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We explored whether the M2 muscarinic receptor in the guinea pig ileum elicits a highly potent, direct-contractile response, like that from the M3 muscarinic receptor knockout mouse. First, we characterized the irreversible receptor-blocking activity of 4-DAMP mustard in ileum from muscarinic receptor knockout mice to verify its M3 selectivity. Then, we used 4-DAMP mustard to inactivate M3 responses in the guinea pig ileum to attempt to reveal direct, M2 receptor-mediated contractions. The muscarinic agonist, oxotremorine-M, elicited potent contractions in ileum from wild-type, M2 receptor knockout, and M3 receptor knockout mice characterized by negative log EC50 (pEC 50 ) values ± SEM of 6.75 ± 0.03, 6.26 ± 0.05, and 6.99 ± 0.08, respectively. The corresponding E max values in wild-type and M2 receptor knockout mice were approximately the same, but that in the M3 receptor knockout mouse was only 36% of wild type. Following 4-DAMP mustard treatment, the concentration–response curve of oxotremorine-M in wild-type ileum resembled that of the M3 knockout mouse in terms of its pEC 50 , E max, and inhibition by selective muscarinic antagonists. Thus, 4-DAMP mustard treatment appears to inactivate M3 responses selectively and renders the muscarinic contractile behavior of the wild-type ileum similar to that of the M3 knockout mouse. Following 4-DAMP mustard treatment, the contractile response of the guinea pig ileum to oxotremorine-M exhibited low potency and a competitive-antagonism profile consistent with an M3 response. The guinea pig ileum, therefore, lacks a direct, highly potent, M2-contractile component but may have a direct, lower potency M2 component.

Flocalin (FLO) is a new ATP-sensitive K (K) channel opener (KCO) derived from pinacidil (PIN) by adding fluorine group to the drug's structure. FLO acts as a potent cardioprotector against ischemia-reperfusion damage in isolated heart and whole animal models primarily via activating cardiac-specific Kir6.2/SUR2A K channels. Given that FLO also confers relaxation on several types of smooth muscles and can partially inhibit l-type Ca channels, in this study, we asked what is the mechanism of FLO action in bladder detrusor smooth muscle (DSM). The actions of FLO and PIN on contractility of rat and guinea pig DSM strips and membrane currents of isolated DSM cells were compared by tensiometry and patch clamp. Kir6 and SUR subunit expression in rat DSM was assayed by reverse transcription PCR (RT-PCR). In contrast to PIN (10 μM), FLO (10 μM) did not produce glibenclamide-sensitive DSM strips' relaxation and inhibition of spontaneous and electrically evoked contractions. However, FLO, but not PIN, inhibited contractions evoked by high K depolarization. FLO (40 μM) did not change the level of isolated DSM cell's background K current, but suppressed by 20 % l-type Ca current. Determining various Kir6 and SUR messenger RNA (mRNA) expressions in rat DSM by RT-PCR indicated that dominant K channel in rat DSM is of vascular type involving association of Kir6.1 and SUR2B subunits. Myorelaxant effects of FLO in bladder DSM are explained by partial blockade of l-type Ca channel-mediated Ca influx rather than by hyperpolarization associated with increased K permeability. Thus, insertion of fluorine group in PIN's structure made the drug more discriminative between Kir6.2/SUR2A cardiac- and Kir6.1/SUR2B vascular-type K channels and rendered it partial l-type Ca channel-blocking potency. [ABSTRACT FROM AUTHOR]

The A adenosine and M muscarinic receptors exert protective (including energy consumption limiting) effects in the heart. We investigated the influence of adenosine deaminase (ADA) inhibition on a representative energy consumption limiting function, the direct negative inotropic effect elicited by the A adenosinergic and M muscarinergic systems, in eu- and hyperthyroid atria. Furthermore, we compared the change in the interstitial adenosine level caused by ADA inhibition and nucleoside transport blockade, two well-established processes to stimulate the cell surface A adenosine receptors, in both thyroid states. A classical isolated organ technique was applied supplemented with the receptorial responsiveness method (RRM), a concentration estimating procedure. Via measuring the contractile force, the direct negative inotropic capacity of N-cyclopentyladenosine, a selective A receptor agonist, and methacholine, a muscarinic receptor agonist, was determined on the left atria isolated from 8-day solvent- and thyroxine-treated guinea pigs in the presence and absence of 2′-deoxycoformycin, a selective ADA inhibitor, and NBTI, a selective nucleoside transporter inhibitor. We found that ADA inhibition (but not nucleoside transport blockade) increased the signal amplification of the A adenosinergic (but not M muscarinergic) system. This action of ADA inhibition developed in both thyroid states, but it was greater in hyperthyroidism. Nevertheless, ADA inhibition produced a smaller rise in the interstitial adenosine concentration than nucleoside transport blockade did in both thyroid states. Our results indicate that ADA inhibition, besides increasing the interstitial adenosine level, intensifies the atrial A adenosinergic function in another (thyroid hormone-sensitive) way, suggesting a new mechanism of action of ADA inhibition. [ABSTRACT FROM AUTHOR]

