Role of nitric oxide in the contraction of circular muscle in the rat portal vein

The role of nitric oxide in the electrical and mechanical activities of the rat portal vein was examined in circular muscle preparations with intact endothelium that were isolated from the longitudinal muscle layer. In contrast to the longitudinal muscle preparation, the circular muscle preparation did not show spontaneous phasic contraction. Inhibition of nitric oxide synthesis by Nomega-nitro-L-arginine (L-NNA) induced a tonic contraction. The contraction was inhibited by L-arginine, sodium nitroprusside or nifedipine. L-NNA did not induce contraction in endothelium-damaged preparations. The membrane potential of smooth muscle cells recorded in endothelium-intact preparations showed sporadic action potentials. L-NNA increased the frequency of action potentials without changing the resting membrane potential. The action potentials were inhibited by nifedipine. In the presence of L-NNA, sodium nitroprusside decreased the frequency of the action potentials without changing the resting membrane potential. These results indicated that contraction of rat portal vein circular muscles is inhibited tonically by nitric oxide, at least partly through inhibition of electrical activity.     

The role of alpha1-adrenoceptors in the clonidine-induced contraction and relaxation of rat aorta

Clonidine causes dilatation of the aorta in the presence of endothelium, while it causes contraction of the aorta in the absence of endothelium. The present study was carried out to clarify the role of alpha-1-adrenoceptors in the vascular action of clonidine. The aortic rings were suspended in Krebs-Henseleit (K-H) medium, and the effects of alpha-1- and alpha-2-adrenoceptor antagonists on the clonidine-induced contractions were measured. Moreover, the role of the phosphatidylinositol (PI) response was examined. The aortic slices were incubated in K-H medium containing, [3H]myo-inositol and clonidine. The formation of [3H]inositol monophosphate (IP1) was measured with a liquid scintillation counter. Clonidine caused contraction of the aorta in the absence of endothelium, in a dose-dependent manner. This contraction was inhibited by antagonists in the following order of the potency: prazosin > phentolamine > spiperone > urapidil = yohimbine > L-659066 > atipamezole. On the other hand, clonidine inhibited norepinephrine (NE)-induced contraction in the aorta in the absence and in the presence of endothelium. Clonidine enhanced IP1 accumulation in the aorta in the absence of endothelium, whereas it inhibited NE-induced IP1 accumulation in the aorta. The present results show that alpha-1-adrenoceptors are probably involved in the clonidine-induced contraction and relaxation of the rat aorta.     

Effects of neuropeptide Y (NPY) on mechanical activity and neurotransmission in the heart, vas deferens and urinary bladder of the guinea-pig

The effects of preincubation for 10 min with synthetic porcine neuropeptide Y (NPY) on muscle tone and autonomic transmission in the guinea-pig right atrium, vas deferens, urinary bladder, portal vein and trachea were analysed in vitro. NPY induced a metoprolol-resistant, long-lasting, positive inotropic and chronotropic effect per se in the spontaneously beating right atrium. Furthermore, NPY caused a reversible inhibition of both the metoprolol and atropine-sensitive auricle responses to field stimulation (2 Hz or 4 Hz for 2 s) without affecting the response to exogenous noradrenaline (NA) or acetylcholine (ACh). NPY did not induce any contraction of the vas deferens, but inhibited both the rapid twitch response and the sustained tonic contraction induced by field stimulation. The NPY-induced inhibition of the tonic contraction was more long-lasting than that of the twitch response. The tonic contraction was blocked by phentolamine and the twitch response by alpha-, beta-methylene ATP tachyphylaxis. NPY did not inhibit the contractile effects of NA, ATP or alpha-, beta-methylene ATP. NPY also induced a reversible reduction of the non-cholinergic, non-adrenergic contractile response to field stimulation of the urinary bladder. In the portal vein, NPY (up to 5 X 10(-7) M) did not inhibit the spontaneous motility or the phentolamine-sensitive contractile responses to field stimulation and NA. The atropine-sensitive contraction of the trachea or the non-adrenergic, non-cholinergic relaxation induced by field stimulation were not significantly influenced by NPY in doses up to 5 X 10(-7) M.(ABSTRACT TRUNCATED AT 250 WORDS)     

