauwolscine, [3H]atipamezole and [3H]RX821002, were used. Specificity of alpha 2-adrenergic binding was defined with 100 microM (-)-adrenaline. Steady-state was reached with all three radioligands within 15-30 min at 25 degrees C, and the binding was rapidly reversible. The receptor affinities (alpha 2-C10) were highest in glycylglycine, almost equally high in K(+)-phosphate, and lowest in Tris buffer for all three [3H]-ligands. This was mainly caused by different association rates. [3H]RX821002 was bound with high affinity and similar kinetic properties to all three alpha 2-adrenoceptor subtypes in K(+)-phosphate buffer, and had the highest proportion of specific binding (96-98%). [3H]RX821002 and K(+)-phosphate buffer were subsequently used in competition assays. The rank order of affinity of compounds selective for alpha 2-adrenoceptor subtypes was alpha 2-C10 > alpha 2-C4 > alpha 2-C2 for oxymetazoline, alpha 2-C4 > alpha 2-C2 > alpha 2-C10 for prazosin and alpha 2-C2 > alpha 2-C4 > alpha 2-C10 for chlorpromazine. The drug affinities (Ki values) determined in this system were in close agreement with earlier results with [3H]rauwolscine in Tris buffer (r = 0.94). Agonist competition for [3H]RX821002 binding was biphasic in K(+)-phosphate buffer supplemented with 10 mM MgCl2, indicating functional coupling of receptors to G-proteins. Accordingly high-affinity binding of the agonists (-)-noradrenaline and UK14,304 was eliminated by 10 microM Gpp(NH)p in the assays. Our results confirm that [3H]RX821002 is a suitable radioligand for the characterization of all three human alpha 2-adrenoceptor subtypes and for the determination of the subtype-selectivity of new alpha 2-adrenoceptor agonists and antagonists.

Non-adrenergic binding sites for the "alpha 2-antagonist" [3H]idazoxan in the rabbit urethral smooth muscle. Pharmacological and biochemical characterization.

In the present study, pharmacological and biochemical binding characteristics of [3H]idazoxan, an originally thought alpha 2-adrenoceptor antagonist, have been determined in smooth muscle of rabbit urethra. It is shown that [3H]idazoxan labels with high affinity non-adrenergic binding sites. Specific binding of [3H]idazoxan is inhibited by compounds possessing an imidazoline or a guanidinium moiety whereas phenylethanolamines and classical alpha 2-antagonists are ineffective competitors which suggests an imidazoline-preferring binding site. However, imidazolidines such as clonidine and paminoclonidine are poorly effective, which differs considerably from pharmacological characteristics of imidazoline binding sites previously reported in the central nervous system. In addition, it is shown that K+ and Mn2+ inhibit [3H]idazoxan binding in a competitive and non-competitive manner, respectively. Other cations such as Na+, Li+ and Mg2+ have no significant effect. It is shown that K+ accelerates the dissociation of [3H]idazoxan binding while Mn2+ does not produce any modification. These results suggest that K+ may bind to an allosteric site, while Mn2+ may bind with a membrane component susceptible to alter [3H]idazoxan binding sites.

Different affinities of alpha 2-agonists for imidazoline and alpha 2-adrenergic receptors.

It has recently been shown that imidazoline alpha 2-adrenergic agonists, such as clonidine and UK 14,304, selectively bind to both alpha 2- and imidazoline receptors in basolateral membranes from rabbit renal proximal tubule. In order to define the relative affinity of three antihypertensive alpha 2-agonists for the two classes of receptors, we performed competition studies of imidazoline alpha 2-antagonist 3H-RX 781094 and nonimidazoline antagonist 3H-rauwolscine binding to basolateral membranes from rabbit proximal tubule. The order of potency for inhibition of radioligand binding to basolateral membranes was rilmenidine greater than clonidine greater than guanfacine and clonidine greater than guanfacine greater than rilmenidine for 3H-RX 781094 and 3H-rauwolscine binding, respectively. These data show that not only clonidine, but also rilmenidine and guanfacine, drugs usually used as specific alpha 2-agonists, bind to both alpha 2- and imidazoline receptors. The higher affinity of these molecules for one or the other class of receptors could explain their different capacity to induce hypotension and side effects.

Hypothalamic alpha 2A-adrenoceptors stimulate growth hormone release in the rat.

