Calcium Dependence of Contractile Activation of Isolated Sheep Urethra II: Responses to Exogenous Noradrenaline

Isolated smooth muscle of sheep urethra responded to exogenous noradrenaline (NA) with a concentrationdependent contraction. After exposing the preparations for 30 min. to calcium-free medium, NA in a submaximal concentration was still able to produce a contractile response amounting to 36% of control value in calcium-containing solution. Readmission of calcium (administered cumulatively) restored the response in a concentration-related fashion to 85% (at 5 mM calcium) of control level. Nifedipine and verapamil failed to inhibit these graded calcium contractions in the presence of NA. Verapamil and diltiazem also failed to significantly prevent contraction induced by NA in a submaximum concentration, but nifedipine showed a concentration-dependent inhibitory effect, with a potency 1000 times lower than that observed on K⁺ (124 mM) induced contractions. In calcium-free medium, repeated applications of NA at 30 min. intervals, induced a progressive reduction in contractile amplitude. Nifedipine or verapamil did not affect the time course for recovery of NA-induced contraction when the preparations were returned to calcium containing medium, but the recovery was blocked almost completely by lanthanum. Furthermore, lanthanum abolished the remaining NA contraction in calcium-free medium. The results suggest that NA-induced contraction in isolated sheep urethra is dependent on both influx of extracellular calcium and on release of intracellular calcium. Calcium influx for contractile activation and refilling of intracellular stores seem to occur through membrane channels that can only partly be blocked with calcium antagonists.     

Calcium dependence of contractile activation of isolated sheep urethra. I: Responses to electrical stimulation

In isolated sheep urethral smooth muscle at resting tension, responses elicited by electrical stimulation of nerves were either contraction (40%), or contraction preceded by relaxation (60%). All responses were suppressed by tetrodotoxin (TTX), but at stimulation frequencies exceeding 16 Hz, there was a small TTX resistant contraction, which at the highest frequency used (50 Hz) amounted to about 15% of the total response. alpha-Adrenoceptor blockade suppressed the contractions, as did chemical sympathectomy with 6-hydroxydopamine. In calcium-free medium all electrically induced contractions were abolished. Nifedipine, verapamil, and lanthanum had concentration-dependent depressant actions, whereas Bay K 8644 (10(-6) M) significantly increased the responses. In preparations prelabelled with 3H-noradrenaline, electrical stimulation caused a release of 3H that could be blocked by TTX, and effectively reduced by calcium-free medium and lanthanum. However, the release was unaffected by nifedipine, and moderately reduced by high concentrations of verapamil (10(-5) M). It is suggested that contractile responses to electrical stimulation in isolated sheep urethral smooth muscle are mediated by the sympathetic nervous system, mainly through release of noradrenaline stimulating postjunctional alpha 1-adrenoceptors. The release of noradrenaline, as well as the action of released noradrenaline on postjunctional membranes are calcium-dependent. However, the calcium entry pathways used are partly different from those inhibited by organic calcium antagonists.     

Calcium Dependence of Contractile Activation of Isolated Sheep Urethra I: Responses to Electrical Stimulation

In isolated sheep urethral smooth muscle at resting tension, responses elicited by electrical stimulation of nerves were either contraction (40%), or contraction preceded by relaxation (60%). All responses were suppressed by tetrodotoxin (TTX), but at stimulation frequencies exceeding 16 Hz, there was a small TTX resistant contraction, which at the highest frequency used (50 Hz) amounted to about 15% of the total response. α-Adrenoceptor blockade suppressed the contractions, as did chemical sympathectomy with 6-hydroxydopamine. In calcium-free medium all electrically induced contractions were abolished. Nifedipine, verapamil, and lanthanum had concentration-dependent depressant actions, whereas Bay K 8644 (10^?6 M) significantly increased the responses. In preparations prelabelled with³H-noradrenaline, electrical stimulation caused a release of ³H that could be blocked by TTX, and effectively reduced by calcium-free medium and lanthanum. However, the release was unaffected by nifedipine, and moderately reduced by high concentrations of verapamil (10^?5 M). It is suggested that contractile responses to electrical stimulation in isolated sheep urethral smooth muscle are mediated by the sympathetic nervous system, mainly through release of noradrenaline stimulating postjunctional α₁-adrenoceptors. The release of noradrenaline, as well as the action of released noradrenaline on postjunctional membranes are calcium-dependent. However, the calcium entry pathways used are partly different from those inhibited by organic calcium antagonists.     

