N-NITRO-L-ARGININE AND INDOMETHACIN DO NOT AFFECT ENDOTHELIN-INDUCED CONSTRICTION OF LARGE AND SMALL CORONARY ARTERIES IN THE ANAESTHETIZED GREYHOUND

• 1 The aim of this study was to investigate whether endothelin-l (ET-1)-induced constriction of large and small coronary arteries in the anaesthetized greyhound is modulated by the endogenous release of nitric oxide or prostanoids.
• 2 ET-1 (1–100 ng/kg) and the α₁-adrenoceptor agonist phenylephrine (0.5-2 pg/kg), when injected directly into the circumflex coronary artery, caused dose-dependent decreases in epicardial coronary artery diameter and coronary vascular conductance without affecting systemic arterial pressure or the rate and force of cardiac contraction.
• 3 Inhibition of NO synthesis with N-nitro-L-arginine (NOLA, 5 mg/kg, i.c.) decreased coronary artery diameter, coronary conductance and heart rate and increased arterial pressure. The coronary vasoconstrictor response to ET-1 was unaffected by NOLA. By contrast, NOLA significantly increased the phenylephrine-induced constriction of the epicardial coronary artery but not the resistance vessels.
• 4 Indomethacin (5 mg/kg, i.v.), an inhibitor of cyclo-oxygenase, significantly decreased epicardial coronary artery diameter but did not affect coronary conductance. Indomethacin had no effect on the coronary vascular responses to ET-1 or phenylephrine. Combined treatment with NOLA plus indomethacin also failed to affect the coronary vasoconstrictor effects of ET-1.
• 5 Basal release of NO and vasodilator prostanoids modulated resting coronary vascular tone but did not Influence the vasoconstrictor responses to endothelin in either large or small coronary arteries.

     

THE CARDIOVASCULAR EFFECTS OF CENTRALLY ADMINISTERED 5-HYDROXYTRYPTAMINE IN THE CONSCIOUS NORMOTENSIVE AND HYPERTENSIVE RAT

• 1 The cardiovascular effects of centrally administered 5-hydroxytryptamine (5-HT) have been analysed in conscious normotensive and hypertensive rats.
• 2 In conscious normotensive rats, 5-HT, (1–30 μg) administered intracerebroventricularly (i.c.v.) produced profound and immediate dose-related decreases in heart rate and small increases in blood pressure. The initial pressor responses were followed by secondary secondary depressor responses at high doses of 5-HT.
• 3 Similar effects were produced by 5-HT i.c.v. in conscious DOCA-salt and spontaneously hypertensive rats, although the magnitude of the pressor responses was substantially greater in hypertensive than normotensive rats.
• 4 Pretreatment with either N-methylatropine or atenolol intra-arterially reduced the 5-HT-induced bradycardia in normotensive rats; the reduction was enhanced when both antagonists were given in combination.
• 5 The 5-HT₂ antagonist, cyproheptadine (10 μg i.c.v.) increased basal blood pressure and heart rate in normotensive rats. Subsequent administration of 5-HT i.c.v. produced biphasic effects on heart rate consisting of an initial tachycardia followed by a marked bradycardia.
• 6 Methysergide (10 μg i.c.v.) pretreatment did not alter resting heart rate, but attenuated the 5-HT induced bradycardia. A higher dose of methysergide, (30 μg i.c.v.), decreased resting blood pressure and heart rate.
• 7 This study has demonstrated, therefore, that the 5-HT induced bradycardia is produced by not only a centrally mediated decrease in sympathetic tone, but also an increase in vagal drive to the heart. The bradycardia is antagonised by centrally administered methysergide, but not by cyproheptadine, which suggests that it is probably mediated through a ‘5-HT₁-like’ receptor mechanism.

