Comparative effects of verapamil and sodium nitroprusside on contraction and 45Ca uptake in the smooth muscle of rabbit aorta, rat aorta and guinea-pig taenia coli.

The effects of verapamil and sodium nitroprusside on muscle tension and 45Ca uptake activated in different ways were compared in rabbit aorta, rat aorta and guinea-pig taenia coli. In rabbit aorta, K-induced contraction was specifically inhibited by verapamil and noradrenaline-induced contraction by sodium nitroprusside. In rat aorta, both K-induced and noradrenaline-induced contractions were inhibited by verapamil or by sodium nitroprusside also. In taenia, both K- and histamine-induced sustained contractions were inhibited by verapamil but not by sodium nitroprusside. The effect of verapamil was competitively antagonized by external Ca, while that of sodium nitroprusside was not. High K, noradrenaline and histamine increased the rate of 45Ca uptake in aortae and taenia. In rabbit aorta the increment in response to high K was specifically inhibited by verapamil and the increment induced by noradrenaline was specifically inhibited by sodium nitroprusside. In rat aorta, increments induced by both high K and noradrenaline were inhibited by verapamil and by sodium nitroprusside. In taenia, the increments induced by high K and by histamine were inhibited by verapamil but not by sodium nitroprusside. These results suggest different characteristics of Ca entry systems in these smooth muscles. In rabbit aorta, there seem to be two Ca channels, one of which is activated by high K and inhibited by verapamil, while the other is activated by noradrenaline and inhibited by sodium nitroprusside. In rat aorta, both K- and noradrenaline-activated Ca pathways are sensitive to both verapamil and sodium nitroprusside whereas, in taenia, both K- and histamine-activated Ca pathways are sensitive only to verapamil.     

Effects of calmodulin antagonists on tension and cellular calcium content in depolarized vascular and intestinal smooth muscles.

1 Several putative calmodulin antagonists have been examined for their inhibitory action on muscle tension and cellular Ca content in the K-depolarized vascular and intestinal smooth muscles. 2 The 65.4 mM K-induced sustained contraction in the media-intimal layer of rabbit aorta and the 45.4 mM K-induced sustained contraction in guinea-pig taenia coli were inhibited by the calmodulin antagonists, prenylamine, chlorpromazine, N2-dansyl-L-arginine-4-t-butylpiperadine amide (No. 233), and N-(6-aminohexyl)-5-chloro-1-naphthalenesulphonamide (W-7), and also by the organic Ca antagonists, verapamil and diltiazem. 3 The cellular Ca content in rabbit aorta and guinea-pig taenia coli as measured by a modified lanthanum technique increased in the high-K solutions. The increments were inhibited by these antagonists at concentrations similar to those required to inhibit the K-induced contractions. However, W-7 did not change (in aorta) or only slightly decreased (in taenia coli) the K-induced increase in the cellular Ca content. 4 A high concentration (2 X 10(-4)M) of W-7 increased the resting cellular Ca content without increasing the muscle tension in aorta. The increment was inhibited by verapamil, sodium nitroprusside or hypoxia (N2 aeration). 5 It is suggested that the inhibitory effects of prenylamine, chlorpromazine and No. 233 may be attributed mainly to the Ca antagonistic effect whereas W-7 may inhibit the process beyond the transmembrane Ca influx.     

The inhibitory effects of N2-dansyl-L-arginine-4-t-butylpiperidine amide (TI233) on contraction of vascular and intestinal smooth muscle.