Comparison of the effects of hydralazine and nifedipine on contractions and 45Ca influx of rat aorta.

The effect of the vasodilator hydralazine has been compared with nifedipine on KCl-(K+) (60 mM) and noradrenaline-(NA) (10 microM) induced 45Ca uptake and contractile responses in rat aorta arterial strips without endothelium. Hydralazine (0.5-10 mM) was equally effective in relaxing K(+)- (IC50 = 2.2 +/- 0.17 mM) and NA- (IC50 = 3.06 +/- 0.25 mM) induced tension, the degree of relaxation depending on the dose. Nifedipine totally inhibited K(+)- (IC50 = 3.16 +/- 0.28 nM) induced contractions with lower doses than were necessary to relax (up to 54.0 +/- 4.1% with supramaximal concentrations) NA-induced contractions (IC50 = 1.48 +/- 0.12 microM). In the experiments in a calcium-free medium, nifedipine (1 microM) had no effect on the NA- (10 microM) induced contractions whereas hydralazine (1 mM) strongly inhibited them. Nifedipine did not affect the basal uptake of 45Ca but the induced uptakes were reduced to 66.3 +/- 3.2% (K+) and 65.5 +/- 4.1% (NA) of their basal values. Hydralazine did not affect the basal uptake of 45Ca nor that induced by the two vascoconstrictor agents. These results suggest that nifedipine acts on the cell membrane by blocking the movements of calcium through the voltage-dependent and receptor-operated calcium channels, whilst hydralazine has an intracellular effect.     

Ryanodine- and cyclopiazonic acid-sensitive components in human vas deferens contractions to noradrenaline

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Calcium influx and protein kinase C activation involved in uterine vasoconstriction in guinea pigs

The mechanical responses of circular segments of uterine arteries to different combinations of vasoactive agents and putative inhibitors of calcium fluxes were examined using a sensitive in vitro method. Exposure to high potassium (127 mM) or to noradrenaline (NA; 0.1 mM) resulted in a rapid, initial increase in tension of the vascular preparation to reach a sustained level of contraction (approximatively 12 mN) which lasted for at least 15 min. The protein kinase C activator, 4-beta-phorbol-12,13-esther-dibutyrate (10 microM), induced a slowly developing but sustained contractile response with a maximum (PDBumax) of only 4 mN. Addition of the calcium ionophore, A23187, to PDBu-contracted vessels did not increase tension, while the maximum tension evoked by A23187 per se (3 microM) was 5 mN. Ionomycin had only a small contractile effect on the uterine artery. The contraction evoked by depolarization (potassium) or alpha 1-adrenoceptor stimulation (NA) was decreased in nominally calcium-free medium containing EDTA (0-100 microM) or EGTA, while uterine arteries loaded intracellularly with the calcium chelator, quin-2, responded to the vasoconstrictors almost as well as the control preparations. Blockade of calcium influx with Cd2+ (greater than 0.01 mM), nifedipine (greater than 3 microM), verapamil (greater than 1 microM) and TMB-8 (greater than 10 microM) reduced the tension evoked by potassium somewhat more than it reduced the contractions induced by NA, while the opposite was seen in the presence of Ni2+ (greater than 0.1 mM). Inhibition of calmodulin-dependent enzymes by W7 (greater than 10 microM) reduced the maximum tension evoked by potassium and NA to a similar extent.(ABSTRACT TRUNCATED AT 250 WORDS)     

Extracellular and intracellular calcium sources mediating contractile responses of smooth muscle in bovine ovarian follicle and ovarian artery