     

Femoral Vasodilation to Cromakalim is Blocked by U37883A,a Non-sulphonylurea that Selectively Inhibits KATP Channels

The purpose of the present study was to determine the effects of U37883A, a non-sulphonylurea inhibitor of K_ATP channels, in the femoral vascular bed of the anaesthetized dog. Administration of U37883A, 4-morpholinecarboxamidine-N-1-adamentyl-N-cyclohexyl hydrochloride (2·5 mg kg^?1, i.v.), significantly inhibited the femoral vasodilator response to intra-femoral arterial injection of cromakalim, an activator of K_ATP channels. In contrast, U37883A had no effect on the femoral vasodilator responses to nitroglycerin, isoprenaline, 5-HT, or 5-carboxamidotryptamine, suggesting this agent is a novel and selective inhibitor of hindlimb vasodilation induced by K_ATP-channel activation. Since U37883A did not significantly alter baseline femoral blood flow and femoral vascular resistance, the present data suggest that K_ATP channels do not contribute, in large measure, to regulating the canine femoral vascular bed under resting conditions in-vivo.     

Effects of methysergide, pizotifen and ergotamine in the monkey cranial circulation.

Internal and external carotid vascular resistances were measured, in anaesthetized monkeys, to asses the direct cranial vascular effects of i.v. methysergide, pizotifen and ergotamine, and their effects on the cranial vascular responses to the constrictors 5-hydroxytryptamine and noradrenaline and the dilators histamine, prostaglandin E1 and bradykinin. Methysergide reduced responses to 5-HT, and tended to potentiate the external carotid responses to noradrenaline. Pizotifen blocked responses to histamine; it tended to reduce internal carotid responses to 5-HT, but it potentiated external carotid 5-HT responses. Ergotamine reduced responses to 5-HT and noradrenaline, but this was probably related to its cranial vasoconstrictor effects, especially in the external carotid circulation. Methysergide induced weak transient cranial vasoconstriction and pizotifen had no direct effects. These findings may be relevant to the therapeutic actions of these drugs in migraine, since the doses used approximated to those used clinically.     

Pharmacological profile of 5-hydroxytryptamine-induced inhibition on the pressor effect elicited by sympathetic stimulation in long-term diabetic pithed rats

We analysed the type and/or subtype of 5-hydroxytryptamine (5-HT) receptors involved in the inhibitory mechanisms of 5-HT on the pressor responses induced by stimulation of sympathetic vasopressor outflow in long-term diabetic pithed rats. Diabetes was induced in male Wistar rats by a single subcutaneous injection of alloxan. Eight weeks later, rats were anaesthetized, pre-treated with atropine, and pithed. The effect of 5-HT on the pressor responses elicited by stimulation of the sympathetic outflow was analysed in eight-week alloxan-induced diabetic pithed rats. 5-HT (20 μg/kg/min) reduced the pressor action obtained by electrical stimulation of the sympathetic outflow. However, there was no effect on exogenous noradrenaline-induced pressor responses. 5-CT (5 μg/kg/min), 8-OH-DPAT (5 μg/kg/min), and α-methyl-5-HT (5 μg/kg/min), selective 5-HT₁, 5-HT_1A and 5-HT₂ receptor agonists, respectively, reproduced the 5-HT inhibitory action. Nevertheless, infusion of 5 μg/kg/min of 1-phenylbiguanide, CGS-12066B, L-694,247, BW273C86 or MK212 (5-HT₃, 5-HT_1B, 5-HT_1D, 5-HT_2B and 5-HT_2C receptor agonists, respectively) had no effect on the pressor responses elicited by stimulation of the sympathetic outflow. Methiothepin (100 μg/kg) and a cocktail of WAY-100,635 (100 μg/kg) and spiperone (125 μg/kg) blocked the 5-HT inhibitory effect on the pressor action obtained by sympathetic stimulation. Moreover, WAY-100, 635 abolished the 8-OH-DPAT inhibitory effect and spiperone blocked α-methyl-5-HT action. In conclusion, this study revealed that long-term experimental diabetes induces changes in the receptor type/subtype involved in the 5-HT inhibitory action on the sympathetic pressor responses produced by electrical stimulation. This is mainly mediated by pre-junctional 5-HT_1A and 5-HT_2A receptors.     

