Lack of interaction between alpha(2)-autoreceptors and prejunctional receptors mediating a facilitatory effect on noradrenaline release.

The present study was undertaken to investigate the effect of alpha(2)-autoreceptor blockade on the facilitatory influence exerted by activation of beta -, A(2A)-adenosine- and angiotensin II receptors. Segments of a rat-tail artery, previously incubated with(3)H-noradrenaline, were subjected to electrical stimulation. The influence of isoprenaline, the compound CGS21680 and angiotensin II on the overflow of tritium evoked by electrical stimulation was checked before and after alpha(2)-adrenoceptor blockade. All the agonists used caused concentration-dependent increases of tritium overflow, the maximal effect representing increases of 44.2, 27.4 and 41.2% for isoprenaline, CGS21680 and angiotensin II, respectively. In the presence of alpha(2)-adrenoceptor blockade by phenoxybenzamine ( 1 microm) or yohimbine (33 or 100 nm), the facilitatory influence of isoprenaline, CGS21680 and angiotensin II was not significantly changed. Since this facilitatory influence, which involves the activation of G(s)- or G(q)-proteins, was not enhanced by alpha(2)-adrenoceptor blockade, it is concluded that the enhancement of the negative modulation resulting from activation of A(1)-adenosine-, muscarine- and kappa -receptors, as previously shown, should be due to the fact that the involved systems share signal transduction mechanisms, or at least G-proteins.

A comparative study of postsynaptic alpha2-adrenoceptors of the dog mesenteric and rat femoral veins.

The aim of the present study was to compare the subtype of postjunctional alpha2-adrenoceptors of canine mesenteric and rat femoral veins. To check whether the presence of alpha1-adrenoceptors in both tissues might interfere with alpha2-adrenoceptor-mediated effects, the experiments were carried out under two experimental conditions: without and with alpha1-adrenoceptor blockade. The selective and irreversible alpha1-adrenoceptor antagonist phenoxybenzamine (30 nM) was used to eliminate alpha1-adrenoceptors. The pA2 values for the antagonism exerted by eight alpha-adrenoceptor antagonists (rauwolscine, yohimbine, RX821002, WB4101, idazoxan, phentolamine, spiroxatrine and prazosin) against the highly selective alpha2-adrenoceptor agonist UK-14,304 were determined under these two experimental conditions and correlated with pKi values of the same antagonists at cloned human alpha2A-, alpha2B- and alpha2C-adrenoceptors expressed in Chinese hamster lung cells and at the alpha2D-adrenoceptors in the rat submaxillary gland or the bovine pineal gland. In most of the experiments carried out in the canine mesenteric vein, the concentration-response curves for UK-14,304 were biphasic; the first phase was antagonized by low concentrations (2-50 nM) of the antagonists used except prazosin, while the second one was antagonized by 30 nM of either prazosin or phenoxybenzamine. In the rat femoral vein, the concentration-response curves to UK-14,304 were monophasic. In either tissue, the pA2 values obtained in untreated preparations and in preparations pretreated with phenoxybenzamine were not significantly different, showing that effects resulting from the activation of alpha1-adrenoceptors can be avoided simply by using low concentrations of the highly selective alpha2-adrenoceptor agonist UK-14,304. The correlation between pA2 and pKi values showed that, while in the canine mesenteric vein the postjunctional alpha2-adrenoceptors resemble alpha2A-adrenoceptors more closely, in the rat femoral vein they are more closely related to alpha2D-adrenoceptors. In either species, therefore, they belong to the genetic alpha2A/D type of alpha2-adrenoceptor.

Prejunctional effects of angiotensin II and bradykinin in the heart and blood vessels.

1. Angiotensin and bradykinin facilitate the release of noradrenaline from sympathetic nerve terminals and cause positive inotropy in rat isolated atria and ventricles. The effect of bradykinin was enhanced by the ACE inhibitor, ramiprilat. 2. The facilitated release of noradrenaline in rat ventricle by bradykinin was blocked by the beta2-receptor antagonist HOE-140. This response is also reduced by removing the endocardium, suggesting the release of a mediator from the endocardium. 3. The facilitated noradrenaline release by angiotensin II and bradykinin was blocked by the angiotensin receptor antagonist saralasin to the same extent. In contrast, losartan caused only minor blockade in a range of vascular and cardiac tissues. This suggests that angiotensin and bradykinin exert these responses by interacting with a prejunctional receptor different from the established AT1 subtype. 4. These results suggest that bradykinin mediates facilitation of noradrenaline release via the local release of angiotensin onto an atypical AT1 receptor.

