Presence Of Phentolamine Research Articles

Comparison of Neurotransmitter Release from Perivascular Nerves in C57BL/6J Mice and Wistar Rats

It is thought that sympathetic nerve activation stimulates arterial constriction predominately through the release of three neurotransmitters: norepinephrine (NE), adenosine triphosphate (ATP) and neuropeptide Y (NPY). The aim of the current study was to examine the composition of neurotransmitter released in response to sympathetic nerve stimulation in mesenteric arteries of two commonly used wild‐type animal models. Isolated mesenteric arteries from male C57BL/6J mice and Wistar rats were mounted in a Danish Myotechnology 2‐Channel Wire Myograph System and isometric force measurements were recorded. Electrical field stimulation (EFS) was used to stimulate sympathetic perivascular nerves (PVNs) to release neurotransmitters in the presence and absence of the adrenergic receptor blocker phentolamine and/or purinergic receptor blocker NF449. Phentolamine alone blocked a significant proportion of the contractile response evoked by EFS in both mice and rats, suggesting that NE is the main sympathetic neurotransmitter in both species. Treatment with NF449 alone blocked a small proportion of the contractile response evoked by EFS in mice but did not have a significant effect in rat, suggesting that ATP may play a more important role in mice. The application of both phentolamine and NF449 nearly completely blocked the contractile response to EFS in mouse mesenteric arteries; however, in rat, a proportion of the contractile response, roughly equivalent to the response obtained in the presence of phentolamine alone, was still observed. These data suggest that the neurotransmitter composition is different between mice and rats, in particular mouse mesenteric sympathetic PVNs release NE and ATP while rat mesenteric sympathetic PVNs release NE and another non‐purinergic neurotransmitter(s).

A Role for Adrenergic Receptors in the Uterotonic Effects of Ergometrine in Isolated Human Term Nonlaboring Myometrium

Ergometrine is a uterotonic agent that is recommended in the prevention and management of postpartum hemorrhage. Despite its long-standing use, the mechanism by which it acts in humans has never been elucidated fully. The objective of this study was to investigate the role of adrenoreceptors in ergometrine's mechanism of action in human myometrium. The study examined the hypothesis that α-adrenoreceptor antagonism would result in the reversal of the uterotonic effects of ergometrine. Myometrial samples were obtained from women undergoing elective cesarean delivery. The samples were then dissected into strips and mounted in organ bath chambers. After the generation of an ergometrine concentration-response curve (10 to 10 M), strips were treated with increasing concentrations of ergometrine (10 to 10 M) alone and ergometrine (10 to 10 M) in the presence of phentolamine (10 M), prazosin (10 M), propranolol (10 M), or yohimbine (10 M). The effects of adding ergometrine and the effect of drug combinations were analyzed using linear mixed effects models with measures of amplitude (g), frequency (contractions/10 min), and motility index (g×contractions/10 min). A total of 157 experiments were completed on samples obtained from 33 women. There was a significant increase in the motility index (adding 0.342 g × counts/10 min/μM; 95% confidence interval [CI], 0.253-0.431, P < .001), amplitude (0.078 g/μM; 95% CI, 0.0344-0.121, P = 5e-04), and frequency (0.051 counts/10 min/μM; 95% CI, 0.038-0.063, P < .001) in the presence of ergometrine. The α-adrenergic antagonist phentolamine and the more selective α1-adrenergic antagonist prazosin inhibited the ergometrine mediated increase in motility index, amplitude, and frequency (-1.63 g × counts/10 min/μM and -16.70 g × counts/10 min/μM for motility index, respectively). These results provide novel evidence for a role for α-adrenergic signaling mechanisms in the action of ergometrine on human myometrial smooth muscle in the in vitro setting. Information that sheds light on the mechanism of action of ergometrine may have implications for the development of further uterotonic agents.

The characteristics of genistin-induced inhibitory effects on intestinal motility