We explored whether the M2 muscarinic receptor in the guinea pig ileum elicits a highly potent, direct-contractile response, like that from the M3 muscarinic receptor knockout mouse. First, we characterized the irreversible receptor-blocking activity of 4-DAMP mustard in ileum from muscarinic receptor knockout mice to verify its M3 selectivity. Then, we used 4-DAMP mustard to inactivate M3 responses in the guinea pig ileum to attempt to reveal direct, M2 receptor-mediated contractions. The muscarinic agonist, oxotremorine-M, elicited potent contractions in ileum from wild-type, M2 receptor knockout, and M3 receptor knockout mice characterized by negative log EC50 ( pEC 50) values ± SEM of 6.75 ± 0.03, 6.26 ± 0.05, and 6.99 ± 0.08, respectively. The corresponding Emax values in wild-type and M2 receptor knockout mice were approximately the same, but that in the M3 receptor knockout mouse was only 36% of wild type. Following 4-DAMP mustard treatment, the concentration–response curve of oxotremorine-M in wild-type ileum resembled that of the M3 knockout mouse in terms of its pEC 50, Emax, and inhibition by selective muscarinic antagonists. Thus, 4-DAMP mustard treatment appears to inactivate M3 responses selectively and renders the muscarinic contractile behavior of the wild-type ileum similar to that of the M3 knockout mouse. Following 4-DAMP mustard treatment, the contractile response of the guinea pig ileum to oxotremorine-M exhibited low potency and a competitive-antagonism profile consistent with an M3 response. The guinea pig ileum, therefore, lacks a direct, highly potent, M2-contractile component but may have a direct, lower potency M2 component. [ABSTRACT FROM AUTHOR]

Using the whole-cell voltage-clamp method, we examined an acute effect of dronedarone, a noniodinated benzofuran analogue of amiodarone, on Na+/Ca2+ exchange current ( I NCX) in guinea pig cardiac ventricular cells. The I NCX was recorded by ramp pulses with a holding potential of −60 mV using a pipette solution containing 226 nM free Ca2+ (20 mM 1,2-bis(2-aminophenoxy)ethane- N, N, N′, N′-tetraacetic acid and 10 mM Ca2+) and 20 mM Na+. The external solution contained 140 mM Na+, 1 mM Ca2+, and blockers of other currents and pumps such as Cs+, nifedipine, ryanodine, and ouabain. A selective potent NCX inhibitor, KB-R7943 (100 μM), was used to completely inhibit I NCX. Dronedarone inhibited I NCX in a concentration-dependent manner. The IC50 values for the outward and inward I NCX inhibition were about 33 and 28 μM, respectively, with the Hill coefficient of 1 for both. The inhibitory effect of dronedarone at 50 μM on I NCX did not change in the presence of trypsin in the pipette solution. Therefore, dronedarone is classified as a trypsin-insensitive NCX inhibitor and distinct from amiodarone which is a trypsin sensitive. We conclude that dronedarone inhibits I NCX but the potency is tenfold less than that of amiodarone. Dronedarone may modestly inhibit I NCX in a therapeutic concentration range. [ABSTRACT FROM AUTHOR]