Prejunctional alpha2A-autoreceptors in the human gastric and ileocolic arteries.

This study was undertaken to determine the subtype of prejunctional alpha2-autoreceptors in human blood vessels. Segments of gastric and ileocolic arteries were incubated with [3H]noradrenaline and subsequently perifused with modified Krebs-Henseleit solution containing cocaine (12 microM). Five periods of electrical stimulation (S1-S5) were applied (1 Hz, 1 ms, 50 V for 1 min). Concentration-response curves for the facilitatory effect of eight alpha-adrenoceptor antagonists [rauwolscine, 2-[2-(2-methoxy-1,4-benzodioxanyl)] imidazoline (RX821002), yohimbine, phentolamine, idazoxan, 2-(2',6'-dimethoxyphenoxyethyl)-aminomethyl-1,4-benzodioxan (WB4101), spiroxatrine and prazosin] were determined. All antagonists enhanced the stimulation-evoked overflow of tritium, indicating the existence of alpha2-autoreceptors. The EC30% values of the antagonists (concentrations that increased the evoked overflow of tritium by 30%) were taken as a measure of affinity to the autoreceptors. Correlations between the pEC30% values obtained in the present study and the pKi values of the same antagonists at cloned human alpha2A-, alpha2B-, alpha2C-adrenoceptors expressed in Chinese hamster lung cells and at alpha2D-adrenoceptors in the rat submaxillary gland or the bovine pineal gland showed that the alpha2-autoreceptors in the human gastric and ileocolic arteries resemble most closely the alpha2A-subtype.

The role of MAO-A and MAO-B in the metabolic degradation of noradrenaline in human arteries.

1. Segments of human cystic, gastric and ileocolic arteries were obtained from patients undergoing surgery. 2. Segments of arterial tissues, the noradrenaline content of which ranged between 0.27 and 0.52 microg g(-1), were incubated with 0.1 micromol l(-1) [3H]-noradrenaline for 30 min and the accumulation of the amine as well as the formation of metabolites was measured. 3. In all the arteries, oxidative deamination predominated over O-methylation; the mean values of the deaminated and O-methylated metabolites formed for the three arteries were 247.6 and 82.4 pmol g(-1) tissue, respectively. Dihydroxymandelic acid (DOMA) was the most abundant metabolite. 4. Both clorgyline (a selective MAO-A inhibitor) and selegiline (a selective MAO-B inhibitor) reduced the formation of dihydroxyphenylglycol (DOPEG), DOMA and O-methylated-deaminated metabolites (OMDA), and increased that of normetanephrine (NMN). However, clorgyline depressed the formation of DOPEG more than that of DOMA, while selegiline depressed the formation of DOMA more than that of DOPEG. 5. In conclusion, three major differences distinguish the metabolism of noradrenaline by human arteries from that observed in other species: (1) the large predominance of deamination over O-methylation; (2) the extremely high formation of DOMA; and (3) the relative lack of selectivity of clorgyline and selegiline for MAO-A and B, respectively. Since the arterial vessels used were collected from patients older than 60 years, the morphological changes depending on age may explain the increase in DOMA formation.

Different receptors for angiotensin II at pre- and postjunctional level of the canine mesenteric and pulmonary arteries.