Electrical and mechanical properties of spontaneous contraction in hypertensive rat portal vein

Contractile and electrical activities of longitudinal smooth muscle of portal vein from normotensive Wistar Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) were compared. Amplitude and duration of spontaneous contraction of SHRSP portal vein were greater than those of WKY portal vein. No significant differences were observed in the resting membrane potentials between these preparations. Spontaneous spike activity appeared as a form of bursts. Duration of the burst and the number of spikes in each burst was greater in the portal vein of SHRSP than that of WKY. The amplitude of phasic and tonic components of K-contracture was also greater in SHRSP portal vein. Adrenergic and cholinergic nerves were not involved in the differences in contractions of the portal vein of these animal strains. Cross-sectional area of the longitudinal muscle layer was greater in SHRSP portal vein. These results suggest that the differences in spontaneous electrical activity are the cause of the differences in force and duration of the spontaneous contraction of portal vein from WKY and SHRSP, although the difference in excitation-contraction coupling of smooth muscle may be involved in much less extent.     

Mechanisms involved in contraction of smooth muscles of the rat portal vein as induced by sodium depletion

Responses to external Na depletion were investigated in the rat portal vein, using a microelectrode and an isometric force transducer. Mechanical response to Na depletion was characterized by a large tonic contraction with phasic contractions in a choline solution, and by phasic contractions without a tonic contraction in a Li solution. An identical depolarization of the membrane occurred in either choline or Li solution. After the tonic contraction was established in the Na-free choline-solution, the vein all but completely relaxed with partial re-admission of Na, while maintaining constant the concentration of choline. The Na-free choline-solution at 13 degrees C did not induce the tonic contraction. In nominal Ca-free solution, no contraction occurred with a depletion of Na. A tonic contraction was also induced by Na depletion in the presence of Mn but not Ca. It is concluded that with depletion of both external and internal Na, Ca and Mn may enter the cell through channels usually occupied by Na.     

Mechanisms underlying pre- and postjunctional effects of neuropeptide Y in sympathetic vascular control

The effects of porcine neuropeptide Y (NPY) regarding sympathetic vascular control were studied in vitro on isolated rat blood vessels. The 10(-9)M NPY enhanced (about two-fold) the contractile responses to transmural nerve stimulation (TNS), noradrenaline (NA) and adrenaline (about two-fold) in the femoral artery. Higher concentrations of NPY (greater than 10(-8)M) caused an adrenoceptor-resistant contraction per se. The TNS-evoked [3H]NA efflux was significantly reduced by NPY in a concentration-dependent manner (threshold 10(-9)M). The calcium antagonist, nifedipine, abolished the contractile effects of NPY and the NPY-induced enhancement of NA contractions but did not influence the prejunctional inhibition of [3H]NA release. Receptor-binding studies showed that the ratio of alpha 1-to alpha 2-adrenoceptors in the femoral artery was 30:1. The NPY did not cause any detectable change in the number of alpha 1-or alpha 2-adrenoceptor binding sites or in the affinity of alpha 2-binding sites, as revealed by prazosin- and clonidine-binding, respectively. The NPY also inhibited the TNS-evoked [3H]NA release (by 42-86%) in the superior mesenteric and basilar arteries and in femoral and portal veins. The NPY still depressed TNS-evoked [3H]NA secretion from the portal vein in the presence of phentolamine. The NPY caused a clear-cut contraction in the basilar artery, increased the contractile force of spontaneous contractions in the portal vein, while only weak responses were observed in the superior mesenteric artery and femoral vein. The NA-induced contraction was markedly enhanced by NPY in the superior mesenteric artery, only slightly enhanced in the portal vein and uninfluenced in the femoral vein. In conclusion, in all blood vessels tested, NPY depresses the TNS-evoked [3H]NA secretion via a nifedipine-resistant action. Furthermore, NPY exerts a variable, Ca2+-dependent vasoconstrictor effect and enhancement of NA and TNS contractions.     