The terminal 5-HT1B autoreceptors have attracted great pharmacological interest since they are potential targets for compounds modifying serotonergic neurotransmission. In the present work the in vivo biochemical properties of AR-A000002 ((R)-N-[5-methyl-8-(4-methylpiperazin-1-yl)-1,2,3,4-tetrahydro-2-naphthyl]-4-morpholinobenzamide), a novel selective 5-HT1B receptor antagonist, are reported. The effects of AR-A000002 on: 5-HT metabolism was measured as the ratio between 5-HIAA and 5-HT concentrations in different brain regions; 5-HT synthesis was measured as the accumulation of 5-HTP after inhibition of the aromatic amino acid decarboxylase activity with m-hydroxybenzylhydrazine (NSD1015); 5-HT release was measured using the microdialysis technique. 5-HT, 5-HIAA and 5-HTP concentrations were analyzed using high power liquid chromatography (HPLC) with electrochemical detection. AR-A000002 significantly enhanced 5-HT metabolism (5-HIAA/5-HT ratio) and 5-HT synthesis in guinea pig brain in the dose range 0.9–18 mg/kg s.c. (ED50=1 mg/kg s.c. in the four brain regions examined) with maximal effect seen after 2–4 h. AR-A000002 (9 mg/kg s.c.) significantly increased the extracellular concentrations of 5-HT and 5-HIAA by 20% in the guinea pig frontal cortex, measured with the in vivo microdialysis technique in freely moving guinea pigs. AR-A000002 (9 mg/kg s.c.) in combination with the 5-HT uptake inhibitor citalopram (5 mg/kg s.c.) increased the extracellular 5-HT concentration in guinea pig frontal cortex from 250 to 400% of the basal level. Citalopram alone decreased the extracellular 5-HIAA levels to 70% of the basal value. AR-A000002 counteracted the citalopram-induced decrease in 5-HIAA. Since the basal level of extracellular 5-HIAA was 160 times higher than that of 5-HT the 20% increase in 5-HIAA concentrations indicates that only a few percent of the exocytotically released 5-HT from the nerve terminals reached the extracellular space when the re-uptake mechanism was intact. The results also show that the terminal 5-HT1B autoreceptors are tonically activated under drug-free as well as citalopram conditions. The increase in plasma level of cortisol after AR-A000002 administration may indicate stimulation of post-synaptic 5-HT receptors. AR-A000002 also blocked 5-HT1B agonist-induced (CP-135,807) decrease in 5-HT metabolism and hypothermia (ED50=1 mg/kg s.c.), thus indicating competition between these two drugs. It is concluded that AR-A000002 is a 5-HT1B receptor antagonist that enhances the serotonergic neurotransmission in guinea pig brain. [ABSTRACT FROM AUTHOR]

Purpose. The chromanol HMR 1556 is a potent blocker of KvLQT1/minK potassium channels expressed in Xenopus oocytes. The compound is therefore a new class III antiarrhythmic drug with a distinct mechanism of action. However, the effect of HMR 1556 on atrial ion channels and the selectivity of block in the human heart has not been investigated. We tested the effects of HMR 1556 on repolarizing potassium currents in human and guinea pig atrial myocytes. Methods and results. Single atrial myocytes were isolated by enzymatic dissociation. Atrial potassium currents (IKs, IKr, in guinea pig, Ito, IKur, IK1 in humans) were recorded at 36°C in the whole cell mode of the patch clamp technique. HMR 1556 produced a concentration-dependent and reversible block of IKs with a half maximal concentration (EC50) of 6.8 nmol/l. 10 μmol/l HMR 1556 almost completely inhibited IKs (97.2±3.2%, n=6). Steady-state activation as well as kinetic properties of the current were not altered by HMR 1556. IKr currents were not affected up to concentrations of 10 μmol/l. HMR 1556 did not inhibit other potassium currents in human atrium: Ito, IKur and the classical inward rectifier potassium current IK1 were not significantly affected up to concentrations that completely blocked IKs (10 μmol/l). Conclusions. HMR 1556 is a highly-potent blocker of IKs channels without exerting effects on other potassium currents involved in atrial repolarization. Given the potential advantages of IKs vs. IKr blockade, the drug's new mechanism of action warrants further investigation to clarify its role as an antiarrhythmic agent. [ABSTRACT FROM AUTHOR]

We examined the in vivo actions of LY293111 sodium (2-[2-propyl-3-[3-[2-ethyl-4-(4-fluorophenyl)-5-hydroxyphenoxy]propoxy]phenoxy] benzoic acid sodium salt). Guinea pigs were used to evaluate the effect of this agent on (1) acute airway obstruction produced by intravenous leukotriene B4, (2) pulmonary granulocyte infiltration and delayed onset airway obstruction resulting from a 4-h leukotriene B4 inhalation and (3) lung inflammation after aerosol challenge with the divalent cationic ionophore A23187 (6S-[6α(2S*,3S*),8β(R*),9β,11α]-5-(methylamino)-2-[[3,9,11-trimethyl-8-[1-methyl-2-oxo-2-(1H-pyrrol-2-yl)ethyl]-1,7-dioxaspiro[5.5]undec-2-yl]methyl]-4-benzoxazolecarboxylic acid). Airway obstruction was quantitated using pulmonary gas trapping measurements and lung inflammation was evaluated by bronchoalveolar lavage (BAL) and histology. LY293111 sodium produced a dose-related inhibition of acute leukotriene B4-induced airway obstruction when administered i.v. (ED50=14 µg/kg) or p.o. (ED50=0.4 mg/kg). In contrast, LY293111 sodium did not inhibit the pulmonary gas trapping caused by aerosols of histamine, leukotriene D4, or the thromboxane mimetic U46619 (15 [(S)-hydroxy-11a,9a-(epoxymethano)prosta-5Z,13E-dienoic acid]). Oral LY293111 sodium inhibited leukotriene B4-induced bronchoalveolar lavage granulocyte infiltration and delayed onset airway obstruction at doses as low as 0.3 mg/kg. In A23187-challenged animals, pulmonary inflammation was markedly inhibited at 1 h, but not 2 h and 4 h post-exposure. We conclude that LY293111 sodium is a selective leukotriene B4 receptor antagonist with potent pulmonary anti-inflammatory activity. [ABSTRACT FROM AUTHOR]