1. This investigation was undertaken to compare pre- and postjunctional receptors involved in the responses of the canine mesenteric and pulmonary arteries to angiotensin II. 2. In the mesenteric artery, angiotensin II caused an enhancement of tritium overflow evoked by electrical stimulation (EC30% = 5 nM), the maximal effect representing an increase by about 45%. Postjunctionally, angiotensin II caused concentration-dependent contractions (pD2 = 8.57). Saralasin antagonized both pre- and postjunctional effects of angiotensin II, but it was more potent at post- than at prejunctional level (pA2 of 9.51 and 8.15, respectively), while losartan antagonized exclusively the postjunctional effects of angiotensin II (pA = 8.15). PD123319 had no antagonist effect either pre- or postjunctionally. 3. In the pulmonary artery, angiotensin II also caused an enhancement of the electrically-evoked tritium overflow (EC30% = 1.54 nM), its maximal effect increasing tritium overflow by about 80%. Postjunctionally, angiotensin II caused contractile responses (pD2 = 8.52). As in the mesenteric artery, saralasin antagonized angiotensin II effects at both pre- and postjunctional level and it was more potent postjunctionally (pA2 of 9.58 and 8.10, respectively). Losartan antagonized only the postjunctional effects of angiotensin II (pA2 = 7.96) and PD123319 was ineffective. 4. It is concluded that in both vessels: (1) pre- and postjunctional receptors belong to a different subtype, since they are differently antagonized by the same antagonists; (2) postjunctional receptors belong to AT1 subtype, since they are blocked by losartan but not by AT2 antagonists; (3) prejunctional receptors apparently belong to neither AT1 or AT2 subtype since they are blocked by neither AT1 nor AT2 antagonists.

Influence of maturation and ageing on the biotransformation of noradrenaline in the rat.

The present investigation was undertaken to study the influence of maturation and ageing on the disposition of noradrenaline by the aorta, heart (ventricle), liver and kidney of the rat. Slices of these tissues taken from rats aged less than 18 h, 2.5-3 months or 18-24 months were incubated with 0.1 mumol.l-1 3H-amine during 30 min. At the end of this period, the accumulation of the intact amine in the tissue, as well as the 3H-metabolites formed (3,4-dihydroxyphenylethylglycol, 3,4-dihydroxymandelic acid, normetanephrine and O-methylated deaminated metabolites) were determined by scintillation counting. The results obtained show that in the rat: 1) at any age, noradrenaline is preferentially deaminated; 2) while the capacity of the sympathetic nerve terminals in accumulating noradrenaline is rather well developed at birth, the metabolic system for its degradation is still immature; 3) aldehyde dehydrogenase activity or that of its co-factor (or both) of the heart is apparently missing at birth; 4) removal of noradrenaline by the liver and the kidney did not change with ageing, while that by the aorta decreased and that by the heart increased.

Alpha-adrenoceptor-mediated prejunctional effects of chloroethylclonidine in the canine saphenous vein.

The present study was undertaken to look for the effect of chloroethylclonidine (CEC) on prejunctional alpha-2 autoreceptors of the canine saphenous vein. The effect was tested on tritium overflow evoked by electrical stimulation from tissues preloaded with 0.2 microM 3H-norepinephrine. Yohimbine (3-300 nM) and CEC (1-125 microM) increased and UK-14,304 reduced the overflow of tritium evoked by 300 pulses (1 Hz). The maximal increase of tritium overflow caused by yohimbine was much higher than that caused by CEC: 3.82 and 1.74 times, respectively. CEC (5 microM) abolished both the inhibition caused by UK-14,304 and the enhancement of tritium overflow caused by yohimbine. However, when CEC was added after yohimbine, it reduced the electrically evoked overflow of tritium, the maximal effect being a reduction of tritium overflow by 35%. Prazosin (1-100 nM) did not change either the inhibitory effect of UK-14,304 or the facilitatory effect of CEC. These results suggest that CEC acts on two different subtypes of prejunctional alpha-2 autoreceptors; on one of them it acts as an antagonist and increases the electrically evoked overflow of tritium (and inhibits both the effect of UK-14,304 and yohimbine); on the other it acts as an agonist and reduces the electrically evoked overflow of tritium. Alternatively, one can admit that CEC is able to inhibit alpha-2 autoreceptors, which causes an increase of the transmitter release, and to activate a nonadrenergic inhibitory receptor thus causing a reduction of the transmitter release.

Prejunctional alpha2A-autoreceptors in the canine saphenous vein.