Effects of N2-dansyl-L-arginine-4-t-butylpiperidine amide (TI233) on the contractions in vascular and intestinal smooth muscles were examined. High K-induced sustained contractions in the smooth muscles were inhibited by TI233 with an IC50 of 2.1 X 10(-5) M for rabbit aorta and 3.6 X 10(-6) M for guinea-pig taenia coli in a solution containing 1.5 mM Ca. Initial transient contraction induced by K in taenia coli was less sensitive to the inhibitory effect of TI233. When the Ca concentration in the medium was decreased to 0.3 mM, the concentration-inhibition curves for TI233 shifted to the left in both aorta and taenia coli. Increasing the Ca concentration to 7.5 mM shifted the curve to the right in the aorta. TI233 also inhibited the noradrenaline-induced contraction in the aorta (IC50 = 2.1 X 10(-5 M). In a hypoxic solution without added glucose, the inhibitory effect of TI233 on the K-induced contraction in aorta was augmented. In the presence of high concentrations (40 mM) of glucose in hypoxia, TI233 did not inhibit the noradrenaline-induced contraction of the aorta. Hypoxia and a high concentration of glucose also decreased the inhibitory effect of TI233 on the K-induced contraction in taenia coli. TI233 inhibited the K-induced increase in cellular Ca content measured by a modified lanthanum method. TI233 decreased oxygen consumption and ATP content of resting and K-stimulated aorta and taenia coli. It was concluded that TI233 inhibits the vascular and intestinal smooth muscle contraction by a Ca antagonistic action and also by inhibition of aerobic metabolism.     

Effects of the calcium channel facilitator, CGP 28,392, on different modes of contraction in smooth muscle of rabbit and rat aortae and guinea-pig taenia caeci.

Effects of a Ca2+ channel facilitator, CGP 28,392, on smooth muscle contractions were examined in order to delineate characteristics of Ca2+ channels in rabbit and rat aortae and guinea-pig taenia caeci. Application of increasing concentrations of KCl induced contractile responses in these smooth muscles and CGP 28,392 shifted the concentration-response curve for KCl to the left. The maximum response was also increased in rat aorta and guinea-pig taenia. CGP 28,392 also shifted the concentration-response curves for noradrenaline in rat aorta and for histamine in taenia to the left and increased the maximum response in rat aorta. However, the corresponding curve for noradrenaline in rabbit aorta was not affected by CGP 28,392. The sustained contractions induced by KCl were inhibited by cumulative application of verapamil in these smooth muscles. Pretreatment of the muscle with CGP 28,392 decreased the inhibitory effect of verapamil. The noradrenaline-induced contraction in rat aorta and the histamine-induced contraction in taenia were also inhibited by verapamil, and CGP 28,392 antagonized the effect of verapamil. The noradrenaline-induced contraction in rabbit aorta was only slightly inhibited by verapamil, and CGP 28,392 did not modify the effect of verapamil. In these smooth muscles, cumulative application of Ca2+ to the Ca2+-depleted, KCl-treated muscle induced contraction, and the concentration-response curve for Ca2+ was shifted to the left by CGP 28,392 and to the right by verapamil. The concentration-response curves for Ca2+ in Ca2+-depleted, noradrenaline-treated rabbit and rat aortae and in Ca2+-depleted, histamine-treated taenia were also shifted to the left by CGP 28,392 and to the right by verapamil.(ABSTRACT TRUNCATED AT 250 WORDS)     

The inhibitory effect of X537A on vascular and intestinal smooth muscle contraction

Summary the effects of X537A (Lasalocid) on contractions induced in vascular and intestinal smooth muscles were examined. High K-induced sustained contractions were inhibited by X537A with an IC50 of 2.8·10^–6 M in rabbit aorta and 8.8·10^–8 M in guinea-pig Taenia coli. Changing the Ca concentration in the medium did not modify the effect of X537A. X537A also inhibited the noradrenaline-induced contraction in aorta (IC50=7.0·10^–8 M). In a solution without added glucose, the inhibitory effect of X537A on the K-induced contraction in aorta was augmented (IC50=1.5·10^–8 M). Under hypoxic conditions, the inhibitory effects of X537A on the noradrenaline-induced contractions in aorta and on the K-induced contraction in T. coli were decreased (IC50>10^–5 M and 3.2·10^–6 M, respectively). X537A inhibited the K-induced increase in cellular ⁴⁵Ca content in aorta measured by a modified lanthanum method. However, the IC50 for ⁴⁵Ca uptake (6.6·10^–7 M) was lower than that for K-induced contraction. In both tissues, X537A decreased the ATP content. Oxygen consumption of rat liver mitochondria was inhibited by X537A. From these results, it is concluded that the inhibitory effect of X537A on the noradrenaline-induced contraction in aorta and the K-induced contraction in T. coli, but not the K-induced contractions in aorta, may be due to the inhibition of aerobic metabolism.     