DUAL PRESYNAPTIC EFFECTS OF 5-HYDROXYTRYPTAMINE ON PERIPHERAL NORADRENERGIC SYNAPSES

• 1 The aim of the present experiments was to study the effects of 5-hydroxytryptamine (5-HT) on the responses to postganglionic stimulation of two models of the peripheral sympathetic nervous system: the isolated nictitating membrane of the cat and the guinea-pig isolated atria.
• 2 In the nictitating membrane of the cat, 5-HT (0.1 μM) shifted to the left the frequency-response curve to nerve stimulation. This potentiating effect was prevented by 5-HT receptor antagonists (0.1 μM. methysergide, 0.1 μM pizotifen and 0.1 μM morphine) and also by the β-adrenoreceptor blocker propranolol (0.1 μM). The α₂-adrenoreceptor antagonist yohimbine (0.1 μM) had no effect on the 5-HT-induced potentiation.
• 3 In the guinea-pig isolated atria the responses to cardioaccelerans nerve stimulation were diminished by 5-HT (0.1 to 1.0 μM). The shift to the right in the frequency-response curve induced by 5-HT (1.0 μM) was additive to the antagonism caused in the atria by propranolol (0.1 μM). The inhibitory effect of 5-HT on the pacemaker responses to nerve stimulation was prevented by the 5-HT receptor antagonists methysergide (1.0 and pizotifen (1.0 μM) and also by the α-adrenoreceptor antagonist phentolamine (0.1 μM).
• 4 The selective α₂-adrenoreceptor agonist clonidine (0.01 μM) reduced to the same extent as 5-HT (1.0 μM) the responses of the guinea-pig atria to nerve stimulation. The inhibitory effect of clonidine was prevented by the α-adrenoreceptor blocker phentolamine (0.1 μM) but not by the 5-HT receptor blocker pizotifen (1.0 μM).
• 5 With the exception of propranolol, which in the atria shifted to the right the concentration-response curve to exogenous noradrenaline (NA), neither 5-HT nor the different antagonists employed modified the sensitivity to NA in the tissues studied.
• 6 The present observations show that 5-HT can produce a dual effect on the sympathetic neurotransmission. It is proposed that a modification in the overflow of NA in response to nerve stimulation is caused by 5-HT and results from the interaction of 5-HT with specific receptors located on the sympathetic fibres. These presynaptic 5-HT receptors behave as excitatory (cat nictitating membrane) or inhibitory (guinea-pig atria) depending on the tissue studied.

     

Potentiating effect of methysergide on norepinephrine-induced constriction of the isolated internal carotid artery of the dog

The stainless steel cannula inserting method was used to examine the effects of methysergide on 5-hydroxytryptamine (5-HT)- and norepinephrine-induced vasoconstriction in the isolated internal carotid artery of the dog. 5-HT, at a dose of 0.3 microgram, induced a marked increase in perfusion pressure, usually over 100 mm Hg. On the other hand, norepinephrine produced a relatively small increase in perfusion pressure (20-40 mm Hg) at a large dose of 10 micrograms. Methysergide inhibited 5-HT-induced vasoconstriction. Norepinephrine-induced vasoconstriction was significantly potentiated by treatment with methysergide and blocked by phentolamine. Methysergide also enhanced the vasoconstrictor response to potassium chloride. Thus, it is suggested that the potentiating effect of methysergide on norepinephrine-induced vasoconstriction may partially be due to activation of the inward calcium channel of the internal carotid artery.     

Operational characteristics of the 5-HT1-like receptors mediating external carotid vasoconstriction in vagosympathectomized dogs