The aim of the study was to determine the subtype of prejunctional alpha2-autoreceptors in the canine saphenous vein. Segments of the vein were incubated with 3H-noradrenaline and subsequently perifused with modified Krebs-Henseleit solution. Five periods of electrical stimulation were applied (S1-S5; each for 2 min, 1 Hz). Concentration-response curves for the inhibitory effect of the alpha2-adrenoceptor agonists oxymetazoline and UK-14,304 and for the facilitatory effect of nine antagonists were determined. Correlations between the pEC30%s for the antagonists obtained in the present study and the pKis for the same antagonists obtained in tissues expressing only alpha2A- (HT29 cells), alpha2B- (neonatal rat lung), alpha2C- (OK cells) or alpha2D-adrenoceptors (rat submaxillary gland) showed that the alpha2-autoreceptors in the canine saphenous vein resemble most closely the alpha2A-subtype. Furthermore, oxymetazoline was a highly potent agonist (pIC50% = 8.10) and prazosin was a weak antagonist (pEC30% = 6.46), confirming that the alpha2-adrenoceptors involved in the modulation of the response to electrical stimulation of the canine saphenous vein do not belong to either the alpha2B- or alpha2C-subtype. On the other hand, the EC30% ratios phentolamine/rauwolscine and idazoxan/rauwolscine were much closer to the ratios obtained at alpha2-autoreceptors of the rabbit- (alpha2A) than of the guinea-pig brain cortex (alpha2D). The results suggest that the sympathetic nerves of the canine saphenous vein are endowed with alpha2A-adrenoceptors.

Endothelium-dependent vascular responses in 1,3-dipropyl-8-sulphophenylxanthine (DPSPX) hypertensive rats.

The study was undertaken to test the endothelium-mediated vascular responses in rats rendered hypertensive by chronic administration of 1,3-dipropyl-8-sulphophenylxanthine (DPSPX). The relaxant effect of carbachol (an endothelium-dependent relaxing drug) and of sodium nitroprusside (an endothelium-independent relaxing drug) as well as the potentiation of the contractile effect of noradrenaline by NG-nitro-L-arginine methyl ester (L-NAME) were compared in aortic rings from normotensive and DPSPX-hypertensive rats. Carbachol and sodium nitroprusside caused concentration-dependent relaxations in aortic rings precontracted by 1 microM noradrenaline. The relaxant effect being of carbachol was significantly reduced in tissues of DPSPX-hypertensive rats: the maximal relaxant effect being 86 +/- 3% and 64 +/- 4% (of the pre-existing tone) in normal and hypertensive rats, respectively, while there were no significant differences in the relaxant effect of sodium nitroprusside. L-NAME (100 microM) significantly reduced the EC50 values of noradrenaline (3.71 +/- 0.28 times, n = 8 and 2.96 +/- 0.27 times, n = 7, in normal and hypertensive rats, respectively) and significantly enhanced the maximal contractile effect of noradrenaline (46 +/- 8%, n = 8 and 35 +/- 6%, n = 7, in normal and hypertensive rats respectively): the factors of reduction of EC50 values and the percentages of enhancement of the maximal contractile effect in the aorta of normal and hypertensive rats were not significantly different. The results obtained provide evidence of functional impairment of the endothelium in DPSPX-hypertensive rats.

Homogeneous or heterogeneous distribution of systemically administered adrenaline: organ dependence.

In incubation experiments it was shown that exogenous adrenaline or noradrenaline does not distribute homogeneously into the adrenergic varicosities of the rat vas deferens (wall with thick and compact muscle layer) but does distribute homogeneously in the rat spleen capsule (thin and loose muscle layer, containing more extracellular space than the vas deferens). To circumvent any hypothetical role of the muscular layer in the distribution of the amine, 100 micrograms.kg-1.h-1 adrenaline was administered to rats in vivo either i.v. (during 90 min) or i.p. (under pentobarbital anaesthesia, an Alzet minipump was implanted which delivered that dose during 6 days). The rats also received 100 mg.kg-1 pargyline (to inhibit MAO) and 100 mg.kg-1 tropolone (to inhibit COMT). At the end of adrenaline administration, vasa deferentia and spleen capsule were removed, washed and then exposed to 100 mumol.l-1 tyramine for 20 min. At the end of this exposure, the ratio noradrenaline/adrenaline in the tissue and in the medium was compared. In the vas deferens both after i.v. and i.p. administration of adrenaline, the ratio noradrenaline/adrenaline was about 3 times higher in the medium than in the tissue, while in the spleen capsule the ratio noradrenaline/adrenaline was not significantly different in the medium and in the tissue. We conclude that, even when the amine reaches the storage sites from the blood, it distributes homogeneously in the spleen capsule and heterogeneously in the vas deferens, perhaps because there are more than one kind of storage vesicles in the vas deferens.