SODIUM-DEPENDENT RELAXATION IN ARTERIAL SMOOTH MUSCLE *

1. The effects of isosmotic substitution of choline for sodium on resting tension and on relaxation after noradrenaline-induced contraction was studied in rabbit isolated aortic strips immersed in Hepes-buffered physiologic salt solution (PSS) warmed to 37°C and gassed with 100% O₂. 2. Isosmotic substitution of choline for sodium produced a sustained increase in resting tension which effectively prevented any evaluation of the influence of sodium on relaxation. The increase in resting tension was insensitive to 10^?8 mol/1 atropine but was abolished by 10 min exposure to calcium-free PSS prior to replacement of sodium. 3. Under sodium-calcium free conditions which eliminated the increase in resting tension observed in sodium-free PSS, stimulation with 10^?5 mol/1 noradrenaline initiated contractions that were 55 ± 7.5% of the control response in normal PSS. Washout of noradrenaline with sodium-calcium-free PSS failed to produce any decrement in tension. However, restoration of the normal sodium resulted in gradual relaxation. 4. These results suggest that sodium is required for relaxation after noradrenaline-induced contraction of arterial smooth muscle.     

Species differences in the inhibitory effect of ouabain on high K-induced contraction in the ileal longitudinal muscle

Effects of ouabain on a high K-induced contraction and intracellular Na and K contents of the ileal longitudinal muscle preparations isolated from various animal species was compared. In muscles of various animals (monkey, dog, cat, rabbit, guinea-pig, rat and mouse), the high K-induced contraction was inhibited by verapamil (5 X 10(-8) M) or ouabain (3 X 10(-6) M), and both the inhibitions were antagonized by an increase in external Ca in a competitive manner. Species differences were shown in the ouabain effect but not in the verapamil one. Regarding the sensitivity to ouabain, the muscles were divided into two groups, that is, the muscle from monkey, rabbit, guinea-pig or dog belongs to the high sensitivity group, and that from cat, rat or mouse belongs to the low one. The order of sensitivities of the muscles to ouabain in the relaxation was consistent with that in Na,K-ATPase inhibition, as reported by Repke et al. (1965). Intracellular Na contents of muscles were increased by an addition of ouabain to the high K solution, and the rate and amount of the accumulation of intracellular Na varied in these muscles. Except for the cat muscle, a high correlation was noted between the ouabain-induced relaxation and the amount of intracellular Na accumulation. However, the regression coefficients were variable: 4.1 in the monkey muscle, 2.2 in the rat one, and about 1.0 in the others. That is, the monkey muscle showing the high regression coefficient value probably has a high sensitivity to the inhibition of intracellular Na in the high K-induced contraction.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of sodium nitroprusside on cytosolic calcium level in vascular smooth muscle