Genistin belongs to isoflavones. Based on the facts that genistin exerts inhibitory effects on the contractility of vascular smooth muscle,the present study was designed to characterize the effects of genistin on intestinal contractility and evaluate its potential clinical implication. Ex vivo [isolated jejunal segment (IJS) of rat], in vitro, and in vivo assays were used in the study. The results indicated that genistin (5-80 μmol/L) inhibited the contraction of IJS in a dose-dependent manner and inhibited the increased-contractility of IJS induced by acetylcholine (ACh), histamine, high Ca(2+), and erythromycin, respectively. The inhibitory effects of genistin were correlated with the stimulation of alpha adrenergic and beta adrenergic receptors since these inhibitory effects were significantly blocked in the presence of phentolamine and propranolol respectively. No further inhibitory effects of genistin were observed in the presence of verapamil or in Ca(2+)-free condition, indicating genistin-induced inhibitory effects are Ca(2+)-dependent. Genistin decreased myosin light chain kinase (MLCK) protein contents and MLCK mRNA expression in IJS, and inhibited both phosphorylation and Mg(2+)-ATPase activity of purified myosin, implicating that the decrease of MLCK contents and inhibition of MLCK activity are involved in the genistin-induced inhibitory effects. The study suggests the potential clinical implications of genistin in relieving intestinal hypercontractility.

Characterization of beta-adrenoceptors mediating relaxation of the guinea-pig ileum.

Relaxation responses to sympathomimetic amines were recorded in potassium-contracted segments of guinea-pig ileum. Experiments were performed in the presence of phentolamine (5 X 10(-6) M) to eliminate beta-adrenoceptor-mediated effects. Metanephrine (10(-5) M) and desmethylimipramine (5 X 10(-7) M) were also present to prevent extraneuronal and neuronal uptake respectively. A potency order (-)-isoprenaline greater than (-)-noradrenaline greater than (-)-adrenaline was established, indicating a beta 1-adrenoceptor involvement for this relaxation. The potency of salbutamol (beta 2-selective) relative to isoprenaline in the ileum compared closely with its relative potency in isolated cardiac tissues (beta 1) but differed significantly from the value in lung parenchymal strips and vas deferens (beta 2). The pA2 values for antagonism of selective agonists (-)-noradrenaline (beta 1-selective) and fenoterol (beta 2-selective) by practolol (beta 1-selective) were identical, indicating a single beta 1-adrenoceptor population. The pA2 values for antagonism of these agonists by ICI 118,551 (beta 2-selective) were also identical and compatible with a beta 1-adrenoceptor population. Relaxation of the guinea-pig ileum is therefore mediated via a homogeneous population of beta 1-adrenoceptors.

Noradrenergic component in the vasoconstriction induced by 5-hydroxytryptamine in goat cerebral arteries.

5-Hydroxytryptamine (5-HT) elicited dose-dependent increases in tension in the middle cerebral artery of the goat, which were significantly antagonized by lysergic acid diethylamide (LSD, 10(-8) M) and methysergide (10(-7) M). In the presence of phentolamine (10(-6) M), the dose-response curve to 5-HT was shifted to the right, the pA2 value for this antagonism was 6.52. Pretreatment of goats with reserpine (0.02 mg kg-1 day-1 for three days) or removal of both superior cervical ganglia 15 days before the experiment brought about a significant decrease in the vasoconstriction induced by doses of 5-HT higher than 10(-7) M. The remaining contraction produced by 5-HT in arterial segments from reserpinized or gangliectomized goats was further reduced in the presence of LSD. In addition, high concentrations of 5-HT induced tritium release from goat pial arteries preloaded with (-)-[3H]noradrenaline, 2 X 10(-7) M) which was significantly decreased in vessels from gangliectomized or reserpinized goats. These results in goat cerebral arteries indicate that in the contraction evoked by 5-HT there are two components. The first appears with low concentrations (up to 10(-7) M) in which 5-HT acts directly on 5-HT receptors. The second occurs at high doses (greater than 10(-7) M) in which 5-HT also acts indirectly on alpha-adrenoceptors by release of noradrenaline from noradrenergic nerve endings.

Actions of dopamine and apomorphine on the vasoconstrictor responses of perfused mesenteric arteries of mouse, rat and rabbit.

Dopamine (10(-7)--10(-6) M) and apomorphine (5 x 10(-7)--5 x 10(-6) M) inhibited the vasoconstrictor responses of the perfused mesenteric artery preparations of rat, rabbit and mouse to adrenergic nerve stimulation but did not affect responses to added noradrenaline. The inhibitory effects of both dopamine and apomorphine were prevented by haloperidol (3 x 10(-7) M) but not by yohimbine (3 x 10(-8) M) in rat and rabbit mesenteric artery preparations. In contrast, yohimbine (3 x 10(-8) M), but not haloperidol, antagonized the inhibitory effect of dopamine and apomorphine in mouse mesenteric artery preparations. In higher concentrations, dopamine (10(-6)--10(-4) M) produced a direct vascoconstrictor effect, which involved post-junctional alpha-adrenoceptors in all three species. However, in preparations contracted with 10(-7) M 5-hydroxytryptamine and in the presence of phentolamine (3 x 10(-7) M) and propranolol (10(-6) M), dopamine (10(-6)--10(-4) M) produced a direct relaxant effect in rabbit mesenteric artery preparations but not in those of rat and mouse. It is suggested that inhibition of neurogenic vasoconstrictor responses, by dopamine and apomorphine, may be mediated through a specific prejunctional inhibitory dopamine receptor in the mesenteric artery of rat and rabbit whereas in the mouse they involve activation of alpha-adrenoceptors.