Pre- and postjunctional effects of 5-hydroxytryptamine in dog cephalic veins of neonates.

1. This investigation was undertaken to determine the existence of pre- and postjunctional effects of 5-hydroxytryptamine (5-HT) in the cephalic vein of newborn dogs. For the sake of comparison, some experiments were also carried out on the veins of adult animals. 2. 5-HT reduced, in a concentration-dependent manner the overflow of tritium evoked by electrical stimulation of tissues previously loaded with 3H-noradrenaline for the same range of concentrations (0.01-1 microM 5-HT) in both neonates and adults. The maximal reduction of tritium overflow caused by 5-HT was 36 +/- 6% and 77 +/- 5% (n = 4; P < 0.001) in neonates and adults, respectively. 3. Postjunctionally, 5-HT caused concentration-dependent contractions of vessels from both neonates and adults and its EC50 values were not significantly different at the two ages. The maximal effect of 5-HT was smaller in newborn (0.69 +/- 0.03 g/mg) than in adult (1.16 +/- 0.06 g/mg) animals (n = 9; P < 0.01). 4. Metitepin antagonized 5-HT effects while ketanserin was ineffective, both pre- and postjunctionally, in newborn and adult dogs. 5. It is concluded that there are 5-HT receptors at pre- and postjunctional level at birth and that these receptors probably belong to the 5-HT1 type.

Hypertension and enhanced beta-adrenoceptor-mediated facilitation of noradrenaline release produced by chronic blockade of adenosine receptors.

1. The study was undertaken to compare the beta-adrenoceptor-mediated facilitation of noradrenaline release in the tail artery of vehicle-treated rats and of rats rendered hypertensive by chronic administration of 1,3-dipropyl-8-sulphophenylxanthine (DPSPX). Artery rings were loaded with [3H]-noradrenaline, and five periods of electrical stimulation (1 Hz for 2 min) were applied. To eliminate the influence of prejunctional alpha 2-adrenoceptors, the tissues were pre-exposed to 1 microM phenoxybenzamine. 2. Isoprenaline caused a concentration-dependent increase of tritium overflow elicited by electrical stimulation. It was more effective in arteries from DPSPX-treated than in those from vehicle-treated rats; isoprenaline (27.8 nM) increased by 30% tritium overflow in vessels from vehicle-treated rats whereas isoprenaline (7.0 nM) produced a 30% increase in vessels from DPSPX-treated animals. Furthermore, the maximal effect of isoprenaline was a 32.6% increase in control rats but a 48.6% increase in DPSPX-treated rats. 3. These results show that the sympathetic nerve endings of the rat tail artery are endowed with prejunctional beta-adrenoceptors which mediate facilitation of noradrenaline release elicited by electrical stimulation. They also suggest that adenosine receptors and beta-adrenoceptors interact at the prejunctional level and that impairment of this 'talk' may lead to the development of a hypertensive state.

Long-term administration of 1,3-dipropyl-8-sulphophenylxanthine (DPSPX) alters alpha 2-adrenoceptor-mediated effects at the pre- but not at the postjunctional level.

The present investigation was undertaken to see whether a long-term inhibition of adenosine receptors--leading to hypertension--interferes with alpha 2-adrenoceptor-mediated modulation of noradrenaline release. Rat tail arteries were removed from normal and from hypertensive animals obtained by chronic treatment with intraperitoneally infused DPSPX (1,3-dipropyl-8-sulphophenylxanthine) or orally administered L-NAME (NG-Nitro-L-arginine methyl ester). To study prejunctional effects, the influence of UK-14,304 (5-bromo-6(imidazoline-2-ylamino)-quinoxaline) and yohimbine on the overflow of tritium evoked by electrical stimulation (100 V; 1 Hz; 2 ms; 5 min) from tissues preloaded with 3H-noradrenaline was analysed. To study postjunctional effects, concentration-response curves to UK-14,304 were determined. In DPSPX-treated rats there was an enhancement of the prejunctional effects of UK-14,304: its Ec30% was reduced from 381 (250; 579) to 85 (73; 99) nmol.l-1 (n = 5; P < 0.05) and its maximal effect--expressed as percent reduction of tritium overflow-increased from 45 +/- 5% to 61 +/- 5% (n = 6; P < 0.05). In L-NAME-treated rats there was no change in either of these two parameters. At the postjunctional level, there was no change in the sensitivity to UK-14,304 in tissues from either DPSPX- or L-NAME-treated rats. Yohimbine (10-1000 nmol.l-1) caused a concentration-dependent increase of tritium overflow evoked by electrical stimulation in both control and hypertensive animals (either DPSPX- or L-NAME-treated).(ABSTRACT TRUNCATED AT 250 WORDS)