The relative importance of extracellular and intracellular calcium sources mediating smooth muscle contraction in ovarian follicle and ovarian artery was assessed in experiments on the influence of nifedipine, D-600, amrinone, diethylstilbestrol (DES), lanthanum and/or calcium removal on contractions induced by K+ depolarization, by noradrenaline, histamine and acetylcholine. The K+-induced response was biphasic in the ovarian artery but not in the ovarian follicle. The K+-induced contraction in both preparations was greatly inhibited by nifedipine (1 microM), D-600 (10 microM) and lanthanum (2 mM). Although both phases of the responses in the ovarian artery appeared to be completely dependent on extracellular calcium, phase I was significantly more sensitive to nifedipine than phase II. Incubation in calcium-free medium for 15 min almost abolished the K+-induced contraction. Noradrenaline- and histamine-induced contractions of ovarian follicle were essentially unaffected by nifedipine (1 microM) and D-600 (10 microM) whereas the noradrenaline-induced contraction in ovarian artery was inhibited significantly by D-600 (1 and 10 microM) but not nifedipine (1 microM). In calcium-free medium containing EGTA (1 mM) the responses of ovarian follicle to noradrenaline and histamine were reduced by 26 and 22% respectively. When preparations were stimulated with noradrenaline more than one in calcium-free medium, the contraction decreased progressively compared to time-matched controls. The response was 34% of the control after 50 min in calcium-free medium containing EGTA. In the ovarian artery the response obtained (6% of control) was significantly smaller (P less than 0.05) than that in the follicle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of niflumic acid on noradrenaline-induced contractions of the rat aorta.

1. The effects of niflumic acid, an inhibitor of calcium-activated chloride channels, were compared with the actions of the calcium channel antagonist nifedipine on noradrenaline-evoked contractions in isolated preparations of the rat aorta. 2. The cumulative concentration-effect curve to noradrenaline (NA) was depressed by both nifedipine and niflumic acid in a reversible and concentration-dependent manner. The degree of inhibition of the maximal contractile response to NA (1 microM) produced by 10 microM niflumic acid (38%) was similar to the effect of 1 microM nifedipine (39%). 3. Contractions to brief applications (30 s) of 1 microM NA were inhibited by 55% and 62% respectively by 10 microM niflumic acid and 1 microM nifedipine. 4. In the presence of 0.1 microM nifedipine, niflumic acid (10 microM) produced no further inhibition of the NA-evoked contractions. Thus, the actions of niflumic acid and nifedipine were not additive. 5. In Ca-free conditions the transient contraction induced by 1 microM NA was not inhibited by niflumic acid (10 microM) and therefore this agent does not reduce the amount of calcium released from the intracellular store or reduce the sensitivity of the contractile apparatus to calcium. 6. Niflumic acid 10 microM did not inhibit the contractions produced by KCl (up to 120 mM) which were totally blocked by nifedipine. Contractions induced by 25 mM KCl were completely inhibited by 1 microM levcromakalim but were unaffected by niflumic acid. 7. It was concluded that niflumic acid produces selective inhibition of a component of NA-evoked contraction which is probably mediated by voltage-gated calcium channels. These data are consistent with a model in which NA stimulates a calcium-activated chloride conductance which leads to the opening of voltage-gated calcium channels to produce contraction.     

Cellular calcium and the contraction induced by prostaglandin F2 alpha in feline cerebral arteries

The influences of different calcium-entry blockers, sialidase and caffeine on the biphasic contraction induced by prostaglandin (PG) F2 alpha in the feline basilar artery (BA) were studied in calcium-free medium. After incubation in calcium-free solution, PGF2 alpha induced a contraction of the BA amounting to 87% of the contraction in calcium-containing solution. The response was biphasic in 41 out of 42 vessel segments. PGF2 alpha-induced contractions were markedly attenuated in TRIS-buffered solutions as compared to contractions in Krebs solution. PGF2 alpha failed to induce a biphasic contraction (8 out of 9 preparations) in calcium-free HEPES-buffered solution. Calcium entry blockade with 1 mM manganese or 10(-5) M diltiazem abolished the second and major phase of the PGF2 alpha-induced contraction in calcium-free Krebs solution. The second contraction phase was also eliminated in four out of five preparations pretreated with sialidase (1 unit/ml for 30 min.), but was unaffected by a brief exposure to 20 mM caffeine in calcium-free medium. The present findings strongly support previous suggestions that a major part of the PGF2 alpha-induced contraction in calcium-free medium is mediated via the release of calcium bound to the exterior aspect of the cell membrane.     