Octopamine—A single modulator with double action on the heart of two insect species ( Apis mellifera macedonica and Bactrocera oleae): Acceleration vs. inhibition

The effects of octopamine, the main cardioacceleratory transmitter in insects, were investigated, in the isolated hearts of the honeybee, Apis mellifera macedonica, and the olive fruit fly, Bactrocera oleae. Octopamine induced a biphasic effect on the frequency and force of cardiac contractions acting as an agonist, with a strong acceleratory effect, at concentrations higher than 10 −12 M for the honeybee and higher than 50 × 10 −9 M for the olive fruit fly. The heart of the honeybee is far more sensitive than the heart of olive fruit fly. This unusual sensitivity is extended to the blockers of octopaminergic receptors, where phentolamine at 10 −5 M stopped the spontaneous contractions of the honeybee heart completely and permanently, while the same blocker at the same concentration caused only 50% inhibition in the heart of the olive fruit fly. Phentolamine and mianserin at low concentrations of 10 −7 M also blocked the heart octopaminergic receptors, but for a short period of time, of less than 15.0 min, while a partial recovery in heart contraction started in spite of the presence of the antagonist. The unusual response of the honeybee heart in the presence of phentolamine and/or mianserin suggests excitatory effects of octopamine via two different receptor subtypes. At lower concentrations, 10 −14 M, the agonist octopamine was converted to an antagonist, inducing a hyperpolarization in the membrane potential of the honeybee cardiac pacemaker cells and inhibiting the firing rate of the heart. The inhibitory effects of octopamine on certain parameters of the rhythmic bursts of the heart of the honeybee, were similar to those of mianserin and phentolamine, typical blockers of octopaminergic receptors. The heart of the olive fruit fly was 10 5 times less sensitive to octopamine, since a persistent inhibition of heart contractions occurred at 10 −9 M. In conclusion, the acceleration of the insect heart is achieved by increasing the levels of octopamine, while there is a passive but also an active decrease in heart activity due to the minimization of octopamine.

In vitro determination of the mechanism of the uterine stimulatory effect of Newbouldia laevis

The uterine stimulatory effect of the ethanol leaf extract of Newbouldia laevis (Beauv.) Seemann ex Bureau (Bignoniaceae) was evaluated in the presence of some antagonists in vitro in an attempt to elucidate the mechanism of action of the extract. The extract was tested in the presence and absence of phentolamine (4.09 and 40.91 nM), diphenhydramine (4.45 and 44.47 nM), atropine (1.18 and 11.91 nM), and verapamil (2.03 and 20.35 nM). The effect of the antagonists on the extract and on oxytocin used as a reference drug in this study was evaluated. The EC50 and Emax were determined and statistically analyzed using one way ANOVA and Dunnett’s post hoc test. There was no significant difference in the EC50 and Emax of the extract and oxytocin in the presence of phentolamine. Diphenhydramine and atropine significantly inhibited (p <0.01) the extract but both drugs had no effect on oxytocin. However, significant differences (p <0.01) were observed in the EC50 and Emax of the extract and oxytocin in the presence of verapamil. These results suggest that the leaf extract of N. laevis contracts the uterus by opening voltage-operated calcium channels and/or by activation of muscarinic receptors.

Norepinephrine, via β-Adrenoceptors, Regulates Bumetanide-Sensitive Cotransporter Type 1 Expression in Thick Ascending Limb Cells