The inhibitory effect of sodium nitroprusside on the cytosolic Ca2+ level ([Ca2+]cyt), measured simultaneously with contraction by means of a fluorescence dye, fura-2, and on the 45Ca2+ uptake was tested in the isolated rat aortic smooth muscle. Norepinephrine increased muscle tension, 45Ca2+ uptake and [Ca2+]cyt. In a Ca2+-deficient solution, norepinephrine transiently increased muscle tension and [Ca2+]cyt. Sodium nitroprusside inhibited all changes induced by norepinephrine although the inhibition of [Ca2+]cyt was less than that of muscle contraction. Sodium nitroprusside also inhibited the high K+-induced contraction at concentrations higher than those needed to inhibit norepinephrine-induced contraction. Inhibition of the high K+induced contraction was accompanied by a small decrease in [Ca2+]cyt and a smaller decrease in 45Ca2+ uptake. Methylene blue antagonized, and M&B 22,948 potentiated the inhibitory effect of sodium nitroprusside. These results suggest that sodium nitroprusside has multiple sites of action. At relatively low concentrations, sodium nitroprusside could inhibit the Ca2+ influx and Ca2+ release. At relatively high concentrations, this inhibitor could also augment Ca2+ sequestration and decrease the sensitivity of contractile elements to Ca2+.     

DIFFERENCES IN ENDOTHELIUM-DEPENDENT RELAXATION IN VARIOUS ARTERIES FROM WATANABE HERITABLE HYPERLIPIDAEMIC RABBITS WITH INCREASING AGE

1. Endothelium-dependent relaxation in response to acetylcholine (ACh) and the calcium ionophore A 23187 was examined in aorta, coronary, basilar and renal arteries isolated from Watanabe heritable hyperlipidaemic (WHHL) rabbits of 2, 6 and 12 months of age, with normolipidaemic heterozygous WHHL rabbits as controls. 2. In the rings of WHHL rabbit aortae and coronary arteries preconstricted with vasoconstrictors, endothelium-dependent relaxation in response to ACh was attenuated with age compared to the heterozygous WHHL rabbits. A significant negative correlation was found between the total cholesterol content and the relaxation response to ACh in the aortae or coronary arteries from 6 and 12 month old WHHL rabbits. 3. In the rings of basilar arteries, endothelium-dependent relaxations to ACh were not modified with age. Similarly, in the rings of renal arteries, the relaxation response to ACh was not changed with age, but in the 6 and 12 month preparations, after the age of 6 months, a contraction following the relaxation appeared at higher concentrations of ACh (10^?7 to 10^?6 mol/L). The contraction was endothelium-dependent and inhibited by indomethacin. 4. A 23187-induced endothelium-dependent relaxations were also markedly attenuated in the aorta and significantly in the coronary artery with age. 5. Endothelium-independent relaxation to sodium nitroprusside was not changed in all arteries from WHHL rabbits of different ages. 6. These findings indicate that in the aorta and coronary artery of the WHHL rabbit, the endothelium-dependent relaxation to ACh and A 23187 becomes impaired with increasing age (i.e., with the progression of cholesterol deposition in the arterial wall) but is preserved in the basilar and renal artery.     

Effects of nifedipine derivatives on smooth muscle cells and neuromuscular transmission in the rabbit mesenteric artery

The effects of nifedipine and its derivatives, nisoldipine, nimodipine and nitrendipine, on smooth muscle cells and neuromuscular transmission were investigated in the rabbit mesenteric artery. These agents in concentrations of up to 2 X 10(-7) M modified neither the membrane potential nor the membrane resistance, yet did inhibit the spike potential evoked by direct muscle stimulation in the presence of TTX or by perivascular nerve stimulation. The inhibitory action of nitrendipine was weaker than that of the other derivatives. These agents had no effect on the miniature excitatory junction potentials (m.e.j.ps) and e.j.ps evoked by the first stimuli and after completion of the facilitation in a train stimulation. Nifedipine and its derivatives had no effect on the K-induced depolarization but did have a marked effect on the K-induced contraction. Nisoldipine showed the highest inhibitory potency for the K-induced contraction [IC50 was 1.2 X 10(-9) M for the 128 mM (K)o-induced contraction]. Noradrenaline depolarized the membrane (greater than 5 X 10(-7 M) and produced contraction (greater than 3 X 10(-7) M). The contraction evoked by high concentrations of noradrenaline was inhibited by these agents to a greater extent than that evoked by low concentrations. The contraction evoked by perivascular nerve or direct muscle stimulation was partly inhibited by nifedipine and its derivatives. The contraction elicited by Na-free solution was inhibited by these agents but the noradrenaline- or caffeine-induced contraction in Ca-free solution was not. These results indicate that in smooth muscle cells of the rabbit mesenteric artery, nifedipine and its derivatives inhibit the voltage dependent Ca-influx which occurs during the spike potential and in response to K-, electrically- or noradrenaline-induced depolarization. These derivatives appear to have no effect on the adrenoceptor operated Ca increase in myoplasm which occurs in the absence of depolarization at low noradrenaline concentrations. The derivatives act as Ca antagonists with a quantitative difference in potency, i.e. the strongest action was observed with nisoldipine and the weakest with nitrendipine.     