Caffeine has been found to inhibit the formation and action of Ca2+-mobilizing inositol 1,4,5-trisphosphate (IP3) in pancreatic acinar cells. The aim of the present study was to investigate the effects of caffeine on cytoplasmic Ca2+ concentrations ([Ca2+]i) and amylase release in response to different agonists. [Ca2+]i was determined by cytofluorometry using fura-2 as indicator and amylase release with a substrate reagent. Stimulation with low concentrations of carbachol or cholecystokinin octapeptide (CCK-8) induces [Ca2+]i oscillations whereas higher concentrations cause sustained elevation of [Ca2+]i. The less efficacious agonists pilocarpine and CCK-JMV-180 evoke oscillations only. Caffeine inhibited carbachol-induced elevation of [Ca2+]i and amylase responses in a competitive manner, abolishing the responses to low and incompletely inhibiting the responses to high concentrations of the agonist. Also, the [Ca2+]i elevations by pilocarpine were abolished by caffeine. The effects on CCK-8-induced elevation of [Ca2+]i and amylase secretion were paradoxical, the caffeine inhibition being more pronounced at high than at low concentrations of CCK-8. This enigma was further emphasized by moderate effects of caffeine on the responses to CCK-JMV-180. The results indicate that carbachol, pilocarpine and high concentrations of CCK-8 elicit IP3-mediated responses and that CCK-JMV-180 and low concentrations of CCK-8 elevate [Ca2+]i and stimulate amylase release by another signal transduction mechanism. [ABSTRACT FROM AUTHOR]

The effect of the phosphodiesterase inhibitor 4-(3-cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone (ZK 62711) on gastric secretion and the cyclic AMP system of the gastric mucosa was studied in rats and guinea pigs. In rats, 0.03-0.3 μmoles/kg ZK 62711 i.v. stimulated acid and pepsin secretion in a dose-dependent manner and 0.03 μmoles/kg i.v. enhanced the effect of histamine. In guinea pigs no reproducible stimulation was found after intravenous injections of ZK 62711. The stimulation of gastric secretion in the rat by 0.3 μmoles/kg ZK 62711 i.v. was not affected by vagotomy but was totally inhibited by 10 μmoles/kg cimetidine i.v. The structurally related phosphodiesterase inhibitor, 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidine (Ro 20-1724), at the dose of 3.3 μmoles/kg i.v. stimulated gastric secretion in anaesthetised rats to a similar extent as 0.3 μmoles/kg ZK 62711 i.v. The content of cyclic AMP in the rat gastric mucosa in vivo was slightly increased by 10 μmoles/kg ZK 62711 i.v, whereas lower doses did not change it. Accumulation of cyclic AMP in the rat gastric mucosa by 50 μmoles/kg histamine i.v. was enhanced by simultaneous injections of 3.3 μmoles/kg ZK 62711 i.v. In rat gastric tissue slices in vitro 1-50 μM ZK 62711 increased the level of cyclic AMP but 100 μM histamine had no effect in the absence or presence of ZK 62711. In gastric mucosal slices of the guinea pig 10 and 50 μM ZK 62711 increased the cyclic AMP content which effect was inhibited by 100 μM cimetidine and enhanced the stimulatory effect of 100 μM histamine on cyclic AMP. The activity of soluble cyclic AMP phosphodiesterase was inhibited by ZK 62711 in the rat (IC=18 μM) and guinea pig gastric mucosa (IC=1.5 μM). Adenylate cyclase was not affected in the homogenate of the guinea pig gastric mucosa. The results indicate that the phosphodiesterase inhibitor ZK 62711 which increases cyclic AMP levels in the gastric mucosa in vivo and in vitro, is a potent stimulator of gastric acid secretion. [ABSTRACT FROM AUTHOR]

The presence and distribution of myocardial α-adrenoceptors in different parts of the heart of various mammalian species was investigated. For this reason experiments were performed in isolated cardiac preparations of rats, guinea pigs and cats. In order to obtain more information about the nature of the cardiac α-adrenoceptors additional experiments were undertaken at different temperatures. These studies were aimed to show whether or not a conversion of β-to α-adrenoceptors or vice versa takes place. Moreover, we analyzed the influence of hypothyroidism on the sensitivity of α- and β-adrenoceptors of preparations from rats fed with propylthiouracil. Finally, we tried to find out whether stimulation of these α-adrenoceptors leads to the formation of the cyclic nucleotides cAMP and cGMP. The following results were obtained: [ABSTRACT FROM AUTHOR]