The sympathetic nervous system, via norepinephrine, regulates renal sodium transport, and chronic sympathetic activation causes sustained increases in blood pressure by reducing sodium excretion. Our previous studies show that chronic norepinephrine infusion increases the abundance of the bumetanide-sensitive cotransporter type 1, the apical sodium transporter of the thick ascending limb of Henle's loop. The present study was initiated to elucidate the mechanisms by which norepinephrine regulates the protein levels of this transporter in an immortalized thick ascending limb epithelial cell line. Treatment with norepinephrine, either alone or in the presence of actinomycin D or cycloheximide, had no effect on cotransporter mRNA levels. Treatment with norepinephrine, however, increased bumetanide-sensitive cotransporter type 1 protein levels (70% increase versus control; P=0.012), and pretreatment with cycloheximide blocked the effect of norepinephrine on bumetanide-sensitive cotransporter type 1 protein levels. To further elucidate the mechanism, thick ascending limb cells were treated with norepinephrine in the presence of phentolamine (alpha-adrenoceptor blocker), propranolol (beta-adrenoceptor blocker), SQ22536 (adenylyl cyclase inhibitor), PD098059 (mitogen-activated protein kinase pathway inhibitor), H-89 (protein kinase A inhibitor), or staurosporine (protein kinase C inhibitor). Treatment with propranolol, SQ22536, and H-89 abolished the effects of norepinephrine on bumetanide-sensitive cotransporter type 1 protein levels, whereas staurosporine had no effect. Treatment with PD098059 partially inhibited the effects of norepinephrine (40% decrease versus norepinephrine; P=0.03), and treatment with phentolamine potentiated the effects of norepinephrine (30% increase versus norepinephrine; P=0.02) on bumetanide-sensitive cotransporter type 1 protein levels. We conclude that regulation of bumetanide-sensitive cotransporter type 1 by norepinephrine proceeds via the beta-adrenoceptor receptor-cAMP-protein kinase A pathway that involves in part mitogen-activated protein kinases and that alpha-adrenoceptor activation negatively regulates bumetanide-sensitive cotransporter type 1 protein levels.

Pharmacological characterization of the β-adrenoceptor that mediates the relaxant response to noradrenaline in guinea-pig tracheal smooth muscle

Pharmacological characteristics of beta-adrenoceptors (beta-ARs) mediating noradrenaline-induced relaxation were investigated in guinea-pig tracheal smooth muscle. The inhibitory effects of several types of beta-AR antagonists on noradrenaline-induced relaxation against histamine contraction were scrutinized with Schild plot analysis. The concentration-response curve for noradrenaline obtained in the absence of phentolamine and uptake inhibitors was competitively antagonized by all of the beta-AR antagonists used in this study (propranolol, bupranolol, atenolol, butoxamine and ICI-118,551). However, their pA2 values were markedly less than the expected values for beta1-AR and beta2-AR. On the other hand, pA2 values of ICI-118,551 (6.85) determined in the presence of phentolamine suggested a contribution of a beta1 -AR rather than beta2 -AR. In the presence of phentolamine and uptake inhibitors (desipramine and deoxycorticosterone), the Schild plot for atenolol was a better fit, with two distinct straight lines. The pA2 values of atenolol provided by the regression were: approximately 7.0, which corresponds to the expected beta1-AR value, and approximately 6.5, which was 3 times less than the expected value for beta1 -AR, and thus the possible presence of two classes of beta1 -AR (beta1(Low) and beta1(High)) was suggested. This view was also supported by Schild plot analysis for propranolol, which fit two straight lines each with a slope of 1.0. The present findings indicate that beta1 -ARs contributing to noradrenaline-elicited relaxation in guinea-pig tracheal smooth muscle exhibit diverse pharmacological characteristics and may be subdivided into at least two classes with distinct affinities for atenolol. Under physiological conditions, beta1(Low) rather than beta1(High) seems to play a more significant role in noradrenaline-regulated airway smooth muscle tone.

.ALPHA.-Adrenergic Receptor Mediated Hypertensive and Vasoconstrictor Effects of Dietary Radish Leaves Extract

Radish is a universally known food plant. Leaves of radish are widely used for their culinary and medicinal properties. In this study we report the hypertensive and vasoconstrictor effects of the aqueous crude extract of radish leaves (Rl.Cr )m ediated possibly via activation of α-adrenergic receptors. Rl.Cr, that tested positive for the presence of saponins and alkaloids, exhibited a dose-dependent (10–300 mg/kg) hypertensive effect in the blood pressure (BP) of rats under anaesthesia. This effect was blocked in the presence of phentolamine (2 mg/kg), similar to norepinephrine (NE) a standard adrenergic agonist. Further investigations were conducted on the isolated tissue preparations. In rabbit aorta, Rl.Cr showed a dose-dependent (0.3–5 mg/ml) vasoconstrictor effect that was sensitive to phentolamine pretreatment. Phentolamine pretreatment also shifted the curves of Rl.Cr in a parallel manner to the right in aorta, similar to NE, thus possibly confirming the adrenergic receptor mediated effect. However, Rl.Cr was devoid of any activity in isolated guinea pig atria up to the dose of 10 mg/ml indicating that the extract has a specific α-adrenergic effect independent of β-adrenergic receptor involvement. Thus the study shows hypertensive and vasoconstrictor activities of radish leaves extract possibly mediated via interaction at α-adrenergic receptors.