Inhibitory actions of indomethacin on electrical and mechanical responses produced by nerve stimulation in circular smooth muscle of the guinea-pig gastric fundus.

The effects of indomethacin on electrical and mechanical responses produced by transmural nerve stimulation (TNS) were investigated in isolated circular smooth muscle of the guinea-pig gastric fundus. TNS evoked a cholinergic excitatory junction potential (e.j.p.). The e.j.p.s were inhibited by 1-10 microM indomethacin, in a concentration-dependent manner, with no marked alteration of the resting membrane potential. Exogenously applied acetylcholine caused a depolarization of the membrane that was not altered by indomethacin. TNS evoked a cholinergic twitch contraction at low frequencies (0.1 Hz). A train of TNS's at high frequency (1 Hz) produced a transient contraction with a subsequent sustained relaxation. Indomethacin reduced the resting tension and inhibited these TNS-induced contractions. Application of Nomega-nitro-L-arginine (NOLA), an inhibitor of nitric oxide (NO) synthesis, increased the amplitude of twitch contractions, and altered transient contractions to tetanic contractions during TNS at a frequency of 1 Hz, also with an increased amplitude. In the presence of NOLA, indomethacin (5 microM) again reduced the resting tension and inhibited TNS-induced contractions. This inhibition was greater for twitch contractions than for tetanic contractions. Nifedipine reduced the TNS-induced contractions, while addition of indomethacin further reduced the amplitude of contractions. Contractions produced by low concentrations of acetylcholine (0.1 microM) were inhibited by indomethacin, while those produced by 1 microM were not. These results indicate that the inhibitory actions of indomethacin on TNS-induced contractions do not involve enhanced production of NO or selective inhibition of voltage-gated Ca-channels. Prejunctional autoregulatory mechanisms may also not be altered by indomethacin. As indomethacin inhibits the enzyme cyclooxygenase, it is speculated that endogenously produced prostaglandins exert excitatory actions on gastric smooth muscle, and act mainly postjunctionally to facilitate spontaneous and neurogenic electrical and mechanical activity.     

Fine structure of the mouse portal vein in relation to its peristaltic movement

The hepatic portal vein has been known to make a spontaneous peristaltic movement in some mammals, including the mouse and rat. To investigate the fine structure of the portal vein in relation to its physiological characteristics, we observed the mouse portal vein by using various histological techniques including conventional light microscopy, videomicroscopy, transmission and scanning electron microscopy, and real-time confocal laser scanning microscopy. The mouse hepatic portal vein was provided with a spiral fold which was produced by the inner layer, i.e. the endothelium and smooth muscles of the wall protruding into the lumen. Longitudinal smooth muscle cells spanned the interval of the fold, like a spirally arranged palisade around the vessel wall. The longitudinal muscle fibers ended at the spiral fold, being partly connected with a network of irregularly shaped smooth muscle cells. This network, hitherto unknown, was recognized to be restricted to the fold in distribution and characterized by numerous gap junctions connecting the muscle cells. Real-time confocal laser scanning microscopy using a Ca2+ sensitive fluorescent dye revealed that a transient and periodic increase in Ca2+ concentration occurred in the longitudinal smooth muscle cells and was transmitted spirally from the intestinal to the hepatic side. These findings indicate that, during the peristaltic movement, the contraction of smooth muscle cells is transmitted along the longitudinal smooth muscles of the portal vein wall toward the liver, presumably controlled by the network of the irregularly-shaped smooth muscle cells in the fold of the portal vein. Light microscopic observation in some specimens indicated an occurrence of cardiac muscle cells outside the smooth muscle layer. Restricted to the site of the porta hepatis in distribution, their involvement in the peristaltic contraction of the portal vein seemed unlikely.     