Postnatal development of vascular beta-adrenoceptor-mediated responses and the increase in the adrenaline content of the adrenal gland have a parallel time course.

The present study was undertaken to analyse the relationship between postnatal development of vascular beta 2-adrenoceptor-mediated responses and the content of adrenaline in the adrenal gland and its concentration in plasma. Dog saphenous vein tissue from newborn, two-weeks old and adult animals were either preloaded with 3H-noradrenaline (or 3H-adrenaline) to study prejunctional beta-adrenoceptor-mediated effects or mounted in organ baths to determine isoprenaline-induced relaxation of preparations contracted by phenylephrine to about 65% of the maximum. The adrenal glands and samples of blood from the same animals were taken for estimation of adrenaline and noradrenaline. At birth, there were no beta-adrenoceptor-mediated effects pre- or postjunctionally. At two weeks, while the results at the prejunctional level were not significantly different from those obtained in newborns, at the postjunctional level there was a relaxant response to isoprenaline, which antagonised about 35% of the previous contraction to 1.75 mumol.l-1 phenylephrine. In adults, isoprenaline (50 nmol.l-1) increased by 24% tritium overflow evoked by electrical stimulation of tissues preloaded with 3H-noradrenaline but not that of tissues preloaded with 3H-adrenaline. On the other hand, propranolol (1 mumol.l-1) reduced by 21% the overflow of tritium evoked by electrical stimulation of tissues preloaded with 3H-adrenaline but not that of tissues preloaded with 3H-noradrenaline; postjunctionally, the maximal response to isoprenaline antagonised 70% of the previous contraction to 1.75 mumol.l-1 phenylephrine.(ABSTRACT TRUNCATED AT 250 WORDS)

Lack of pre- and postjunctional beta adrenoceptor-mediated effects in the canine saphenous vein at birth.

The present study was undertaken to compare the relevance of alpha and beta adrenoceptor-mediated responses at pre- and postjunctional level in the canine saphenous vein of neonates and adults. To quantify prejunctional action, the effect of drugs on the neurogenic outflow of tritium from the vessel loaded previously with [3H]norepinephrine or [3H]epinephrine was measured. The selective alpha-2 adrenoceptor agonist UK-14,304 [5-bromo-6-(2-imidazoline-2-ylamino)-quinoxaline; 10-1000 nM reduced and the selective alpha-2 adrenoceptor antagonist yohimbine (30-300 nM) enhanced the overflow of tritium evoked by electrical stimulation (1 Hz; 2 msec; 100 V; 300 pulses) in both adult and neonate tissues. However, in strips preloaded with [3H]norepinephrine, the beta adrenoceptor agonist isoproterenol (50 nM) increased the overflow of tritium in strips of adults but had no effect in strips of neonates; and in the strips preloaded with [3H]epinephrine, the beta adrenoceptor antagonist propranolol (1 microM) reduced the overflow of tritium in adults but had no effect in neonates. Postjunctionally, phenylephrine (0.1-50 microM) caused concentration-dependent contractions of the saphenous vein rings from adults and neonates but isoproterenol, which caused concentration-dependent relaxations on rings contracted previously by phenylephrine in adults, had no effect in neonates. In contrast to isoproterenol, forskolin (0.05-5 microM), under the same conditions, caused concentration-dependent relaxations of rings of both adults and neonates.(ABSTRACT TRUNCATED AT 250 WORDS)

A comparative study of the distribution of tritiated and endogenous noradrenaline in the rat vas deferens and in the dog spleen capsule.