Catecholaminergic activation of G-protein coupling in rat spinal cord: Further evidence for the existence of dopamine and noradrenaline receptors in spinal grey and white matter

[ 35S]GTPγS autoradiography of slide-mounted tissue sections was used to examine G-protein coupling in the rat spinal cord, as stimulated by dopamine, the D1 receptor agonist SKF 38393, noradrenaline, and noradrenaline in the presence of the alpha adrenoceptor antagonist, phentolamine. Measurements were obtained from the different laminae of spinal grey and from the dorsal, lateral, and ventral columns of white matter, at cervical, thoracic, and lumbar levels. At every level, there was a relatively strong basal incorporation of GTPγS in laminae II–III > lamina IV–X of spinal grey, even in presence of DPCPX to block endogenous activation by adenosine A1 receptors. Dopamine, and to a lesser degree SKF 38393, but not the D2 receptor agonist quinpirole, stimulated G-protein coupling in laminae IV–X. Both dopamine and SKF 38393 also induced a weak but significant activation throughout the white matter. In both grey and white matter, the activation by dopamine was markedly reduced in presence of a selective D1 receptor antagonist. Noradrenaline strongly stimulated coupling throughout the spinal grey at all levels, an effect that was uniformly reduced in the presence of phentolamine. With or without phentolamine, there was also significant stimulation by noradrenaline in the white matter. Under the same experimental conditions, alpha 1, alpha 2, and beta adrenergic receptor agonists failed to activate GTPγS incorporation in either grey or white matter. However, in the presence of selective alpha 1 or alpha 2 receptor antagonist, significant reductions of noradrenaline-stimulated GTPγS incorporation were observed in both grey and white matter. The beta antagonist propanolol reduced GTPγS incorporation in grey matter only. Thus, the results confirmed the existence of D1 dopamine receptors and of alpha 1, alpha 2, and beta adrenergic receptors in the grey matter of rat spinal cord. In white matter, they strongly suggested the presence of dopamine D1, and of alpha 1 and alpha 2 adrenergic receptors on glia and/or microvessels, that might be activated by diffuse transmission in vivo.

ATP‐ and adenosine‐induced relaxation of the smooth muscle of the pig urethra

To investigate relaxation mechanisms for ATP and adenosine in the pig urethra, together with the possible role of ATP in nerve-evoked urethral relaxations, as ATP is thought to cause bladder smooth muscle contraction via P2X receptors, whereas relaxation is mediated via G-protein coupled P2Y receptors, and ATP may also induce relaxation via breakdown to adenosine. Circular muscle strips from the female pig urethra were mounted in tissue baths to record force; the effects of increasing concentrations of 1-300 microM ATP, the P2-receptor agonist 2-methylthioATP (2-MeSATP), adenosine, the stable adenosine-analogue, 5'(N-ethylcarboxamido) adenosine (NECA), ADP, uridine-triphosphate (UTP) and alpha,beta-methylene-ATP were assessed on the spontaneously developed tone. Responses to ATP were further assessed in the presence of G-protein activator guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS; 1-10 microM), the G-protein inhibitor guanosine 5'-O-(2-thio-diphosphate) (GDPbetaS; 10-100 microM), suramin (1-100 microM), the ecto-ATPase inhibitor 6-N,N-diethyl-beta-gamma-dibromomethylene-D-adenosine-5-triphosphate (ARL 67156, 10-100 microM), and the suggested P2Y receptor antagonist, reactive blue-2 (1-100 microM). The effect of the adenosine (P1) receptor antagonist 8-(p-sulphophenyl)theophylline (8-SPT, 1-100 microM) on responses to adenosine, and the effects of the adenosine reuptake inhibitor S(p-nitrobenzyl)-6-thioinosine (NBTI, 1-100 microM) on responses to adenosine and ATP were also assessed. Responses to electrical field stimulation (EFS, 12 and 30 Hz) in the presence of phentolamine (1 microM), scopolamine (1 microM) and N omega-nitro-L-arginine (0.3 mM) were studied before and after treatment with GTPgammaS, GDPbetaS, suramin, reactive blue-2 and ARL 67156. Strips were relaxed in a concentration-dependent manner by exogenously administered ATP and 2-meSATP, the relaxations being slowly developing and long-lasting. The relaxant effect evoked by both agonists at 300 microM amounted to about half of the spontaneously developed tone. The relaxation evoked by ATP was not significantly affected by GTPgammaS, GDPbetaS, suramin, ARL 67156 or reactive blue-2. Adenosine induced a concentration-dependent relaxation of the smooth muscle tone, reaching a maximum of approximately 70% at 300 microM, whereas 300 microM NECA only relaxed the preparations by approximately 35%. The adenosine-induced relaxation was not affected by treatment with 8-SPT. However, NBTI (1 microM) significantly reduced the relaxation evoked by 300 microM adenosine. ADP relaxed the smooth muscle tone by approximately 40% (300 microM). There was no response to UTP, and the effect of alpha,beta-methylene-ATP was negligible (5% relaxation at 100 microM). EFS caused slowly developing and long-lasting relaxations that were unaffected by GTPgammaS, GDPbetaS, suramin, reactive blue-2 and ARL 67156. These results suggest that exogenous ATP and adenosine relax the smooth muscle of the pig urethra in a manner similar to that evoked by electrical stimulation of nerves, although there was no evidence for involvement of a definable P2Y receptor subtype in these relaxations.