It has recently been shown that the external carotid vasoconstrictor response to 5-HT in the dog is primarily mediated by sumatriptan-sensitive 5-HT₁-like receptors; however, the fact that these receptors are not blocked by metergoline, a 5-HT_1D ligand, raises questions about their possible correlation with the 5-HT_1D receptor subtype. Since a number of drugs display high affinity for the 5-HT_1D (GR127935) and 5-HT_1F (e.g. methysergide and oxymetazoline) receptor subtypes, in this study we have used these drugs to determine whether the above vasoconstrictor 5-HT₁-like receptors correlate with the 5-HT_1D and/or 5-HT_1F receptor subtypes.One-minute intracarotid infusions of 5-HT (0.3–30 g/min), sumatriptan (1–30 g/min), oxymetazoline (0.03–3 g/min) and noradrenaline (0.3–3 g/min) resulted in dose-dependent decreases in external carotid blood flow without changes in arterial blood pressure or heart rate. These vasoconstrictor responses remained unaltered after i.v. administration of physiological saline (0.015, 0.05 and 0.15 ml/kg; n = 4) or ritanserin (1 mg/kg; n = 5). In contrast, GR127935 (1, 3 and 10 g/kg, n = 6) potently blocked the responses to 5-HT (unmasking a dose-dependent vasodilator component) and sumatriptan without affecting those to oxymetazoline or noradrenaline. Interestingly, methysergide (10, 30 and 100 g/kg, n = 5) also blocked the vasoconstrictor responses to 5-HT and sumatriptan, but unlike GR127935, did not revert the vasoconstrictor response to 5-HT; the responses to oxymetazoline remained unaffected, but those to noradrenaline were apparently attenuated by the highest dose.Taken together, the above findings suggest that the sumatriptan-sensitive 5-HT₁-like receptors mediating canine external carotid vasoconstriction resemble 5-HT_1D receptors, probably of the 5-HT_1D subtype on the basis of the resistance to blockade by ritanserin. The pharmacological profile of these receptors could be similar (bovine and human cerebral arteries, porcine carotid arteriovenous anastomoses and human coronary arteries) to other putative 5-HT_1D receptors mediating vascular responses.     

Mechanisms involved in the vasodilator action of 5-hydroxytryptamine in the dog femoral arterial circulation in vivo

The possibility that the vasodilator effect of 5-hydroxytryptamine (5-HT) in vivo involves a presynaptic inhibitory effect on sympathetic nerve activity was investigated in the femoral arterial circulation of pentobarbitone-anaesthetized dogs. The vasodilator effect of intraarterial (i.a.) 5-HT was abolished following ganglion blockade with mecamylamine, and remained inhibited after restoration of femoral vascular tone with i.a. ornipressin. These procedures had no effect on the vasodilator response to i.a. acetylcholine. The vasoconstrictor response to i.a. noradrenaline was not inhibited by mecamylamine. These findings suggest that a presynaptic inhibition of sympathetic neurotransmission is responsible for 5-HT-induced vasodilatation in vivo. Pizotifen (0.1-0.4 mg/kg i.v.) inhibited the 5-HT dilator response, but the doses required were too high to be commensurate with an action at 'D' type 5-HT receptors. Ketanserin (0.1-0.4 mg/kg i.v.) specifically inhibited the dilator response to 5-HT; higher doses (1-4 mg/kg i.v.) also inhibited noradrenaline-induced vasoconstriction. Ketanserin, at all doses used, reduced systemic blood pressure and femoral vascular resistance. The effects of the lower doses of ketanserin (0.1-0.4 mg/kg) cannot be due to peripheral alpha-adrenoceptor antagonism; blockade of the dilator effect of 5-HT may simply be due to inhibition of sympathetic nerve activity by ketanserin itself.     

A pre-junctional action of 5-hydroxytryptamine and methysergide on noradrenergic nerves in dog isolated saphenous vein

Electrical stimulation (2 Hz for 2 min) of dog isolated saphenous vein strips pre-incubated with tritiated noradrenaline increased the overflow of tritium of which about 80% was noradrenaline. 5-Hydroxytryptamine (5-HT; 1·0 × 10^?9-1·0 × 10^?7 mol litre^?1) and methysergide (3·0 × 10^?8-3·0 × 10^?8 mol litre^?1) inhibited the induced overflow of total tritium by a maximum of 78 ± 4% and 47 ± 7% respectively (mean ± s.e. mean, n = 6 for each). Methysergide was about 30 times less potent than 5-HT and the maximum inhibition obtained was less than with 5-HT. Both compounds inhibited electrically-induced contractions and overflow of tritiated noradrenaline. Their inhibitory actions on tritium overflow were little affected by phentolamine (1·0 × 10^?8 mol litre^?1) or cyproheptadine (1·0 × 10^?8 mol litre^?1), nor was the inhibitory effect of methysergide on electrically induced contractions antagonized by atropine, mepyramine, cimetidine or propranolol. The findings suggest that the prejunctional inhibitory effect of methysergide may be mediated via stimulation of a 5-HT receptor which, unlike the D-receptor, is not blocked by cyproheptadine. The possibility that the pre-junctional 5-HT receptor in the dog saphenous vein is the same as the post-junctional receptor in this preparation is discussed.     