Nitroglycerine-induced biphasic relaxation in vascular smooth muscle of rat aorta

Nitroglycerine induced biphasic relaxation in the rat aorta, previously contracted by noradrenaline; a rapid decrease in tension was followed by a gradual increase reaching a steady level below the control contractile tension. No initial transient relaxation was induced by nitroglycerine in high K-stimulated muscle. The initial transient relaxation, but not the sustained relaxation, was dependent on the concentration of external K; maximum relaxation was observed in the presence of 2.7 mM K solution and only a slight relaxation was observed in 0 mM or 10.8 mM K solution. The initial transient relaxation was also inhibited by ouabain or low Na solution. On an appropriate increase in the concentration of external K, noradrenaline-induced contraction was transiently relaxed. Previous application of nitroglycerine potentiated this K-induced relaxation. Pretreatment of the muscle with methylene blue, an inhibitor of guanylate cyclase, inhibited both the initial transient and the sustained relaxations induced by nitroglycerine, but not the K-induced transient relaxation. It is suggested that the nitroglycerine-induced initial transient relaxation, but not the sustained relaxation, may be due to a stimulation of an electrogenic Na pump. Both relaxation phases may be mediated by cyclic GMP.     

心房肽Ⅲ对人体血管作用的差异性

采用离体人脐动脉,人胃网膜右动脉,家兔及大白鼠胸主动脉肌条,观察APⅢ和NP的作用。APⅢ75nmol/l对由NA3μmol/l,5-HT1μmol/l,HT10μmol/l,PE 1μmol/l收缩的家兔及大白鼠胸主动脉肌条具有明显松弛作用,对由上述药物及KCl 80mmol/l,PGF_(2α)10μmol/l所收缩的人脐动脉及人胃网膜右动脉并无松弛作用。NP 1μmol/l对由NA3μmol/l,5-HT 1μmol/l,HT 10μmol/l等所收缩的两种人血管都有明显松弛作用,并能降低血管基本张力。结果提示:APⅢ对家兔及大白鼠的胸主动脉有松弛作用,但对人脐动脉及人胃网膜右动脉无松弛作用,表明APⅢ对血管的作用可能具有动物种属或血管种类差异性,NP则有较普遍的血管松弛作用。     

Contractile response of rabbit dilator pupillae muscle to isotonic high K or norepinephrine, and Ca movement

Isotonic 80 mM K and norepinephrine contracted the isolated dilator pupillae muscle of the rabbit. Norepinephrine-induced contraction was inhibited by papaverine, but not by Ca blockers, while K-induced contraction was abolished by removing Ca from the bathing fluid but there was a residual response to norepinephrine in Ca-free solution. These results suggest that, in the rabbit dilator pupillae muscle, K-induced contraction is mainly attributable to the increase of Ca influx, while norepinephrine-induced contraction is mainly due to the facilitation of release of intracellularly sequestered Ca.     