Comparaison de l’activité réflexe de l’utérus au cours de l’insémination artificielle et de l’accouplement chez la brebis

To identify and to compare the reflex mechanisms involved in the activation of the uterine motility following artificial insemination (AI) and mating in the ewe. The contractile activity of the uterus was monitored in presence and absence of adrenergic (phentolamine) and cholinergic (atropine) antagonists, using an implantable telemetric device equipped with a sensor catheter inserted into the uterine lumen, and connected to a radio-telemetric transmitter. During AI, a uterine contraction (UC) of short duration (<20 seconds) appeared in direct response to animal restraint, to the speculum introduction, then when the speculum was opened. Both the UC evoked by restraint of the ewe and insertion of the speculum were abolished in presence of phentolamine, while atropine inhibited the UC in response to opening of the speculum only. After AI, the uterine activity increased sharply and was all the more intense and extended with higher vaginal wall pressure. Phentolamine or atropine did not inhibit this motor response, whereas a similar pattern of uterine hyperactivity occurred following i.v. injection of oxytocin (100 and 200 mIU; Syntocinon). After mating, an increase in uterine activity was never observed. Only penis intromission evoked a UC of short duration (about 20 seconds), abolished in presence of phentolamine only. Artificial insemination in the ewe increases uterine motility, resulting from the reflex activation of adrenergic and cholinergic nerve fibres of the autonomic nervous system, following by a reflex release of oxytocin from the pituitary gland (also called "Ferguson reflex"). This secretion of OT was elicited by the excessive dilation of the vaginal wall with the speculum. By comparison, mating did not evoke a period of uterine hyperactivity and respects the physiological post-coital resting period.

Glucose Dependence of Imidazoline-Induced Insulin Secretion

The glucose dependence of the insulinotropic action of KATP channel-blocking imidazoline compounds was investigated. Administration of 100 micromol/l phentolamine, but not 100 micromol/l efaroxan, markedly increased insulin secretion of freshly isolated mouse islets when the perifusion medium contained 5 mmol/l glucose. When the glucose concentration was raised to 10 mmol/l in the continued presence of either imidazoline, a clear potentiation of secretion occurred as compared with 10 mmol/l glucose alone. In the presence of efaroxan, a brisk first-phase-like increase was followed by a sustained phase, whereas a more gradual increase resulted in the presence of phentolamine. Administration of 100 micromol/l phentolamine was somewhat more effective than 100 micromol/l efaroxan to inhibit KATP channel activity in intact cultured beta-cells (reduction by 96 vs. 83%). Both compounds were similarly effective to depolarize the beta-cells. When measured by the perforated patch-technique, the depolarization by efaroxan was often oscillatory, whereas that by phentolamine was sustained. In perifused cultured islets, both compounds increased the cytosolic calcium concentration ([Ca2+]c) in the presence of 5 and 10 mmol/l glucose. Efaroxan induced large amplitude oscillations of [Ca2+]c, whereas phentolamine induced a sustained increase. It appears that a KATP channel block by imidazolines is not incompatible with a glucose-selective enhancement of insulin secretion. The glucose selectivity of efaroxan may involve an inhibitory effect distal to [Ca2+]c increase and/or the generation of [Ca2+]c oscillations.