Oxymetazoline, the relatively selective alpha 2A-adrenoceptor agonist (with more than 60-fold selectivity over the alpha 2B-adrenoceptor subtype), was administered into the lateral ventricle (i.c.v.) of rats and plasma growth hormone (GH) levels were measured. Oxymetazoline was more potent to release GH after i.c.v. administration than was clonidine; 0.01 microgram i.c.v. oxymetazoline already caused a significant release of GH, while at least 0.1 microgram clonidine had to be administered to cause a similar response. The dose-response curve was of an inverted U shape since with 10 micrograms of oxymetazoline the plasma GH did not rise. When oxymetazoline was injected i.c.v. to rats with somatostatin fibres to the median eminence transected by an anterolateral cut in the hypothalamus there was a significant rise in plasma GH, suggesting that oxymetazoline stimulated GHRH rather than inhibited somatostatin release. Pretreatment with CH-38083 (7,8-(methylenedioxy)-14-alpha-hydroxy-alloberban HCl, selective for alpha 2-adrenoceptors but not differentiating between alpha 2A and alpha 2B subtypes), prevented the plasma GH rise normally elicited by 1 microgram i.c.v. oxymetazoline. The alpha 2A- and alpha 1-selective adrenoceptor antagonist, WB-4101 (2-(2,6-dimethoxyphenoxyethyl)-aminomethyl-1,4-benzodioxane hydrochloride), prevented the GH rise normally induced by oxymetazoline while prazosin, the alpha 2B- and alpha 1-selective adrenoceptor antagonist, prolonged the elevation occurring in the control rats between 30 and 60 min after oxymetazoline injection. Since both prazosin and WB-4101 are alpha 1-adrenoceptor antagonists but differ in their action on alpha 2A and alpha 2B subtypes as well as in their action on oxymetazoline-induced GH secretion, the antagonist studies suggest that oxymetazoline stimulates GH release through activation of alpha 2A-adrenoceptors stimulatory to GHRH release, and not by an action through alpha 2B- or alpha 2C- or alpha 1-adrenoceptors. Since WB-4101 also antagonized clonidine action on GH release we also suggest that the major component may be the stimulation of the alpha 2A-adrenoceptors in the clonidine action on GH release.

Centrally induced impairment of the hypotensive effects of R 28935 and R 29814 by prazosin in anaesthetized cats.

Prazosin (3 micrograms/kg) infused prior to erythro-1-(1-[2-(1,4-benzodioxan-2-yl)-2-hydroxyethyl]-4-piperidyl)-2-benzimidazolinone (R 28935) into the left vertebral artery (v.a.) of chloralose-anaesthetized cats diminished the hypotensive response to R 28935 (3 micrograms/kg, v.a.). A similar i.v. treatment did not alter the central hypotensive effect of R 28935 (micrograms/kg, v.a.). After prazosin (3 micrograms/kg, v.a.) there was a parallel shift to the right of the dose-response curve of R 28935 (v.a.). The central hypotensive effect of the threo isomer (R 29814; 30 micrograms/kg) was also diminished by the previous administration of prazosin (3 micrograms/kg, v.a.). These results indicate the involvement of central alpha 1-adrenoceptors in the mechanism of action of R 28935 and R 29814.

Prazosin modulates the firing pattern of dopamine neurons in rat ventral tegmental area.

Previous studies have indicated a noradrenergic modulation of midbrain dopamine cell activity. The effects of systemic administration of the alpha 1-adrenoceptor antagonist prazosin and the alpha 2-antagonist idazoxan on midbrain dopamine cell firing were now studied with extracellular recording from single dopamine neurons in the ventral tegmental area of chloral hydrate-anaesthetized male rats. Prazosin (0.15-0.6 mg/kg i.v.) dose dependently decreased burst firing and regularized the firing pattern of dopamine neurons, while the firing rate was unaffected. The prazosin-induced effects were abolished by pretreatment with reserpine. Idazoxan (0.5-2.0 mg/kg i.v.) increased firing rate and burst firing and made the firing pattern less regular, probably by increasing adrenergic transmission via blockade of presynaptic alpha 2-adrenoceptors. The effects of idazoxan were blocked by prazosin. The present results indicate that noradrenergic neurons modulate the dopamine cell firing pattern via excitatory postsynaptic alpha 1-adrenoceptors. This mechanism might be involved in the pathogenesis and pharmacological treatment of schizophrenia.

The positive inotropic effect of alpha 1A-adrenoceptor stimulation is inhibited by 4-aminopyridine.