In isolated epithelia of guinea pig jejunum the transcellular permeation of 10 M (carboxyl-C)-3,4,5-trimethoxybenzoic acid (TMBA) in the direction blood-lumen was more than 10 times greater the transcellular permeation of 10 M (carboxyl-was reduced to less than 2 by anaerobiosis or by increasing TMBA concentrations of up to 10 M. Under aerobic conditions the cellular uptake of TMBA (10 M) from the blood side was twice as high as that from the lumen side. In anaerobiosis the percentage of TMBA taken up into the epithelium was enhanced, when TMBA was administered on the lumen side, while the percentage was unchanged after administration on the blood side; thereby the difference in cellular TMBA concentrations was abolished. Similar results were obtained under aerobic conditions, if the TMBA concentration was increased up to 10 M. The results are consistent with a three-compartment model with an intermediate compartment distinguished by a high pH as compared to that of the outer compartments and by a luminal boundary highly permeable for the ionized form of the substrate in contrast to the contraluminal boundary. [ABSTRACT FROM AUTHOR]

Cumulative dose-response curves for histamine were determined on acid secretion from the isolated guinea pig gastric mucosa. Two H-receptor antagonists-metiamide and cimetidine-behaved like competitive antagonists to histamine on gastric acid secretion in vitro. The isolated guinea pig gastric mucosa seems to be a suitable in vitro model for analysing the action of compounds on receptors involved in acid secretion. [ABSTRACT FROM AUTHOR]

After administration of the anorectic drug fenfluramine (40 mg/kg per day) to rats (s. c. or i. p.) and guinea pigs (p.o.) over periods of 2 to 11 weeks, electron microscopic examination was performed on lungs, liver, lymphatic tissues and peripheral blood of both species, and on adrenal glands and ovaries of rats. In rats, fenfluramine caused the following alterations (listed according to the duration of treatment): Formation of abnormal lamellated inclusions in lymphocytes and plasma cells of lymphatic tissues, appearance of 'foam cells' in lung alveoli; formation of abnormal lamellated inclusions in corpus luteum, lymphocytes of peripheral blood, and in adrenal cortex. In guinea pigs, the same alterations were found in lymphocytes of spleen and peripheral blood, in hepatocytes and in lung. The present observations support the concept of a generalized phospholipidosis induced by amphiphilic compounds. The potency of fenfluramine to induce a lipidosis is, however, considerably less pronounced than that previously demonstrated for the anorectic drug chlorphentermine. This difference is suggested to be due mainly to the lower degree of amphiphilia of fenfluramine. [ABSTRACT FROM AUTHOR]

Experiments were performed on isolated electrically driven and spontaneously beating atria from guinea pigs and rabbits in order to characterize the cardiostimulatory effect of phenylephrine as either β- or α-sympathomimetic. [ABSTRACT FROM AUTHOR]

Sigma receptors are found in motor and limbic areas in the brains of humans, non-human primates, and rodents. The most extensive pharmacological studies of ligand binding to sigma receptors have utilized brain tissue from guinea pigs, where two subtypes of sigma receptor, designated sigma1 and sigma2, have been identified. Few functional roles for sigma receptors have been described. Their location in guinea pig striatum, a terminal field of dopaminergic projections arising from the substantia nigra, suggested that this tissue would be a logical choice in which to examine physiological properties of sigma receptor activation. We found that sigma1 receptor agonists inhibited N-methyl- d-aspartate-stimulated [3H]dopamine release from guinea pig striatal slices in a concentration-dependent manner. The inhibition by sigma1 receptor agonists was reversed by a selective sigma1 receptor antagonist, as well as by a non-subtype-selective sigma receptor antagonist. The ability of agonists working through sigma1 receptors, but not through sigma2 receptors, to inhibit the stimulated release of catecholamines appears to be a unique characteristic of guinea pig striatum. We have previously reported that in rat striatum and hippocampus, as well as in guinea pig nucleus accumbens, prefrontal cortex, and hippocampus, activation of either sigma receptor subtype inhibits such release. Stimulated release of [3H]dopamine from guinea pig striatum was also inhibited by the phencyclidine receptor agonist dizocilpine, but this inhibition was not reversed by the sigma receptor antagonists. Therefore, the inhibition produced by sigma receptor agonists was not mediated via the phencyclidine binding site within the N-methyl- d-aspartate-operated cation channel. Our findings support the hypothesis that sigma receptor activation provides a mechanism of modulating dopamine release from striatum, and that striatal tissue from guinea pigs appears to be an appropriate model for characterizing sigma1 receptor-mediated effects. [ABSTRACT FROM AUTHOR]