Acute cardiovascular effects of sibutramine in conscious rats.

Sibutramine is a serotonin and norepinephrine reuptake inhibitor, used in the treatment of obesity. In this study, cardiovascular effects of sibutramine (0.9, 3, or 9 mg kg(-1) i.p.) were measured in conscious Sprague-Dawley rats, in the absence and presence of beta- and/or alpha-adrenoceptor antagonism (with propranolol and/or phentolamine, respectively). Sibutramine caused pressor and tachycardic effects, with celiac and mesenteric vasoconstrictions, and hyperemic hindquarters vasodilatation. Pretreatment with propranolol inhibited the tachycardic and hindquarters vasodilator effect of sibutramine, whereas phentolamine inhibited the pressor and vasoconstrictor effects of sibutramine. In the presence of phentolamine, sibutramine caused hyperemic mesenteric vasodilatation. In preconstricted, isolated, mesenteric vessels, sibutramine and its metabolites BTS 54505 (N-desmethylsibutramine) and BTS 54354 (N-didesmethylsibutramine) (10 microM) produced significant vasodilations. Neither sibutramine nor BTS 54505 enhanced vessel sensitivity to norepinephrine, whereas BTS 54 354 produced a significant leftward shift in the concentration-response curve to norepinephrine. Collectively, the results indicate that the overt cardiovascular effects of sibutramine involve alpha-adrenoceptor-mediated celiac and mesenteric vasoconstrictions, and beta-adrenoceptor-mediated hindquarters vasodilatation and tachycardia. The mesenteric vasodilator response to sibutramine, seen in the presence of phentolamine, may be a direct effect of the drug and/or its metabolites, on vessel tone. The cardiovascular effects of sibutramine in vivo may be secondary to inhibition of peripheral and/or central reuptake of monoamines by the metabolites BTS 54354 and/or BTS 54505. It remains to explain why BTS 54354, but not BTS 54505, enhanced norepinephrine sensitivity in vitro, because both metabolites are potent inhibitors of the norepinephrine transporter.

Control of non-adrenergic non-cholinergic reflex motor responses in circular muscle of guinea-pig small intestine by Met-enkephalin.

1 A triple organ bath method allowing the synchronous recording of the motor activity of the circular muscle layer belonging to the oral and anal segments of guinea-pig small intestine adjacent to an electrically stimulated middle segment was developed to study the ascending and descending reflex motor responses. 2 Electrical field stimulation (0.8 ms, 40 V, 5 Hz, 10 s) applied to the middle part of the segments elicited tetrodotoxin (1 microm)-sensitive ascending and descending contractile responses of the nonstimulated parts, oral and anal, respectively. The ascending contraction was more pronounced as compared with the descending contraction. 3 In the presence of phentolamine (5 microm), propranolol (5 microm) and atropine (3 microm) a significant decrease in the amplitude of the ascending contraction was seen and a descending relaxation, instead of a contraction was observed. 4 Met-enkephalin applied at a single concentration (0.1 microm) or cumulatively (0.001-1 microm) inhibited both non-adrenergic non-cholinergic (NANC) descending relaxation and ascending contraction with similar efficacy but different potency, IC50 being 5.9 +/- 0.3 and 39.0 +/- 4 nm, respectively. Naloxone (0.5 microm) prevented the effects of Met-enkephalin. 5 L-NNA (0.5 mm), an inhibitor of nitric oxide synthesis, increased the ascending contraction and strongly reduced but not abolished the descending relaxation. l-Arginine (0.5 mm) restored the motor responses to the initial level in l-NNA-pretreated preparations, d-Arginine (0.5 nm) had no effects. 6 Met-enkephalin (0.1 microm) depressed the l-NNA-dependent increase of the ascending contraction and failed to change the l-NNA-resistant part of the descending relaxation. 7 Met-enkephalin did not alter spontaneous NANC mechanical activity. SNP (1 or 10 microm), an exogenous donor of nitric oxide, caused a concentration-dependent relaxation. The effects of SNP persisted in Met-enkephalin (0.1 microm)-pretreated preparations. 8 NANC reflex ascending contraction and descending relaxation were synchronously induced by a local nerve stimulation indicating a functional coactivation of NANC orally projected excitatory and anally directed inhibitory pathways. Acting prejunctionally, Met-enkephalin provided a negative controlling mechanism inhibiting both ascending and descending, mainly nitric oxide mediated, reflex responses. A higher sensitivity of the descending relaxation to Met-enkephalin was observed suggesting an essential role of opioid(s) in reducing the efficacy of descending motor activity.