The Effect of Nifedipine on Isolated Human Peripheral Vessels

Abstract Isometric tension was recorded in ring preparations of human peripherial arteries and veins contracted by potassium (127 mM) and noradrenaline (1.8 × 10^-5 M). In the veins, nifedipine had a marked relaxing effect on contractions induced by both agents, and also reduced the contractile amplitude when added prior to stimulation. The inhibiting effect of nifedipine was more marked on the potassium than on the noradrenaline-evoked responses. This was in contrast to verapamil, which inhibited the noradrenaline-induced contractions significantly more, than those produced by potassium. After immersion of the vein preparations in calcium-free medium for 30 min., the potassium contracture decreased to 26±2.0% (mean ± S.E.M.) of the response in normal Krebs solution, and the noradrenaline-evoked response to 7.1±0.8%. The responses to both agents were completely restored when the calcium concentration was increased from 0 to 4 mM. Nifedipine (1.5 × 10^-7 M) depressed the potassium contracture in calcium-free solution to 7.3±1.6%, and the noradrenaline response to 5.5±1.6%; on addition of calcium, the response elicited by potassium increased to 16±1.7%, and that by noradrenaline to 56±8.6%. Compared with its actions on the veins, the effect of nifedipine on the arterial preparations was less pronounced. In the arteries, too, the inhibiting effect of nifedipine was significantly more pronounced on the potassium than on the noradrenaline-induced contraction. Immersion for 30 min. in calcium-free medium reduced the response to potassium to 61±6.0% and that to noradrenaline to 68±5.6% of the control in normal Krebs. Nifedipine (2.9 × 10^-7 M) further reduced the potassium contraction to 20±4.0%; the response to noradrenaline was unaffected, being 74±6.4% of the control.     

Contractile Effects of Some Polypeptides on the Isolated Urinary Bladder of Guinea-pig,Rabbit, and Rat

Abstract: In isolated detrusor from guinea-pigs and rats substance P (SP) induced concentration-dependent phasic contractions. Rabbit detrusor strips responded to SP with an initial phasic and an ensuing tonic contraction. The concentration-response curve to SP in this preparation had a biphasic appearance. Eledoisin (E) caused contractile responses similar to those of SP when tested on the guinea-pig detrusor. Somatostatin was tested on the rabbit urinary bladder; it caused a concentration-dependent rise in basal tone, but no phasic contraction. The responses to SP and E were not affected by tetrodotoxin or physostigmine. They were only partly inhibited by high concentrations of atropine and the anticholinergic drug PR 197. Noradrenaline and isoprenaline caused a concentration-dependent inhibition of the peptide induced responses in guinea-pig bladder; at high concentrations of the amines this inhibition was almost complete. Indomethacin did not affect the SP induced contractions in the guinea-pig bladder, reduced the responses in the rabbit detrusor, but increased them in the rat bladder. Contractions elicited by SP and E were rapidly diminished or abolished after 30 min. treatment in a calcium-free medium. The responses were also inhibited by the calcium antagonist nifedipine. In guinea-pig preparations depolarized by potassium (127 mM) both SP and E caused a contractile response approximately 20% of that obtained in normal Tyrode solution. It is concluded that SP and E cause contraction of detrusor by a direct effect on the smooth muscle cells, and that this response is dependent on the extracellular calcium concentration. Prostaglandins may be involved in the SP-induced response of the rabbit detrusor.     

Inhibitory effects of nitrendipine on myometrial and vascular smooth muscle in human pregnant uterus and placenta

Human myometrial visceral and vascular preparations and placental chorionic and stem villous vessels were dissected from myometrial and placental specimens obtained at term Caesarean section and after vaginal delivery. Vascular ring preparations and myometrial strips were mounted in organ bath and isometric tension recorded. Only myometrial preparations developed spontaneous contractile activity, which was effectively blocked by the calcium channel blocker nitrendipine (NTD) 10(-7) M. Pretreatment with calcium-depleted medium for 30 min. almost abolished myometrial responses to high K+ (124 mmol), oxytocin (OX) and prostaglandin F2 alpha (PGF2 alpha). Vascular responses to high K+ (124 mmol) were also nearly abolished by such treatment. However, noradrenaline (NA), vasopressin (VP) and PGF2 alpha in myometrial arteries and PGF2 alpha in chorionic vessels and stem villous arteries induced significant, but reduced contractions after calcium depletion. In all vascular preparations, exposed to calcium-depleted medium, NTD (10(-8) M) almost abolished contractions induced by calcium (0.1-4.0 mM) in the presence of K+ (124 mmol) and depressed responses to calcium in the presence of the other agonists tested. NTD (10(-10)-10(-7) M) depressed myometrial contractions induced by K+, OX and PGF2 alpha more effective than vascular responses to K+, NA, VP and PGF2 alpha in the myometrial arteries and K+ and PGF2 alpha in the placental arteries. It is concluded that activation of contraction in vessels from the human utero-placental unit implies multiple cellular sources of calcium, while in myometrial smooth muscle, influx of superficially bound calcium may be an important initial step in contractile activation. Treatment with calcium channel blockers during late human pregnancy might involve relaxation of the myometrium together with vasodilatation of the myometrial and foetal placental vascular beds.     