AN INVESTIGATION INTO THE MECHANISMS OF THE CARDIOVASCULAR EFFECTS OF 5-HYDROXYTRYPTAMINE IN CONSCIOUS NORMOTENSIVE AND DOCA-SALT HYPERTENSIVE RATS

• 1 The actions of intravenously administered 5-hydroxytryptamine (5-HT) have been analysed in conscious DOCA-salt hypertensive rats using selective 5-HT receptor agonists and antagonists to determine the receptor mechanisms involved and to compare them with those in conscious normotensive rats.
• 2 In both normotensive and hypertensive rats 5-HT, 3 and 10 μg i.v., produced a complex triphasic effect on blood pressure consisting of an initial short lasting depressor response, which was followed by a pressor response and then, finally, a hypotensive phase. Marked decreases in heart rate were observed immediately after dosing, which were followed by small increases in rate.
• 3 The selective 5-HT₃-receptor agonist, 2-methyl 5-HT, 3–30 μg i.v., produced immediate and marked dose-related decreases in blood pressure and heart rate in both normotensive and DOCA-salt hypertensive rats. The 5-HT₃-receptor antagonist, MDL 72222, 0.03 and 0.1 mg/kg i.v., antagonised these effects in both normotensive and DOCA-salt hypertensive rats. Treatment with MDL 72222, 0.3 mg/kg i.v., abolished the initial depressor response and bradycardia produced by 5-HT.
• 4 The 5-HT₂ receptor agonist, α-methyl 5-HT, 3–30 μg i.v., produced dose-related increases in blood pressure which were significantly greater in magnitude in DOCA-salt hypertensive than normotensive rats. Bradycardia was observed consistently at 30 μg only. The 5-HT₂ receptor antagonist, ketanserin, 0.03-0.3 mg/kg i.v., caused a dose-dependent antagonism of the pressor responses produced by α-methyl 5-HT, but had no effect on the increases in blood pressure produced by angiotensin. Ketanserin also antagonised the pressor responses produced by 5-HT in rats pretreated with MDL 72222.
• 5 5-Carboxamidotryptamine (5-CT), the selective ‘5-HT₁-like’ receptor agonist, at doses of 0.1–3 μg i.v. produced dose-related decreases in blood pressure which were more pronounced in the DOCA-salt hypertensive rats than in normotensive rats. These depressor responses were dose-dependently antagonised by methiothepin, 0.3 and 1 mg/kg, i.v. which did not antagonise the depressor responses produced by isoprenaline 0.1 μg/kg i.v. In rats pretreated with MDL 72222 and ketanserin, 5-HT produced dose-related depressor responses which were also antagonised by methiothepin 1 mg/kg i.v. and presumably mediated by 5-HT₁-like receptors.
• 6 In this study, selective 5-HT receptor agonists and antagonists have been used to mimic and block, respectively, the various phases of the 5-HT response. It is concluded that in the conscious rat, the complex cardiovascular effects of 5-HT involve stimulation of at least three different 5-HT receptors (for nomenclature see Bradley, Engel, Feniuk, Fozard, Humphrey, Middlemiss, Mylecharane, Richardson & Saxena, 1986). The initial depressor response and bradycardia involves activation of 5-HT₃-receptors, the secondary vasopressor effect, which is significandy greater in DOCA-salt than normotensive rats results from stimulation of 5-HT₂ receptors and the late vasodepressor response is due to vasodilatation via ‘5-HT₁-like’ receptors.