This study was designed to determine if 4-aminopyridine, a reported inhibitor of the transient outward K+ current (Ito), alters the inotropic actions elicited via stimulation of WB4101- or chloroethylclonidine-sensitive receptors in rat myocardium. WB4101 (N-[2-(2, 6-dimethoxyphenoxy)ethyl]-2,3-dihydro-1,4-benzodioxin-2-m ethanamine) is a competitive antagonist that is selective for alpha 1A- and alpha 1C-adrenoceptors, while chloroethylclonidine is an irreversible blocker that is reported to antagonize alpha 1B-, alpha 1C-, and alpha 1D-adrenoceptor binding. Inotropic effects of the alpha 1-adrenoceptor agonist phenylephrine were examined in isolated left atrial and papillary muscle before and after addition of 4-aminopyridine, and before and after addition of 4-aminopyridine in preparations pretreated with chloroethylclonidine or WB4101. In addition, effects of phenylephrine were examined before and after treatment with staurosporine (an inhibitor of protein kinase C) in chloroethylclonidine-pretreated preparations. Phenylephrine (10 microM) elicited a sustained positive inotropic response in left atria and a triphasic inotropic action in papillary muscle (transient positive and negative inotropic components preceding a sustained positive inotropic response). 4-Aminopyridine (1.0, 1.7, 3.0 mM) reduced the sustained positive inotropic responses in the absence of antagonists and in chloroethylclonidine-pretreated preparations. However, in the presence of 10 nM WB4101, 4-aminopyridine had no effect on the remaining inotropic actions of phenylephrine. The sustained positive inotropic response to the alpha 1-agonist in chloroethylclonidine-pretreated preparations was not inhibited by 100 nM staurosporine. These data suggest that the sustained positive inotropic actions of alpha 1A-adrenoceptor stimulation in rat atrial and ventricular myocardium are mediated via non-protein kinase C-associated reductions in Ito.

Pertussis toxin blocks the effects of alpha 2-agonists and antagonists on locus coeruleus activity in vivo.

This study assessed the effects of pertussis toxin, which is known to inactivate G proteins and therefore to block receptors linked to G proteins, on electrophysiological activity of the locus coeruleus in vivo. Pertussis toxin was injected into the lateral cerebral ventricle of rats, and locus coeruleus activity was then recorded. Compared to vehicle-injected control animals, pretreatment with pertussis toxin markedly increased the spontaneous firing rate of locus coeruleus neurons. In addition, the alpha 2-antagonist idazoxan was no longer able to augment either spontaneous or evoked locus coeruleus activity after pretreatment with pertussis toxin. Finally, pretreatment with pertussis toxin made locus coeruleus neurons resistant to inhibition by the alpha 2-agonist clonidine. These results are consistent with the view that pertussis toxin blocks alpha 2-receptors, receptors linked to G proteins, in vivo.

Anxiogenic-like effects of yohimbine and idazoxan in two behavioral situations in mice.

In a light-dark choice situation, the alpha-2 adrenoceptor antagonist idazoxan shows anxiogenic-like effects, which cannot be blocked by the alpha-2 adrenoceptor agonist clonidine, or by the benzodiazepine receptor antagonist Ro 15-1788. In a conditioned conflict situation, both idazoxan and the alpha-2-adrenoceptor yohimbine show anxiogenic-like effects; the effect of idazoxan could not be blocked by clonidine or Ro 15-1788. These data suggest that systems other than alpha-2 adrenoceptors or benzodiazepine receptors must be found to explain these anxiogenic-like properties.

Rabbit cerebral cortex 5HT1a receptors.

Selective 5HT1a agonist binding to membranes from rabbit cerebral cortex was concentration-dependent and saturable; the Kd was 1.1 nM and Bmax of 480 fmols/mg protein. Scatchard as well as Hill plots were linear; the Hill coefficient was 0.96, suggesting a single, non-interacting binding site. Agonist binding was inhibited in a concentration-dependent fashion by gamma S GTP, a result consistent with the coupling of this binding site to the G protein signal transduction system. In competition experiments involving agonist and a series of agents with known affinities and specificities at 5HT1a receptors, a rank order relationship was found consistent with this binding site being a 5HT1a binding site. Direct comparisons of agonist and antagonist binding at rat cerebral cortex 5HT1a receptors and cloned human 5HT1a receptors also suggested that the rabbit binding site belongs to the 5HT1a class. The only rank order anomalies were with methiothepin in rabbit cerebral cortex, where a comparatively high Ki was observed and with buspirone in cloned human 5HT1a receptor, where a low Ki was determined; these anomalies bear further study in light of the comparative pharmacology of 5HT1a receptors. Finally, the natural product parthenolide was tested for affinity in the rabbit, rat, and human systems, where it uniformly was unable to displace agonist, suggesting that the 5HT1a receptor is not a target for this compound. Overall, these results suggest that a functional 5HT1a receptor exists in rabbit cerebral cortex.