The mechanical response of rat myometrium to adenosine triphosphate in Ca-free solution

The contractile response of the myometrial longitudinal muscle of pregnant and estrogen-treated rats to adenosine triphosphate (ATP) was investigated. ATP (10(-5)-4 X 10(-3) M) added to Krebs solution caused a generation of spontaneous activity consisting of phasic contractions and an elevation of muscle tone in a dose-dependent manner. Effects of the "test solution" consisting of isotonic K, 4(mM) ATP, 4 Mg, 1-20 EGTA, 20 tris(hydroxymethyl)aminomethan (Tris) maleate (pH 6.8) on the contractile response was then investigated. An initial phasic and a following tonic contraction was evoked by the application of the test solution, when applied after the equilibration of the muscle with Krebs solution. ATP was proved to be an agonist to evoke the Ca-free contraction. The phasic contraction was depressed when the muscle was incubated with Ca-free Krebs solution. The amplitude of the tonic contraction became progressively larger when application of the test solution was repeated. The amplitude was 15-70% as large as the tonic component of the K-contracture induced by 40 mM K. Theophylline (10 mM), 0.1 mM papaverine and 1 microM isoprenaline nearly abolished, and 1 mM cAMP partly depressed the tonic contraction of K-contracture, whereas the tonic contraction induced by the test solution was unaffected. A calmodulin antagonist, W-7 (100 microM) strongly suppressed both the K-contracture and the contraction induced by the test solution, whereas trifluoperazine (10-200 microM) preferentially depressed the K-contracture. The tonic contraction induced by the test solution was strongly depressed when Mg was removed and 20 mM EDTA was applied. From these results, it is discussed that some Ca-independent process is involved in the generation of tonic component of Ca-free contraction developed by the application of the test solution.     

Evidence that ATP released from the postsynaptic site by noradrenaline, is involved in mechanical responses of guinea-pig vas deferens: cascade transmission.

The release of endogenous ATP and [3H]noradrenaline, and the mechanical response of the guinea-pig vas deferens to field stimulation of its motor nerves were examined using a perfusion system. The release of ATP at rest was 0.83 +/- 0.13 pmol/g per min, and ATP released by field stimulation (8 Hz, 480 shocks) was 5.47 +/- 1.23 pmol/g. The evoked release was completely inhibited when Ca2+ was removed and 1 mM EGTA was added, or by 1 microM tetrodotoxin. The release of ATP and [3H]noradrenaline in response to field stimulation was constant with an S2/S1 ratio of 1.10 +/- 0.11 for ATP and 0.92 +/- 0.03 for [3H]noradrenaline, respectively (where S1 and S2 are stimulation periods). Prazosin (1 microM), a potent alpha 1-adrenoceptor antagonist, significantly reduced the stimulation-evoked release of ATP by 75% and significantly reduced both mechanical twitch and tonic responses, but enhanced the release of [3H]noradrenaline. This finding indicates that there is an alpha 1-adrenoceptor-mediated release of endogenous ATP. However, the prazosin-insensitive portion of ATP release (25%) is considered to be of presynaptic origin. The stimulation of alpha 1-adrenoceptors by 1-noradrenaline or methoxamine in concentrations ranging from 10 to 100 microM resulted in a concentration-dependent release of ATP and a biphasic contraction of the vas deferens: a twitch response was followed by a tonic contraction. Prazosin (1 microM) completely prevented the effect of 1-noradrenaline or methoxamine on both ATP release and mechanical response. When Ca2+ was omitted and EGTA (1 mM) was added, 1-noradrenaline was still able to release ATP but failed to produce contraction. Nifedipine, a Ca-channel and ATP receptor antagonist, reduced the twitch contraction and enhanced the release of ATP from muscle in response to noradrenaline administration. This finding indicates that the release of ATP from the muscle is not linked to mechanical contraction. When the vas deferens was made deficient in noradrenaline by 6-hydroxydopamine pretreatment (100 + 250 mg/kg, i.p.), electrical field stimulation failed to release [3H]noradrenaline and ATP. Under these conditions, exogenous 1-noradrenaline was much more effective in releasing ATP from the smooth muscle, and producing twitch responses, followed by a tonic contraction. After reserpine pretreatment (2 x 5 mg/kg, i.p.), the field stimulation-evoked release of ATP and both phases of contraction were markedly reduced.(ABSTRACT TRUNCATED AT 400 WORDS)     