     

A pre-junctional action of 5-hydroxytryptamine and methysergide on noradrenergic nerves in dog isolated saphenous vein

Electrical stimulation (2 Hz for 2 min) of dog isolated saphenous vein strips pre-incubated with tritiated noradrenaline increased the overflow of tritium of which about 80% was noradrenaline. 5-Hydroxytryptamine (5-HT; 1.0 x 10-9-1.0 x 10-7 mol litre-1) and methysergide (3.0 x 10-8-3.0 x 10-6 mol litre-1) inhibited the induced overflow of total tritium by a maximum of 78 +/- 4% and 47 +/- 7% respectively (mean +/- s.e. mean, n=6 for each). Methysergide was about 30 times less potent than 5-HT and the maximum inhibition obtained was less than with 5-HT. Both compounds inhibited electrically-induced contractions and overflow of tritiated noradrenaline. Their inhibitory actions on tritium overflow were little affected by phentolamine (1.0 x 10-6 mol litre-1) or cyproheptadine (1.0 x 10-6 mol litre-1), nor was the inhibitory effect of methysergide on electrically induced contractions antagonized by atropine, mepyramine, cimetidine or propranolol. The findings suggest that the prejunctional inhibitory effect of methysergide may be mediated via stimulation of a 5-HT receptor which, unlike the D-receptor, is not blocked by cyproheptadine. The possibility that the pre-junctional 5-HT receptor in the dog saphenous vein is the same as the post-junctional receptor in this preparation is discussed.     

INVOLVEMENT OF NITRIC OXIDE IN CORONARY VASCULAR RESPONSES TO 5-HYDROXYTRYPTAMINE IN THE ANAESTHETIZED GREYHOUND

1. The effect of the intracoronary (i.c.) injection of 5-hydroxytryptamine (5-HT; 0.1–1.0 μFg/ kg) was examined before and after inhibition of nitric oxide (NO) synthesis with N-nitro-l-arginine (NOLA; 5 mg/ kg i.c.) in nine anaesthetized greyhounds. Before administration of NOLA, 5-HT increased coronary blood flow (CBF) but decreased large coronary artery diameter indicating simultaneous dilatation of resistance vessels and constriction of large arteries. 2. The administration of NOLA significantly decreased large coronary artery diameter and increased systemic arterial pressure. There was no significant effect on coronary vascular resistance or heart rate. In the presence of NOLA, the 5-HT-induced constriction of the large coronary artery was enhanced and the dilatation of the resistance vessels was reduced. In addition there was a secondary reduction in CBF, a response that was not observed before NOLA treatment. 3. The response to NOLA suggests that a basal release of NO is important in the regulation of coronary and systemic vascular tone. Nitric oxide is an important mediator of coronary vasodilator responses to 5-HT, and in addition the release of NO modulates 5HT-induced constriction of large coronary arteries.     

Allosteric properties of the 5-HT2 receptor system of the rat tail artery

Summary We present an analysis of the interactions of 5-hydroxytryptamine (5-HT) and antagonists (methysergide, ketanserin, ritanserin) with the 5-HT₂ receptor system of strips of rat tail artery. The mode of action of ritanserin was also studied on strips of calf coronary arteries. 1. Ketanserin competitively antagonized 5-HT-induced effects in rat tail artery with an affinity (pK_B = 9.4 nmol/l) consistent with the assumption of an interaction of 5-HT and ketanserin at 5-HT₂-receptors. 2. Methysergide reduced to 50–60% the maximum response to 5-HT in rat tail artery. Concentration-effect curves for 5-HT became biphasic in the presence of methysergide with quickly and slowly developing contractions at low and high concentrations of 5-HT, respectively. 100 nmol/l ketanserin completely restored effects of 5-HT depressed by low concentrations of methysergide (< 10 nmol/l). Higher concentrations of methysergide in the presence of 100 nmol/l ketanserin again depressed the effects of 5-HT. 3. Ritanserin resembles methysergide by causing insurmountable antagonism of 5-HT-induced contractions which can be prevented by ketanserin in both rat tail artery and calf coronary artery. These results are inconsistent with competition between ritanserin and 5-HT for the 5-HT₂ receptor. 4. The findings are consistent with the assumption of an interaction of ketanserin and methysergide or ritanserin with an allosteric site near the 5-HT₂-receptor. Both methysergide and ritanserin appear to antagonize the effects of 5-HT through an allosteric site which is distinct from the 5-HT₂ receptor. Send offprint requests to A. J. Kaumann at the above address     