Relaxant effects of nifedipine on isolated, human myometrium.

Myometrial tissue was obtained from non-pregnant women subjected to hysterectomy because of various gynaecological disorders, and from women undergoing caesarean section. Strip preparations were dissected and isometric tension was recorded. Nifedipine (2.9 X 10(-8)--2.9 X 10(-6)M) inhibited spontaneous contractile activity, mainly by reducing the amplitude of contraction in both non-pregnant and pregnant myometrium. The drug also inhibited potassium induced contractions in a concentration dependent manner. This effect seemed to be more pronounced in pregnant than in non-pregnant tissue. In preparations of pregnant human myometrium, normally polarized or potassium depolarized, oxytocin induced a contractile activity that was effectively inhibited by nifedipine. Nifedipine also relaxed contractions induced by vasopressin in isolated non-pregnant myometrium. It is concluded that the relaxant effect of nifedipine on isolated pregnant and non-pregnant human myometrium can be explained by inhibition of calcium influx. The results thus support the view, that calcium influx is an important step in the initiation of contractile activity in human uterine smooth muscle.     

Involvement of calcium channels in the contractile activity of neurotensin but not acetylcholine: studies with calcium channel blockers and Bay K 8644 on the rat fundus.

The contractile activity of neurotensin and acetylcholine on rat isolated fundus strips was examined in preparations maintained in Tyrode buffer containing 2.5, 1.0 or 0 mM Ca2+. While the neurotensin contractions depended markedly on the external Ca2+ concentration, the acetylcholine-induced muscular responses were not significantly affected by omission of calcium in the superfusion media. Pre-incubation of rat fundus strips with nifedipine (0.03-3.8 microM), diltiazem (0.5-3.5 microM) or methoxyverapamil (0.3-1.3 microM) antagonized in a non-surmountable fashion the contractile activity of neurotensin but not of acetylcholine. Pretreatment with Bay K 8644 potentiated in a concentration-dependent fashion the contractile activity of rat fundus strips to neurotensin without modifying to any significant degree the acetylcholine-induced contractions. Nifedipine blocked in a concentration-dependent manner the Bay K 8644-induced potentiation of the neurotensin contractile responses in the fundus. Results demonstrate the dependence on external calcium of the contractile activity of neurotensin and the resistance of the muscarinic response to external calcium manipulations. The coupling of the neurotensin receptor to calcium channels is discussed.     

Effects of inorganic cations on K(+)-, 5-hydroxytryptamine- and noradrenaline-induced contractions of the isolated rat jugular vein and aorta

We investigated the inhibitory effects of 1 mM of the inorganic cations, La3+, Cd2+, Mn2+, Ni2+ and Co2+ on contractions induced by K+ (100 mM) and 5-hydroxytryptamine (5-HT, 10(-5) M) in the isolated rat jugular vein and on contractions induced by K+ (100 mM), 5-HT (10(-5] and noradrenaline (NA, 10(-5) M) in the rat aorta. In the venous preparation, both K(+)- and 5-HT-induced Ca2+ influx could be suppressed completely by all cations studied. In the aorta, the K(+)-induced Ca2+ influx was antagonized completely only by Cd2+. The other cations were less effective and Mn2+ was completely ineffective. Similarly to that in the venous preparation, the 5-HT-induced Ca2+ influx in the aorta was abolished by all the cations. A similar picture was found for the NA-induced Ca2+ influx with the exception of Mn2+, which antagonized the NA-induced contractions only slightly. Although organic calcium entry blockers (CEBs) (nifedipine, diltiazem, flunarizine and gallopamil) inhibited NA-induced contractions to the same extent as did Mn2+, a combination of organic CEBs and Mn2+ abolished the NA-induced Ca2+ influx completely. Apparently, organic CEBs and Mn2+ block different components of the NA-induced Ca2+ influx pathway.     