Concentrations and contractile effects of substance P in the human ampullary-isthmic junction

From 20 women undergoing hysterectomy, strip preparations were isolated from the outer, longitudinal and the inner, circular smooth muscle layer of the ampullary-isthmic junction (AIJ), together with small arterial segments dissected as ring preparations from the root of the mesosalpinx. The specimens were mounted in organ baths and isometric tension was recorded. In addition, tissue concentrations of substance P (SP) in the ampulla, AIJ and utero-tubal junction were determined by radioimmunoassay. Tissue concentrations of SP expressed as pmol X g tissue-1 (wet weight, +/- SE) amounted to 3.09 +/- 1.40 in the utero-tubal junction, 1.08 +/- 0.299 in the AIJ and 0.742 +/- 0.299 in the ampulla. In strips of circular muscle, SP at concentrations of 10(-7) -3 X 10(-6) mol X l-1 elicited a combined phasic and tonic response and in longitudinal muscle a mainly tonic contraction was produced. In both tissues, contractions elicited by SP were rapidly abolished in calcium-free medium. Nifedipine abolished the phasic contraction elicited in circular muscle by SP while the tonic response was resistant. The contraction in longitudinal muscle was reduced by 20-30%. Vasoactive intestinal polypeptide (VIP) decreased tension in preparations contracted by SP, prostaglandin F2 alpha and K+-depolarization (124 mmol X l(-1). In unstimulated oviductal arterial preparations, SP had no effect, while the peptide induced a transient relaxation of noradrenaline contracted preparations, and slightly decreased tension of K+-depolarized vessels. The results suggest that SP may be involved in the control of motility of the human AIJ.     

Mechanical and electrical responses modulated by excitation of inhibitory nerves during stimulation with high-potassium solutions in circular smooth muscle of the rabbit rectum.

The properties of the mechanical responses produced by solutions containing high concentrations of potassium ion (high-K solution, [K(+)](o) = 9-27 mM) were investigated in circular smooth muscle preparations isolated from the rabbit rectum. Isometric recording of mechanical responses of the muscle revealed spontaneous contractions, which successively decreased and finally disappeared in most preparations. Stimulation of the smooth muscle with high-K solutions elicited an increase in both amplitude and frequency of twitch contractions (sustained component), with about a 2 min delay in the beginning (initial inhibition), and a transient large contraction shortly after the cessation of stimulation (after contraction). Transmural nerve stimulation (TNS) with electrical pulses for 1 min at 1 Hz frequency produced a sustained inhibition, but a transient contraction followed after termination of TNS. In the presence of tetrodotoxin (TTX), the TNS-induced responses were abolished, while a high-K solution elicited increased twitch contractions with a short delay and abolished the after contraction. Suramin produced effects similar to TTX on the responses produced by high-K solutions or TNS, but this was not the case for atropine, guanethidine or N(omega)-nitro-L-arginine (L-NA). Recording membrane potentials with microelectrodes revealed that TNS evoked an inhibitory junction potential (i.j.p.) which was non-adrenergic, non-cholinergic and non-nitrergic in nature. High-K solutions elicited a tri-phasic change in the membrane potential; an initial hyperpolarization, followed by a sustained depolarization and finally a transient depolarization on cessation of high-K stimulation. TTX or suramin inhibited the i.j.p.s and altered the tri-phasic change in the membrane potential produced by a high-K solution to a mono-phasic depolarization. No significant modulation of electrical responses of the membrane induced by TNS or high-K solution was elicited by atropine, guanethidine or L-NA. The results indicated that the circular smooth muscle of the rabbit rectum is innervated by inhibitory nerves, and that stimulation with high-K solutions caused inhibitory neuronal modulation of both electrical and mechanical responses of the smooth muscle, in a suramin-sensitive way.     