Functional evaluation of 5-hydroxytryptamine receptor activity in rat resistance vessels

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Effects of methysergide on some cardiovascular responses in the anaesthetized rabbit

Methysergide (0.1?5 mg/kg i.v/) caused dose-dependent reductions in the resting heart rate of anaesthetized rabbits. Resting mean arterial blood pressure was reduced only after the maximum dose. Methysergide 5 mg/kg significantly enhanced the pressor responses to i.v. phenylephrine and enhanced the accompanying bradycadia disproportionately. There was no evidence for a central increase in baroreflex gain, so an action of methysergide on baroreceptor sensory ending is suggested as a possible explanation.     

INHIBITION OF VASODILATATION BY METHYLENE BLUE IN LARGE AND SMALL ARTERIES OF THE DOG HINDLIMB IN VIVO

1. Injection of acetylcholine (ACh, 0.0005-2 micrograms/kg) or glyceryl trinitrate (GTN, 0.01-20 micrograms/kg) into the femoral artery increased femoral artery diameter, femoral blood flow and heart rate, and reduced femoral vascular resistance and systemic arterial blood pressure in anaesthetized dogs. The intravenous (i.v.) injection of ACh (2 micrograms/kg) produced a small decrease in systemic arterial pressure and an increase in heart rate, but did not dilate the hindlimb vessels. 2. Methylene blue, a guanylate cyclase inhibitor, continuously infused into the femoral artery (10 mg/min), attenuated the increase in femoral artery diameter and femoral blood flow, and the decrease in femoral vascular resistance produced by intra-arterial injections of both ACh and GTN. 3. In addition, methylene blue potentiated the decrease in systemic arterial pressure produced by ACh (injected directly into the femoral artery or i.v.), but did not affect the depressor response to GTN. This selective potentiation of ACh-induced hypotension was not affected by autonomic ganglion blockade with hexamethonium (25 mg/kg, i.v.). 4. These results suggest that both ACh- and GTN-induced vasodilatation in vivo occurs through a mechanism involving guanylate cyclase activation in large arteries and resistance vessels in the dog hindlimb. Methylene blue inhibited the local vasodilator actions of ACh in the femoral vasculature despite potentiating the systemic depressor response to that agent.     

Evidence for two types of excitatory receptor for 5-hydroxytryptamine in dog isolated vasculature

1 As part of an investigation into the mode of action of anti-migraine drugs, a study of the excitatory receptors for 5-hydroxytryptamine (5-HT) has been carried out in a range of isolated vascular preparations from the dog.

2 5-HT contracted the dog isolated femoral artery and saphenous vein over the concentration-range 1.0 × 10^-8 to 5.0 × 10^-6 mol/l.

3 In the femoral artery methysergide and cyproheptadine were potent, competitive and specific antagonists of the contractile responses to 5-HT, with pA₂ values of 8.52 and 8.55 respectively.

4 In the saphenous vein, methysergide was only a weak antagonist of 5-HT. In addition, it was an agonist over the concentration-range 5.0 × 10^-8 to 1.0 × 10^-5 mol/l. Cyproheptadine was a weak and unsurmountable antagonist of contractile responses to 5-HT and methysergide.

5 Contractile responses to 5-HT and methysergide in the saphenous vein were not antagonized by morphine (3.0 × 10^-5 mol/l), indomethacin (5.0 × 10^-5 mol/l), phentolamine (5.0 × 10^-7 mol/l), propranolol (1.0 × 10^-6 mol/l), atropine (1.0 × 10^-6 mol/l), mepyramine (1.0 × 10^-6 mol/l) or cimetidine (1.0 × 10^-5 mol/l).

6 In the external carotid and lingual arteries the pattern of activity obtained with methysergide and cyproheptadine was the same as that in the femoral artery, while in the auricular artery the pattern of activity was the same as that in the saphenous vein.

7 The results are consistent with the hypothesis that there are two types of receptor mediating 5-HT-induced vasoconstriction in dog vasculature. One type, characterized by the pattern of activity obtained in the femoral artery, is like the previously described `D-receptor'. The other type, characterized by the pattern of activity obtained in the saphenous vein, has not been described before. The verification of this hypothesis requires the identification of a specific antagonist of 5-HT and methysergide in the saphenous vein.