用兔主动脉条研究小蘖碱和硝普钠对几种激动剂作用的影响

用离体家兔主动脉条观察Ber和NP的作用;NP降低血管条的基本张力,明显逆转由PE,HT、5-HT和KCl所引起的收缩,Ber仅逆转PE引起的收缩,对HT,5-HT和KCl引起的收缩无影响,也不影响血管条的基本张力。 Ber和Pra均阻断PE对血管条的收缩作用,两药的IC_(50)分别为30μM和0.2μM。 实验结果表明;NP非特异性逆转几种激动剂所致血管条的收缩,Ber仅逆转PE引起的血管收缩,其作用性质和Pra相似,提示Ber对血管条的α_1-肾上腺素受体具有选择性阻断作用。     

Mechanisms underlying the inhibitory effect of dibutyryl cyclic AMP in vascular smooth muscle.

The mechanism by which dibutyryl cyclic AMP (db-cAMP) induces vasodilatation was examined in isolated rat aorta. The contraction induced by norepinephrine (NE) was more sensitive to the inhibitory effect of db-cAMP than that induced by high K+, and the contraction induced by lower concentrations of each stimulant was more sensitive to db-cAMP than that induced by higher concentrations. Db-cAMP at 10 microM inhibited the increases in muscle tension and cytosolic Ca2+ level ([Ca2+]i) without changing the [Ca2+]i-tension relationship, suggesting that the inhibitory effect is mainly due to a decrease in [Ca2+]i. A higher concentration (300 microM) of db-cAMP inhibited muscle tension more strongly than [Ca2+]i suggesting that db-cAMP decreases Ca2+ sensitivity of contractile elements. In contrast, 10 microM verapamil inhibited the NE-stimulated [Ca2+]i more strongly than the NE-induced contraction. The verapamil-insensitive portion of the NE-stimulated [Ca2+]i and contraction was inhibited by db-cAMP, suggesting that db-cAMP and verapamil act by different mechanisms. In Ca(2+)-free solution, 1 microM NE induced transient increases in muscle tension and [Ca2+]i. The transient contraction was inhibited by 1 mM db-cAMP more strongly than [Ca2+]i. An activator of adenylate cyclase, forskolin, showed inhibitory effects similar to those of db-cAMP. The inhibitory effects of db-cAMP and forskolin were inversely proportional to [Ca2+]i before the addition of these inhibitors. These results suggest that db-cAMP inhibits smooth muscle contraction by decreasing [Ca2+]i and the Ca2+ sensitivity of contractile elements, and that both of these effects are stronger when [Ca2+]i is lower.     

The effects of diltiazem (CRD-401) on the membrane and mechanical properties of vascular smooth muscles of the rabbit.

1 The effects of diltiazem on electrical and mechanical properties of vascular smooth muscles of the rabbit were examined by various experimental procedures. 2 In the pulmonary artery, diltiazem (0.1 to 10 microgram/ml) did not modify the membrane potential (-56 mV), length constant of the tissue (1.47 mm) or rectifying properties of the membrane. Diltiazem (0.1 to 10 microgram/ml) did not modify the membrane potential of the mesenteric artery (-62.5 mV). 3 Diltiazem (1 to 10 microgram/ml) suppressed mechanical responses of pulmonary and mesenteric arteries induced either by direct stimulation of the muscle (1.0 s pulse) or by neural activation (0.5 ms pulse, 30 Hz and 10 s total duration). Diltiazem suppressed the contraction induced by nerve stimulation to a greater extent than that induced by direct muscle stimulation. 4 When the depolarization-contraction relationship of the smooth muscle of the pulmonary artery was observed by voltage clamp technique, diltiazem (1 to 10 microgram/ml) raised the critical membrane potential to evoke contraction from 5 mV to 12 mV, and reduced the amplitude of contraction obtained at any given depolarization level. 5 In the pulmonary artery, diltiazem (10 microgram/ml) suppressed K-induced contraction and raised the mechanical threshold, while K-induced depolarization was not suppressed. Diltiazem (1 to 10 microgram/ml) also suppressed noradrenaline-induced contraction, raised the mechanical threshold and suppressed noradrenaline-induced depolarization. 6 The vasodilator actions of diltiazem on the vascular smooth muscle were compared to vasodilator actions observed with other Ca-antagonists.     