Effects of yohimbine, rauwolscine and corynanthine on contractions and calcium fluxes induced by depolarization and prostaglandin F2 alpha in rat aorta

The effects of the selective alpha 2-adrenoceptor antagonists yohimbine and its stereo-isomer rauwolscine and the selective alpha 1-adrenoceptor antagonist corynanthine (a third yohimbine stereoisomer) on contractions induced in rat aorta by depolarization and prostaglandin F2 alpha (PGF2 alpha) have been compared. In calcium-free solution, depolarization with 100 mM K+ failed to produce a contraction of rat aorta but PGF2 alpha (3 microM) stimulated a contraction equal to about 23% of maximal elicited in normal physiological solution. Yohimbine had no significant effect on depolarization-induced contractions except at concentrations greater than 30 microM. Rauwolscine and corynanthine (1 to 100 microM) depressed depolarization-induced contractions in a concentration-dependent manner, but the characteristics of inhibition were not identical. Contractions induced by PGF2 alpha (3 microM) were depressed in a concentration-dependent manner by rauwolscine (3 to 100 microM) but were unaffected by yohimbine or corynanthine. Depolarization-stimulated 45Ca influx was depressed by rauwolscine and corynanthine to about the same extent as were the contractions; while rauwolscine (100 microM) completely inhibited PGF2 alpha-stimulated 45Ca influx, it also depressed part of the PGF2 alpha-stimulated contraction dependent on intracellular calcium. Rauwolscine (100 microM) partly inhibited PGF2 alpha-stimulated release of 45Ca from aortic smooth muscle in calcium-free solution. It is concluded that the yohimbine structure possesses a calcium entry blocking action as well as a depressant action on contractions not dependent on calcium entry. The predominant effect depends on the structural configuration and the nature of the stimulating agent.     

Inhibitory effect of noradrenaline uptake inhibitors on contractions of rat aortic smooth muscle.

1 The effects of noradrenaline (NA) uptake inhibitors on contractions induced by NA, high K+, and 12-O-tetradecanoylphorbol-13-acetate (TPA) in rat isolated aorta were investigated. 2 Protriptyline (0.3 microM) and amitriptyline (0.3 microM) produced an approximately parallel shift to the right in the dose-response curve to NA. Protriptyline (> 0.3 microM), amitriptyline (> 0.3 microM) and xylamine (0.01-1 microM) significantly reduced the maximal contractile response to NA. The IC50 values for inhibition of the contractile response to 3 microM NA were 1.58 microM for xylamine, 1.70 microM for amitriptyline and 2.57 microM for protriptyline. 3 Protriptyline and amitriptyline dose-dependently inhibited the high K+ (60 mM)-induced contraction (IC50 = 0.69 microM for protriptyline and IC50 = 3.15 microM for amitriptyline). In contrast, xylamine did not affect the high K(+)-induced contraction. 4 Protriptyline and amitriptyline dose-dependently inhibited TPA (1 microM)-induced contraction in calcium-free solution; xylamine (up to 30 microM) was without effect. Staurosporine (10 nM) completely inhibited the TPA- and NA-induced contraction. 5. Protriptyline (3 microM) and amitriptyline (3 microM) caused about 54% and 60% inhibition, respectively, of aortic contractions caused by endothelin-1 (10 nM) in the absence of endothelium. Xylamine (10 microM) was without effect. 6 Inhibitory effects of NA uptake inhibitors on contractions were independent of the presence of endothelium and were unaffected by the K+ channel blockers, tetraethylammonium ions (up to 3 mM) and glibenclamide (up to 30 microM). 7 These results indicate that tricyclic antidepressant drugs such as protriptyline and amitriptyline could act as both postsynaptic adrenoceptor antagonists and direct inhibitors of muscle contraction; whereas, xylamine, a structurally distinct NA uptake blocker might principally exert its action only at alpha-adrenoceptors on rat aortic smooth muscle.     

Excitatory effect of Clostridium perfringens alpha toxin on the rat isolated aorta.