     

The selective carotid arterial vasoconstrictor action of GR43175 in anaesthetized dogs

1. GR43175 is a highly selective agonist at 5-HT1-like receptors in the dog saphenous vein. This study describes the haemodynamic effects of GR43175 in barbitone-anaesthetized dogs. 2. GR43175 (1-1000 micrograms kg-1, i.v.) produced dose-dependent decreases in carotid arterial blood flow with little or no change in arterial blood pressure. The decrease in blood flow was associated with an increase in carotid arterial vascular resistance. In preliminary studies, the dose of GR43175 producing 50% of the maximum carotid vasoconstrictor response was 39 +/- 8 micrograms kg-1, i.v. 3. In comparative regional haemodynamic studies, GR43175 (1-1000 micrograms kg-1, i.v.) had little effect on total peripheral resistance or resistance in the mesenteric, vertebral and coronary arterial vascular beds. Low doses of GR43175 decreased, whilst high doses (100 micrograms kg-1, i.v. and above) increased femoral arterial vascular resistance. GR43175 (1-1000 micrograms kg-1, i.v.) had no effect on respiratory inflation pressure. In doses of 100 micrograms kg-1 i.v. and above, GR43175 caused small decreases in heart rate. 4. The carotid arterial vasoconstrictor action of GR43175 was resistant to antagonism by the 5-HT2 receptor, 5-HT3 receptor and alpha-adrenoceptor blocking drugs, ketanserin, MDL72222 and phentolamine respectively, but could be antagonized by the non-selective 5-HT1-like receptor blocking drug methiothepin. Methiothepin had no effect on the carotid vasoconstrictor action of the thromboxane A2 mimetic, U46619. 5. The results demonstrate that GR43175 produces a selective vasoconstriction in the carotid arterial circulation of anaesthetized dogs via activation of 5-HT1-like receptors, which appear similar to those mediating contraction of the dog isolated saphenous vein.     

Differential effects of glibenclamide on responses to thromboxane A2 mimic, U46619, in the pulmonary and hindquarters vascular beds of the cat

The inhibitory effects of the oral sulfonylurea, glibenclamide, on vasoconstrictor responses to the thromboxane A₂ mimic, U46619, were investigated in the pulmonary and hindquarters vascular beds of the cat under constant flow conditions. When lobar arterial tone was at resting conditions (14±2 mm Hg), intralobar injections of U46619, prostaglandin F_2α, prostaglandin D₂, angiotensin II, norepinephrine, and BAY K 8644 caused dose-related increases in lobar arterial pressure without altering left atrial pressure. Following an intralobar infusion of glibenclamide (5 mg/kg), vasoconstrictor responses to U46619, prostaglandin F_2α and prostaglandin D₂ were significantly reduced, whereas vasoconstrictor responses to norepinephrine and angiotensin II were not altered and responses to BAY K 8644 were significantly enhanced. When tone in the pulmonary vascular bed was raised to a high steady level (36±3 mm Hg), glibenclamide in a dose of 5 mg/kg i.a. markedly attenuated responses to injections of U46619 and reduced the vasodilator responses to the K⁺-ATP channel opener, levcromakalim, whereas responses to acetylcholine and S-nitroso-N-acetylpenicillamine (SNAP), a nitric oxide donor, were not changed. In the hindquarters vascular bed of the cat, administration of glibenclamide in a dose of 5 mg/kg i.a. had no significant effect on vasoconstrictor responses to U46619, norepinephrine or angiotensin II. Hindquarters vasodilator responses to levcromakalim, but not to nitric oxide, were decreased significantly following administration of glibenclamide. These data suggest that glibenclamide, in addition to inhibiting K⁺-ATP channels, has thromboxane A₂ receptor blocking activity in the pulmonary vascular bed of the cat. These data also suggest that vasoconstrictor responses to U46619 may be mediated by different thromboxane A₂ receptors with different binding affinities in the pulmonary and in the hindquarters vascular beds of the cat.