[Selectivity of alpha 2-adrenergic agonists for the imidazoline-guanidine and alpha 2-adrenergic receptors]. / Sélectivité des agonistes alpha 2-adrénergiques pour les récepteurs imidazolinique-guanidiniques et alpha 2-adrénergiques.

Imidazolines have been proposed as highly selective drugs for alpha 2-adrenergic receptors. However, we have recently showed that the imidazoline ligand 3H-RX 781094 (idazoxan) binds to both alpha 2-receptors and imidazoline guanidinium receptive substance (IGRS) in rabbit renal proximal tubule. Binding of 3H-RX 781094 to the purified basolateral membranes (15-fold enriched in Na-KATPase activity) was rapid (t 1/2 = 5 mn.) reversible (t 1/2) = 4 mn.), saturable and of high affinity. Scatchard analysis of equilibrium binding data showed that 3H-RX 781094 labels 566 +/- 118 fmol/mg of proteins of binding sites with an apparent dissociation constant (Kd) of 1.45 +/- 0.14 nM. On the other hand, the non imidazoline ligand 3H-rauwolscine binds only to the alpha 2-adrenergic receptors with a maximal density of 155 +/- 28 fmol/mg of protein and a Kd of 11.5 +/- 1.5 nM. In order to define the relative affinity of the alpha-2-agonists, clonidine, rilmenidine and guanfacine for the two classes of receptors, we performed competition studies of the alpha 2-antagonists 3H-RX 781094 (imidazoline) and 3H-rauwolscine (non imidazoline) binding to basolateral membranes from rabbit proximal tubule. The order of potency for inhibition of the two radioligand binding was rilmenidine greater than clonidine greater than guanfacine for 3H-RX 781094 and clonidine greater than guanfacine greater than rilmenidine for 3H-rauwolscine. Therefore, rilmenidine displayed a higher affinity for IGRS than for alpha 2 adrenergic receptors; on the other hand, clonidine and guanfacine preferentially interact with alpha 2 receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibitory effects of osemozotan, a serotonin 1A-receptor agonist, on methamphetamine-induced c-Fos expression in prefrontal cortical neurons.

Psychostimulants induce hyperlocomotion in normal subjects, although, they are effective in producing a calming effect in hyperactive subjects. This paradoxical effect has been related to changes in serotonin (5-HT) neurotransmission in hyperactive dopamine transporter-knockout mice. In addition, we observed that hyperlocomotion in mice lacking pituitary adenylate cyclase-activating polypeptide was attenuated by amphetamine dependent on 5-HT(1A) receptor signaling and that amphetamine, when co-administered with a 5-HT(1A) agonist, produced a calming effect in wild-type mice. Here, in an attempt to address how 5-HT(1A) receptor signaling exerts the calming action of psychostimulants, we examined c-Fos expression in several brain regions after administration of methamphetamine and osemozotan, a selective 5-HT(1A) receptor agonist. The number of c-Fos-positive cells was increased in the medial prefrontal cortex, striatum and nucleus accumbens in methamphetamine (3 mg/kg body weight)-injected mice. Osemozotan (1 mg/kg) significantly reduced the methamphetamine-induced c-Fos expression in the medial prefrontal cortex and striatum, but not in the nucleus accumbens. This osemozotan action was completely blocked by the 5-HT(1A) receptor antagonist WAY-100635 (1 mg/kg). As the prefrontal cortex is considered to be involved in the beneficial actions of psychostimulant medications for attention-deficit/hyperactivity disorder, the present result showing 5-HT(1A)-mediated inhibition of corticostriatal activity may partly be related to this psychostimulant action.

Inhibition of R 28935- and R 29814-induced hypotension by prazosin in anaesthetized normotensive rats.