Terodiline, an anticholinergic drug with a Ca2+ blocking action, is thought to be associated with torsade de pointes, a serious ventricular tachycardia. NS-21 is a newly developed drug for the treatment of urinary frequency and urinary incontinence and it has pharmacological properties similar to those of terodiline. It remains unknown, however, whether NS-21 and its active metabolite, RCC-36, have any proarrhythmic activity. The electrophysiological properties of NS-21 and RCC-36 were examined in guinea pig ventricular myocytes and were compared with those of terodiline using the whole-cell patch-clamp technique. NS-21, RCC-36 and terodiline inhibited L-type Ca2+ currents in a concentration-dependent manner with IC50 values of 27.0, 27.0 and 33.5 μM, respectively. At a concentration of 10 μM, terodiline inhibited both the time-dependent current and the tail current of the delayed rectifier K+ current, with the latter being significantly inhibited at voltages more positive than +10 mV. In contrast, NS-21 and RCC-36 had almost no effect on either of these currents. Terodiline also inhibited the inward rectifier K+ current significantly at voltages more negative than -100 mV, whereas NS-21 and RCC-36 had little effect. If the proarrhythmic activity of terodiline resulted primarily from the combined inhibition of K+ and Ca2+ currents, one might expect that NS-21 and RCC-36, which inhibit L-type Ca2+ currents without affecting either the delayed rectifier K+ current or the inward rectifier K+ current, would not share the proarrhythmic activities of terodiline. [ABSTRACT FROM AUTHOR]

We determined and compared the molecular properties of histamine H1 receptor binding sites in bovine thoracic aorta smooth muscle and guinea pig myocardial membranes from ventricles with saturation and inhibition binding assay, using 3H-mepyramine to label the receptor and specific and selective H1 receptor agonists of the 2-phenylhistamine group as displacers of specific 3H-mepyramine binding. 3H-mepyramine binds in a saturable manner to a homogenous population of binding sites with a K D of 5.6 nM and a B max of 57 fmol/mg of protein in bovine aorta vascular smooth muscle membranes, whereas in the guinea pig myocardium high and low affinity 3H-mepyramine binding sites exist having the following molecular characteristics: a K D of 1.6 nM and a B max of 99 fmol/mg of protein (high affinity site) and a K D 15.0 nM and a B max of 466 fmol/mg of protein (low affinity site). Halogenated 2-phenylhistamines: 2-(3-fluoro-) (2-FPH), 2-(3-trifluoromethyl-) (2-triFMPH), 2-(3-chloro-) (2-CPH), 2-(3-bromo-) (2-BPH) and 2-(3-iodophenyl)histamine (2-IPH), which showed high selectivity and potency for H1 receptors in the functional pharmacological studies, were potent inhibitors of specific radioligand binding in comparison with histamine and parent nonhalogenated 2-phenylhistamine (2-PH). Their rank order of potencies and affinities differ significantly for the vascular and cardiac H1 receptor binding sites: Specific 3H-mepyramine binding to H1 receptors in bovine vascular smooth muscle membranes was displaced in a biphasic manner by 2-(3-fluoro-), 2-(3-trifluoromethyl-), 2-(3-chloro-), 2-(3-bromo-), 2-(3-iodophenyl)histamine and histamine. In guinea pig ventricular myocardium the rank order was 2-(3-iodo-), 2-(3-bromo-), histamine, 2-(3-chloro-), and 2-(3-fluorophenyl)histamine showing better correlation with the lipophilicity of the derivatives than in vascular tissue (order of lipophilicity: 2-triFMPH >2-IPH >2-BPH >2-CPH >2-FPH >>2-PH). Displacement of the radioligand binding to myocardial H1 receptor by the above drugs is (except for 2-(3-fluorophenyl)histamine), better fitted to a two-site model. 2-phenylhistamine, which acted as a moderate agonist in functional studies, displaced the radioligand in a monophasic manner and was the weakest displacer of specific radioligand binding in both model systems ( pK i = 5.76 – vascular and 5.57 – cardiac tissue). The agonistic nature of the halogenated 2-phenylhistamine derivatives was confirmed on the molecular level, since their interaction with the H1 receptor is regulated by guanine nucleotides. GTP (0.1 mM) significantly lowered the affinities of all tested halogenated 2-phenylhistamines and histamine for H1 receptor binding site converting biphasic displacement curves, to monophasic ones, whereas GTP had no effect on the affinity of 2-PH. The results of this study support the conclusions that bovine vascular and guinea pig myocardial histamine H1 receptors differ in their molecular properties. Selective and potent H1 receptor agonists of 2-phenylhistamine class can discriminate between vascular and cardiac receptor. [ABSTRACT FROM AUTHOR]