Role of Ca2+ in non-cholinergic,non-adrenergic responses to nerve stimulation of the guinea-pig small intestine

Summary The effects of calcium channel blockers (D-600, verapamil), sodium nitroprusside, papaverine, indomethacin, local anaesthetics and blockade of sodium pump activity on the non-cholinergic, non-adrenergic transmission in the guinea-pig duodenum, jejunum, proximal and terminal ileum were analysed in the presence of atropine and guanethidine.A decrease of the extracellular Ca²⁺ concentration inhibited the primary and rebound contractions but only in Ca²⁺-free solution was the primary relaxation diminished. D-600, verapamil, sodium nitroprusside and papaverine inhibited both the primary and rebound contractions to the same degree and their effects on the primary relaxation were less pronounced than on the contractions.Indomethacin dissolved in alkaline solution did not depress the non-cholinergic, non-adrenergic responses in any region of the small intestine, whereas indomethacin dissolved in ethanol antagonized both the primary and rebound contractions in the muscles.Local anaesthetics (procaine, trimecaine) in low concentrations inhibited only the primary contraction. Higher concentrations also inhibited both the rebound contraction and primary relaxation. Procaine in low concentrations did not markedly affect the non-cholinergic, non-adrenergic i.j.p.s and e.j.p.s., but did block the action potentials induced by e.j.p.s.Our findings indicate that the primary relaxation, and primary and rebound contractions are probably induced by different mechanisms and are not mediated by ATP. We confirmed that prostaglandins did not participate in the generation of the rebound contraction.This study was supported in part by JSPS Japan     

Effects of basal and acetylcholine-induced release of endothelium-derived relaxing factor on contraction to alpha-adrenoceptor agonists in a rabbit artery and corresponding veins.

1. The effects of an endothelium-dependent (acetylcholine) and an endothelium-independent (sodium nitroprusside) relaxant against noradrenaline-induced contractions were compared in three isolated superficial blood vessels of the rabbit, the lateral saphenous vein, plantaris vein and distal saphenous artery. Both produced concentration-related relaxations of all three vessels and were more effective against submaximal than maximal contractions to noradrenaline. Transient contractions to high concentration of acetylcholine occurred only in endothelium-intact preparations of saphenous vein and were inhibited by flurbiprofen. 2. In endothelium-denuded preparations sodium nitroprusside was 3 times more effective than in endothelium-intact preparations, while acetylcholine (less than 3 microM) was inactive. Sensitivity was similar for each relaxant: lateral saphenous vein greater than or equal to plantaris vein greater than distal saphenous artery. The similar profile of sodium nitroprusside and acetylcholine suggests that differences in susceptibility to endothelium-derived relaxing factor (EDRF) are caused by inter-vessel variations in the excitation-coupling process for noradrenaline. 3. Haemoglobin inhibited acetylcholine-induced relaxations in the endothelium-intact preparation of the lateral saphenous vein and distal saphenous artery, which suggests a similar EDRF in each preparation and the likelihood that this is a single substance, presumably nitric oxide. 4. The influence of basal, spontaneously released EDRF on alpha-adrenoceptor function was tested either by mechanical disruption of the endothelium or by adding haemoglobin to endothelium-intact segments. Endothelial disruption slightly reduced contractions to noradrenaline (NA) in distal saphenous artery but increased response size of lateral saphenous and plantaris veins, in the latter also increasing sensitivity to NA: haemoglobin mimicked endothelial disruption. Thus, basal release of EDRF like acetylcholine and nitroprusside was more effective in the veins than in the corresponding artery. 5. In lateral saphenous vein responses to phenylephrine were enhanced by endothelial disruption, but without change in sensitivity: responses to UK-14304, B-HT 920 and cirazoline, which had a relatively slow speed of onset of contraction were not affected. There was no correlation between enhancement and alpha-adrenoceptor sub-type although the agonists which were enhanced all activate alpha 1-adrenoceptors. Competitive antagonists failed to reveal an alpha-adrenoceptor subtype enhanced by endothelial disruption. However, effects of phenoxybenzamine suggest that alpha 1-adrenoceptors are necessary for the influence of basal EDRF.     