Noradrenaline-induced changes in intracellular Ca(2+) and tension in mesenteric arteries from diabetic rats.

1. The purpose of this investigation was to determine whether enhanced contractile responses to noradrenaline (NA) of mesenteric arteries from rats with chronic streptozotocin-induced diabetes are associated with increases in mean cytosolic [Ca(2+)]i. 2. [Ca(2+)]i was measured with fura 2-AM, and was monitored simultaneously with tension in perfused endothelium-denuded mesenteric arterial rings from 12 - 14 week diabetic rats and age- and gender-matched control rats. 3. Basal [Ca(2+)]i (expressed as R(n), the normalized fura 2 ratio) was not significantly different in arteries from control and diabetic rats. Similarly, no differences between control and diabetic arteries in the tension or [Ca(2+)]i responses to 80 mM KCl in the presence of phentolamine were detected. 4. The rate of tension development, peak tension and integrated tension in response to 30 microM NA were all significantly greater in diabetic than control arteries. However, this was not associated with enhancement of the corresponding [Ca(2+)]i responses in the diabetic arteries. 5. Peak contractile responses to perfusion with both 0.3 and 3 microM NA, but peak [Ca(2+)]i only in response to 0.3 microM NA, were significantly greater in diabetic than control arteries. 6. NA (30 microM) produced a greater increase in both peak tension and [Ca(2+)]i in diabetic than control arteries perfused with Ca(2+)-free solution containing 1 mM EGTA. Neither the rate nor the magnitude of NA-induced Ca(2+) influx appeared to be altered in the diabetic arteries. 7. The enhanced sustained contractile response of diabetic arteries to NA appears to be dissociated from increases in [Ca(2+)]i, and may be due to other factors, such as an increase in the Ca(2+) sensitivity of the contractile proteins.

Nisoxetine infusion into the olfactory bulb enhances the capacity for male rats to identify conspecifics.

In the present report, the norepinephrine uptake inhibitor nisoxetine as well as a cocktail of nisoxetine and the alpha-adrenergic receptor antagonist phentolamine were infused unilaterally into the olfactory bulb during microdialysis to assess their effects upon the capacity of male rats to identify conspecifics. A social discrimination test was conducted while simultaneously measuring olfactory bulb norepinephrine output in the dialysate before, during, and after behavioral testing. Nisoxetine significantly increased norepinephrine levels in the olfactory bulb compared with the Ringer's solution control group. Following such increases in olfactory bulb norepinephrine, identification responses were enhanced compared with that observed in the Ringer's control. In the presence of phentolamine, nisoxetine elevated olfactory bulb norepinephrine to levels similar to that obtained in the nisoxetine alone group, however, investigatory responses directed to the conspecifics indicated an absence of identification capacity similar to that observed in the Ringer's control group. These results reveal a direct link between norepinephrine transmission in the olfactory bulb and enhanced identification via its activation of postsynaptic alpha-adrenergic receptors. These results also show that inhibition of norepinephrine uptake may represent an important mechanism involved with the enhancement of social identification and suggest a possible novel effect for the antidepressant nisoxetine.

Acetylcholine-induced positive inotropy mediated by prostaglandin released from endocardial endothelium in mouse left atrium.

The possible involvement of the endocardial endothelium in the positive inotropic response of the mouse left atrium to acetylcholine was examined pharmacologically. In mouse left atria, acetylcholine produced a biphasic inotropic response: a transient decrease in contractile force followed by a late increase. The positive response was not affected by the presence of phentolamine and propranolol, but was almost abolished by pretreatment of the preparation with 1% Triton X-100, which denudes the endocardium of its endothelium. Nordihydroguaiaretic acid, NG-nitro-L-arginine, BQ-123 and BQ-788 had no effect on the inotropic responses to acetylcholine, but indomethacin completely abolished the positive response. Prostaglandins and their analogues had a positive inotropic effect with a potency order PGF2alpha>PGD2>PGE2>U46619, whereas beraprost had no effect. Neither Triton X-100 pretreatment nor the presence of indomethacin affected the positive inotropic effect of PGF2alpha. Acetylcholine and PGF2alpha prolonged the action potential duration similarly. These results suggest that the acetylcholine-induced positive inotropic response in mouse left atria is mediated by prostaglandin released from the endocardial endothelium.