Guinea pig inferior vena cava contracted in response to leukotriene (LT)C, LTD, LTE U46619, phenylephrine, histamine, and KCl. Although LTC, LTD, and U46619 were the most potent agonists, active tension generated by these eicosanoids was only about half that of histamine or KCl. LTE and phenylephrine were marginally active. Biochemical analysis showed vena cava able to convert about 23% LTC to LTD and LTE in 45 min. Pretreatment with acivicin prevented this by abrogating conversion of LTC to LTD. A subthreshold concentration of LTE reduced responses to LTC and LTD. LY171883 and WY-48252 competitively antagonized LTD-induced contractions of vena cava. In contrast, these antagonists blocked contractions to LTC in a biphasic manner. Lower segments of the LTC concentration-response curves were less affected than the upper portion suggesting the possibility of 2 LTC receptor subtypes. Our results indicate that LTE is a weak or partial agonist in this tissue and furthermore they suggest a lack of high affinity receptors for LTE favoring LTC and LTD. Indomethacin did not influence contractions to the leukotrienes or histamine. However, the response to U46619 was greatly enhanced suggesting release of a vasodilator prostaglandin as part of the overall response of the vena cava to the thromboxane A2 mimetic. [ABSTRACT FROM AUTHOR]

Inhibition of [H]-5-hydroxytryptamine ([H]-5-HT) release from guinea pig brain slices via activation of the terminal 5-HT autoreceptor has previously been characterised as a model of 5-HT receptor activation, based on the rank potencies of a range of agonists, and the potent antagonism of the inhibitory effects of 5-HT by metitepine. The present study uses this model, in slices of the guinea pig hippocampus, to examine the antagonist potency of metitepine against the 5-HT receptor agonists sumatriptan, 5-carboxamidotryptamine (5-CT) and 5-HT Addition of metitepine to the perfusion buffer (30, 300 and 1000 nmol/l) significantly shifted the concentration-response curve to 5-HT, producing a Schild slope of 1.1, and a pA value of 7.6. However, the ability of metitepine to antagonise the effects of sumatriptan or 5-CT in this model was less marked. A clear-cut shift in the concentration-response curve to sumatriptan was only achieved at1000 nmol/l metitepine (apparent pA = 6.7),and this was similar to the ability of metitepine to attenuate the effects of 5-CT (apparent pA 7.0 at 300 nmol/l and 6.7 at 1000 nmol/l). These findings suggest heterogeneity in the receptor mediating inhibition of [H]-5-HT release in guinea pig hippocampus. [ABSTRACT FROM AUTHOR]

The effects of histamine on delayed K current (I) were investigated in patch-clamped single guinea pig ventricular myocytes. Histamine increased I with a maximal fractional response of 2.7 and a k of 9.4 × 10 mol/l. At a concentration of 10 mol/l, histamine did not increase I significantly, but increased I by 52% ± 12%. The voltage-dependence of I activation, the reversal potential and the time course of the I tail decay were not changed by histamine. Under pretreatment with 10 mol/l of ranitidine, neither histamine (10 mol/l) nor 2-pyridylethylamine (10 mol/l) caused any sizable increase in I. When the cell was pretreated with a saturating dose of isoproterenol (10 mol/l), histamine did not additively enhance I. The I enhancement by 3 × 10 mol/l histamine was partially antagonized by concurrent exposure to 5 × 10 mol/l carbachol. Whereas, use of a higher concentration of histamine (10 mol/l) obscured the inhibitory effect of carbachol. It is concluded that histaminergic action of I is attributed exclusively to H receptor-mediated reactions involving G protein and adenylate cyclase. [ABSTRACT FROM AUTHOR]

Field stimulation (0.125-10 Hz) of longitudinal smooth muscle strips taken from the fundus of guinea pig stomach caused frequency related contraction responses (atropine-sensitive) associated with relaxation at the higher frequencies (both responses tetrodotoxin-sensitive). Dopamine, noradrenaline, adrenaline and apomorphine antagonised the contraction responses at all frequencies; the concentration-response curves were steep and the use of higher concentrations was precluded by changes in base line tension per se. That noradrenaline was approximately 10 fold more potent than dopamine, that the dopamine response was not antagonised by the dopamine antagonists haloperidol, domperidone or (-)sulpiride, but was mimicked by the α-agonist guanfacine and partially antagonised by the α-adrenoceptor antagonist yohimbine (which could also antagonise the inhibitory actions of guanfacine) would indicate that a component of dopamine's action to reduce cholinergic activity is effected via α-adrenoceptor mechanisms. A failure of phenylephrine to mimic, or prazosin to attenuate the agonist inhibitory responses would not indicate an additional α-adrenoceptor involvement. The persistence of the dopamine response following disruption of noradrenergic function by reserpine or inhibition of catecholamine reuptake processes (desmethylimipramine plus corticosterone, GBR 13098) would indicate a direct action of dopamine. It is concluded that the ability of dopamine to reduce cholinergic induced contractions in longitudinal smooth muscle of the stomach does not reflect a dopamine receptor or α-adrenoceptor stimulation but may involve an action on α-adrenoceptors. [ABSTRACT FROM AUTHOR]