Clostridium perfringens alpha toxin caused contraction of the isolated aorta of the rat in a dose-dependent manner. The contractile action caused by the toxin was inhibited or abolished by calcium antagonists such as nifedipine, verapamil and cinnarizine, or a Ca-free medium, but was not affected by phentolamine, chlorpheniramine, atropine, tetrodotoxin or a low Na medium. The toxin stimulated Ca uptake into the aorta in a dose-dependent manner. 8-N,N'-diethylaminooctyl-3,4,5-trimethoxybenzoate (TMB-8) blocked significantly both the toxin- and noradrenaline (NA)-induced contractions. Trifluoperazine (TFP) and N-(6-aminohexyl)-5-chloro-1-naphtharene sulphonamide (W-7) did not affect the contractile activity of the toxin but blocked the NA-induced contraction. The toxin also stimulated the 32P phosphate labelling of phosphatidylinositol (PI) and phosphatidic acid (PA) in the preparation. These results indicate that the toxin-induced contraction, which is different from that induced by NA, is the result of a direct action of the toxin on the aorta and is due to an increased Ca2+ permeability across the smooth muscle membrane. It is suggested that the contractile response to the toxin is associated with activation of phospholipid metabolism and enhanced entry of Ca into the aorta.     

Multiple actions of glaucine on cyclic nucleotide phosphodiesterases, alpha 1-adrenoceptor and benzothiazepine binding site at the calcium channel.

1. In the present study, the properties of glaucine (an aporphine structurally related to papaverine) were compared with those of papaverine, diltiazem, nifedipine and prazosin. The work includes functional studies on rat isolated aorta contracted with noradrenaline, caffeine or KCl, and a determination of the affinity of glaucine at calcium channel binding sites of alpha-adrenoceptors, by use of [3H]-(+)-cis-diltiazem, [3H]-nitrendipine and [3H]-prazosin binding to cerebral cortical membranes. The effects of glaucine on the different molecular forms of cyclic nucleotide phosphodiesterases (PDE) isolated from bovine aorta were also determined. 2. Contraction evoked by noradrenaline (1 microM) or depolarizing solution (60 mM KCl) were inhibited in a concentration-dependent manner by all the compounds tested. As expected, prazosin showed a greater selectivity of action on NA-induced contraction, whereas nifedipine and diltiazem appeared more potent on KCl-induced contraction. Glaucine had a greater potency on the contraction elicited by noradrenaline whereas papaverine acted non specifically. 3. In Ca(2+)-free solution, prazosin (0.1 microM) and glaucine (0.1 mM) inhibited the contraction evoked by NA; diltiazem (0.1 mM) diminished this contraction whereas nifedipine (1 microM) had no effect. Preincubation of tissues with glaucine, diltiazem, nifedipine and prazosin did not modify the contractile response induced by caffeine. In contrast, papaverine (0.1 mM) significantly inhibited the contractions evoked by NA or caffeine in Ca(2+)-free medium. 4. Glaucine and papaverine show affinity at the [3H]-prazosin binding site and at the benzothiazepine binding site of the Ca(2+)-channel receptor complex, but have no effect at the dihydropyridine binding site in rat cerebral cortex.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of digoxin in isolated human pulmonary vessels.

In isoalted human pulmonary arteries and veins the contractile response to noradrenaline (1.8 X 10(-5)M) was 33 +/- 7.4% and 20 +/- 4.5% (Mean +/- S.E.M.) of that induced by potassium (127 mM). A variable degree of spontaneous contractile activity was recorded in the vein preparations. This activity was abolished by nifedipine (2.9 X 10(-6)M). Digoxin (10(-6)M) induced contractions in pulmonary vessels. In the arteries, the digoxin contraction developed slowly and reached a maximum amplitude of 90 +/- 4% (Mean +/- S.E.M.) of the potassium evoked contraction. In the veins, the amplitude of the digoxin contraction was 32 +/- 5% of that induced by potassium. Digoxin (10(-6)M) also increased the maximum response to noradrenaline and potassium in both arteries and veins. In the arteries, the noradrenaline and potassium contractions increased to 211 +/- 6.8% and 145 +/- 8.9 of control, and in the veins to 169 +/- 13.5% and 163 +/- 9.9%, respectively. Nifedipine in concentrations which completely relaxes arterial and venous preparations contracted by potassium, had only a slight relaxing effect on digoxin induced contraction. It is concluded that digoxin increases the tension in pulmonary arteries and veins, and increases the maximum response to noradrenaline and potassium in both types of vessels. The digoxin induced contraction is highly resistant to blockade of extracellular calcium influx.