The intravenous administration of erythro 1-(1-[2-(1,4-benzodioxane-2-yl)-2-hydroxyethyl]-4-piperidyl)-2-benzimidazolinone R 28935) and the threo from R 29814 to anaesthetized normotensive rats showed a dose-dependent decrease in mean arterial pressure. Previous (-1 hr) intraperitoneal (0.1 mg/kg) as well as subcutaneous (0.03 mg/kg) treatment with the alpha 1-adrenoceptor blocking drug prazosin antagonized the hypotensive responses to R 28935 and R 29814. The selective antagonist of alpha 2-adrenoceptors yohimbine (0.5 mg/kg) was ineffective. On the other hand, the hypotensive action of clonidine was hardly affected by this prazosin treatment, whereas yohimbine now significantly impaired it. R 28935 and R 29814 showed no direct alpha-adrenoceptor stimulating properties and were moderately active in inhibiting the pressor response to (-)-phenylephrine in pithed normotensive rats when compared with phentolamine and prazosin. The results confirm and extend previous findings in cats. It is concluded that in some way central alpha 1-adrenoceptors are involved in the hypotensive mechanism of R 28935 (R 28914). However, central alpha 1-adrenoceptors are probably not the common sites of interaction, since any alpha 1-adrenoceptor-stimulating potency is lacking for R 28935 (R 29814) and their (moderate) affinity for alpha 1-adrenoceptors bears no relationship to their hypotensive activity.

Human plasma-derived material with clonidine displacing substance (CDS)-like properties contracts the isolated rat aorta.

1. The biological activity of a plasma-derived, clonidine displacing substance (CDS)-like material was tested on isolated rat aortic rings and compared to that of clonidine, an imidazoline with alpha 2-adrenoceptor agonist properties. 2. The CDS-like material was partially purified from expired human blood. This product inhibited in a dose-dependent manner the binding of [3H]-clonidine to rat brain membranes with a ki of 0.87 +/- 0.4 u ml-1, without affecting the binding of the alpha 1-antagonist, [3H]-WB4101. 3. When the CDS-like material (0.14-6 u ml-1) was applied to the bathing medium of isolated rat aortic rings, it caused dose-dependent contractions with an EC50 of 1.0 +/- 0.18 u ml-1. Clonidine also dose-dependently contracted rat aortic rings (EC50, 1.1 +/- 0.24 x 10(-7) M). The maximal tension developed in response to clonidine, however, was higher (1.37 +/- 0.15 g) compared to that developed in response to the CDS-like material (0.92 +/- 0.12 g). 4. Contractions induced by both CDS-like material and clonidine were antagonized by 5 x 10(-7) M rauwolscine, an alpha 2-adrenoceptor antagonist. Prazosin, an alpha 1-adrenoceptor antagonist, at 10(-8) M, greatly reduced contractions caused by clonidine while leaving those caused by CDS-like material unaffected. 5. The CDS-like material failed to alter the tension of intact or endothelium-denuded rat aortic rings which had been precontracted with methoxamine. Clonidine on the other hand, caused dose-dependent relaxations in intact, though not in denuded, precontracted rat aortic rings.(ABSTRACT TRUNCATED AT 250 WORDS)

Central 5-HT1A receptors and the mechanism of the central hypotensive effect of (+)8-OH-DPAT, DP-5-CT, R28935, and urapidil.

This study investigated the central hypotensive effects of drugs that possess a high affinity for central 5-hydroxytryptamine (5-HT1A) binding sites; (+)8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), N,N-dipropylcarboxamidotryptamine (DP-5-CT), erythro-1-(1-[2(1,4-benzodioxan-2-yl)-2-hydroxyethyl]- 4-piperidyl)-2-benzimidazolinone (R28935) and urapidil proved to possess high affinity and selectivity for central 5-HT1A binding sites, labeled by [3H]8-OH-DPAT. (+)8-OH-DPAT (0.1-10 micrograms/kg) given through the left vertebral artery of chloralose-anesthetized cats, lowered blood pressure by a biphasic dose-response curve. When given systemically, 10- to 100-fold higher doses of (+)8-OH-DPAT were necessary to obtain the same hypotensive effect when compared with central administration. Besides 8-OH-DPAT, R28935, DP-5-CT and urapidil also lowered blood pressure by a central mechanism in doses that were ineffective when given systemically. The central hypotensive effect of 0.3 micrograms/kg (+)8-OH-DPAT, 3 micrograms/kg DP-5 CT, and 3 micrograms/kg R28935 could be blocked by 100 micrograms/kg (-)pindolol, indicating that central 5-HT1A receptors are involved. High doses of (+)8-OH-DPAT (3-10 micrograms/kg) can also lower blood pressure by activating central alpha 2-adrenoceptors. The hypotensive effect of 300 micrograms/kg urapidil given through the vertebral artery could not be blocked by (-)pindolol. These results indicate the involvement of central 5-HT1A receptors in the mechanism of the central hypotensive activity of (+)8-OH-DPAT, DP-5-CT, and R28935.