The term 'allergen tachyphylaxis' (AT) describes a progressively decreased bronchial reactivity to allergen exposition after repeated allergen challenge [in our test system measured by sequential inhalative antigen challenge of sensitized guinea pigs (GP)]. The hypothesis that AT is mediated by endogenous prostaglandin E (PGE) biosynthesis was tested in vivo on GP sensitized to ovalbumin (OA). Different groups of animals were challenged with simultaneous inhalation of OA (repeatedly at time 0, 10, 20, 60 and 70 min) together with inhibitors of PGE-biosynthesis (parachloromercurobenzoic acid = PCMB, copper sulfate = Cu, copper dithiotreitol complex = CuDTT, dithiotreitol = DTT and dimercaptopropanol = DMP) or agents increasing PGE production (aurothioglucose = Au, zinc dithiotreitol complex = ZnDTT and reduced glutathion = GSH). Bronchial obstruction was measured by whole body plethysmography. PCMB, Cu, CuDTT, DTT and DMP inhibited AT, whereas Au and ZnDTT enhanced AT. Acetylsalicylic acid (ASA) treatment prevented AT. Aerosols of PGE, but not of prostacyclin or prostaglandin D restored AT in ASA treated animals. In addition to these in vivo experiments in vitro investigations showed that PCMB, Cu and DTT decreased while ZnDTT increased PGE biosynthesis of allergen challenged GP lungs. It is concluded that AT, an important self-protecting mechanism of GP bronchial asthma, is mediated at least in part via endogenous PGE. [ABSTRACT FROM AUTHOR]

Ovalbumin (OA) sensitized guinea pigs were repeatedly challenged with 1% OA in saline nebulized ultrasonically at the 0, 10, 20, 60 and 70th min. The intensity of bronchial obstruction was measured by body plethysmography. The first three challenges (0, 10, 20 min) caused strong asthmatic reactions in all animals, the last two (60, 70 min) only mild ones in 10 out of 15 animals. The development of this 'tachyphylaxis' was markedly reduced by pretreatment of the animals with cyclooxygenase inhibitors (indomethacin 10 mg/kg intraperitoneally resp. acetylsalicylic acid 10 mg/kg orally 2h before test). The effect of both inhibitors (i.e. inhibition of tachyphylaxis) was abolished by supplementing prostaglandin E as aerosol simultaneously to the allergen (100-200 ng per inhalation). The results suggest that allergen tachyphylaxis we have observed in vivo might be due to synthesis of cyclooxygenase products, e.g. prostaglandin E. [ABSTRACT FROM AUTHOR]

N-Demethylation of C-aminopyrine (C-AP), labelled at the methyl groups of the tertiary amino group, yields HCHO and C-monomethylaminoantipyrine (C-MMAAP) which also undergoes N-demethylation, however, at a slower rate as measured in hepatic microsomes. As after intraperitoneal application to male guinea pigs of C-AP (75 mg/kg; 50 μCi/kg), exhalation rate of CO declines in a biphasic manner, the hypothesis was tested whether the terminal part might reflect N-demethylation of MMAAP. The application of C-MMAAP (70 mg/kg; 10 μCi/kg), resulted in monophasic curves of CO exhalation rate. Their half lives were, however, longer than terminal half lives obtained after C-AP. Obviously, this terminal phase does not represent CO formation from the metabolite MMAAP only, but C-AP might still contribute to CO production. Confirmation was obtained by HPLC determination of AP and MMAAP in serum after injection of AP. Shortly after injection, high concentrations of AP and low ones of MMAAP were found in blood from the portal vein and systemic circulation. Thus, initial parts of CO-exhalation rate curves reflect predominantly AP metabolism whereas later phases provide hybrid information. [ABSTRACT FROM AUTHOR]

Renal and extrarenal effects of aldosterone antagonists on potassium balance were investigated in guinea pigs. It is concluded that in guinea pigs aldosterone antagonists favour a shift of potassium from the extra- to the intracellular space by a direct effect on cardiac and skeletal muscle. [ABSTRACT FROM AUTHOR]