The aim of the present work was to study the influence of tissue morphological characteristics on the neuronal release (and by inference the distribution) of tritiated and endogenous noradrenaline. Rat vas deferens and dog spleen capsule were loaded with 0.2 mumol/l 3H-noradrenaline, after inhibition of the noradrenaline metabolizing enzymes. Some preparations were washed out under control conditions (spontaneous efflux) and others were washed out in the presence of the releasing agents: 40 mumol/l of Ro 4-1284 (a reserpine-like compound), 100 mmol/l potassium or 100 mumol/l tyramine. The fractional rate of loss (efflux/tissue content) of each amine was determined and the ratio "endogenous amine/3H-noradrenaline" in the efflux and in the tissue were also calculated. The results showed no preferential release of one of the amines in the spleen capsule, whereas a preferential release of tritiated noradrenaline was observed in the vas deferens. The smooth muscle layer in the vas deferens was much thicker and more compact than that of the spleen capsule. The 3H-sorbitol space was smaller in the former than in the latter. We conclude that the morphological characteristics of the tissues contribute to the differences in 3H-noradrenaline distribution in the adrenergic varicosities of these preparations.

Supersensitivity to catecholamines after inhibition of extraneuronal uptake (uptake2) or O-methylation.

The influence of inhibition of uptake2 and of O-methylation on beta-adrenoceptor-mediated responses of guinea-pig trachea and rat uterus to some catecholamines was compared. The responses were more markedly enhanced by inhibition of O-methylation (by U-0521) than by inhibition of uptake2 (by hydrocortisone). The ranking order for the degree of super-sensitivity caused by either U-0521 or hydrocortisone was different in the two tissues, suggesting differences between the O-methylating systems of those tissues. In experiments with 3H-isoprenaline metabolism, hydrocortisone had a weak inhibitory effect on O-methylation in both tissues.

Hypothermia induces supersensitivity of mouse vas deferens to adrenergic agonists: evidence for postjunctional alpha 2-adrenoceptors.

Hypothermia-induced potentiation of alpha-adrenoceptor-mediated responses of the mouse vas deferens to noradrenaline is mainly caused by inhibition of the sites of loss. However, even after blockade of the sites of loss for noradrenaline or when using methoxamine (which is not a substrate for uptake or metabolism) hypothermia still causes a significant increase in responsiveness. This remaining supersensitivity was shown to be an increased receptor affinity. Furthermore, hypothermia revealed contractile responses to low concentrations of UK 14304 (an alpha 2-agonist).

The actions of 5-hydroxytryptamine receptor agonists and antagonists at pre- and postjunctional level on the canine saphenous vein.

The prejunctional and postjunctional 5-HT receptors of the canine saphenous vein were studied. The release of 3H-noradrenaline (3H-NA) from incubated saphenous vein strips was inhibited by 5-hydroxytryptamine (5-HT) in a concentration-dependent way (5-HT concentrations: 0.01, 0.1 and 1.0 mumol.l-1), but not by the selective 5-HT1A agonist 8-hydroxy-dipropylaminotetralin (8-OH-DPAT; 1 and 10 mumol.l-1). The inhibitory effect of 5-HT was antagonized by metitepine and methysergide, but not by yohimbine, (-)-pindolol or ketanserin. In strips preincubated with 5-HT (1.2 mumol.l-1), the fractional release of 3H-NA was slightly reduced (paired experiments). 5-HT and 8-OH-DPAT caused concentration-dependent contractions of the saphenous smooth muscle. A parallel shift of the concentration-response curve for 8-OH-DPAT to the right was caused by metitepine and yohimbine, but not by ketanserin. The contractions caused by 5-HT were antagonized by metitepine and yohimbine (parallel displacement of the curves to the right), as well as by ketanserin and methysergide (with a depression of the upper part of the curve). Blockade of alpha-adrenoceptors (due to prazosin plus a low concentration of yohimbine) also resulted in a weak antagonistic effect. Ketanserin and metitepine displaced the noradrenaline concentration-response curve to the right. We conclude that the saphenous vein of the dog is endowed with prejunctional receptors of the 5-HT1 type which can not be classified as belonging either to the 1A or 1B subtype; and that at the postjunctional level 5-HT1 (possibly of the 1D subtype) and 5-HT2 receptors are present.(ABSTRACT TRUNCATED AT 250 WORDS)