Developmental aspects of bladder contractile function: sensitivity to extracellular calcium.

The urinary bladders of 1-day and 1-week-old rabbits generate higher intravesical pressures in response to bethanechol and field stimulation than bladders isolated from mature 8-week-old rabbits. Yet the density of cholinergic receptors in the rabbit bladder does not change with maturation (1 day to 8 weeks). In an effort to better understand the molecular mechanisms by which newborn rabbit bladders generate greater pressures than the bladders of adult rabbits, we studied the effect of maturation on the relationship between extracellular calcium and contraction. Our results showed quite clearly that (1) at physiologic concentrations of calcium (2.5 mumol/l), isolated bladder strips of 1-day- and 1-week-old rabbits contracted in response to bethanechol to 98% of their maximal tension as opposed to 68% for their 8-week-old counterparts, (2) the ED50 (for calcium) for the 1-day and 1-week bladders was 0.4 mmol/l whereas the ED50 for the adult bladder strips was 2.2 mmol/l, and (3) the neonatal bladders demonstrated a much greater sensitivity to diltiazem than the adult bladders. The contractile response to calcium of the neonatal bladders was significantly inhibited by 1 mumol/l diltiazem whereas the 8-week-old bladders showed no inhibition at this concentration. In a second series of experiments, the effect of extracellular calcium on concentration was correlated with the intracellular concentration of free calcium using the calcium fluoride FURA-2 and surface spectrofluorometry. These studies confirmed that the increased contractile response of the neonatal bladder strips to calcium or cholinergic agonists was associated with an increase in the maximal intracellular free calcium concentration.     

Mode of action of sodium nitroprusside on vascular smooth muscle

1. Sodium nitroprusside is a potent relaxant of smooth muscles with a predominantly tonic response, e.g. rat aorta contracted by noradrenaline, angiotensin II, Phe2-Lys8-vasopressin, BaC1(2), or KC1, and guinea-pig tracheal smooth muscle contracted by carbachol. 2. Smooth muscle preparations from the splanchnic region and with varying degrees of phasic contractility are less sensitive and develop tachyphylaxis (portal vein, duodenum of the rat) or are unresponsive to sodium nitroprusside (vas deferens, uterus of the rat). 3. Cardiac auricles of the guinea pig are not affected by sodium nitroprusside in either frequency or amplitude or spontaneous contractions. 4. Sdium nitroprusside causes a parallel shift of the dose-response curve of rat aorta to noradrenaline to the right and reduces the maximum response. 5. The drug has no blocking or stimulant effect on alpha- or beta-adrenoceptors, respectively. 6. Sodium nitroprusside inhibits the contractile response of calcium-depleted depolarized rat aorta to extra-cellular calcium. Like verapamil, it inhibits the increment in 45calcium uptake of rabbit aorta elicited by K+. Sodium nitroprusside significantly reduced 45calcium binding by microsomes prepared from rabbit aorta. 7. Rabbit aorta was incubated with lanthanum chloride to prevent calcium influx; sodium nitroprusside reduced the maintained rapid contraction phase in response to noradrenaline which is believed to be based on the intracellular activation of calcium. 8. In rat aorta, cellular cAMP and ATP levels were not found to be affected by the drug. 9. Rabbit aorta, "skinned" by glycerination is unresponsive to sdoium nitroprusside. 10. It is concluded that sodium nitropruside acts on exictation-contraction coupling predominantly in tonic smooth muscle by interfering with both the influx and the intracellular activation of calcium.