Influences of endothelium on the time course of noradrenaline-, 5-HT-, prostaglandin F2alpha- and high-K+-induced contractions in aortae of WKY and SHRSP.

Influences of endothelium on contraction of aortic smooth muscle by various agents were studied and those in the preparations from Wistar Kyoto rat (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) were compared. Endothelium depressed the contractions induced by noradrenaline and 5-hydroxytryptamine (5-HT). The time course of the contraction was bi- or tri-phasic in the former and slow rising monophasic in the latter. On the other hand, the depression was weaker in the contraction by prostaglandin F2alpha- and high K+. The depression was blocked by the removal of endothelium or in the presence of Nomega-nitro-L-arginine (L-NNA), indicating that nitric oxide (NO) released from endothelium was responsible for the inhibition. The inhibition was weaker in the preparation from SHRSP when compared to that in the preparation from WKY. Relaxation by acetylcholine (ACh) of the preparation precontracted in the presence of respective contractile agent was impaired in the preparation from SHRSP. It is concluded that mode of inhibition of the contraction varies depending on the agents used to initiate the contraction, i.e. depending on the mode of the release of NO. In the preparation from SHRSP, the influence of endothelium is impaired due to the reduced release of NO.     

Direct and indirect contractile responses of the human vas deferens and actions of noradrenaline and of calcium antagonists

Contractile responses of the isolated human vas deferens, obtained from vasectomy operations, were measured. Large single electrical shocks gave a twitch response with short latency (0.36 s) which was insensitive to prazosin (5 microM) or TTX (0.2 microM) and was thus identified as due to direct muscle stimulation. A train of 100 low intensity shocks gave a response with a longer latency (1.9 s) which was substantially sensitive to both prazosin and TTX; we assume this response is dominated by an indirect nerve-induced contraction. Relaxations, presumably caused by activation of circular muscle, were recorded from regions of some preparations both by direct and indirect stimulation. Noradrenaline (10-20 microM) induced a tonic contracture, spontaneous contractions and a large potentiation of the response to direct stimulation--but not to indirect stimulation implying a strong presynaptic inhibition. Noradrenaline also speeded the relaxation from contractions. Verapamil (1-100 microM) and nifedipine had no effect on the direct responses but verapamil (10 microM) inhibited the indirect response. Calcium removal prevented most, and 5 mM-EDTA all, of the direct response. However, even with EDTA, noradrenaline was able to support spontaneous and stimulus-induced contractions. Thus contraction of the vas, though sustained by external calcium, does not appear to directly depend on it.     

Phasic and tonic contractions of rabbit intestinal muscle.

The effects of a single electrical stimulus upon the longitudinal and circular muscles of rabbit small intestine were investigated and the differentiation between phasic and tonic contractions was examined. A weak stimulus caused a phasic contraction and a stronger stimulus evoked both phasic and tonic contractions. The evoked phasic contraction was of an all-or-nothing nature, while the evoked tonic contraction was graded. Strength-duration curves of longitudinal and circular muscles were obts of rheobase and chronaxie. In the presence of verapamil or methoxyverapamil (D 600) the tonic contraction, but not the phasic contraction, could be evoked by a single strong stimulus, while in the presence of sodium nitroprusside the phasic contraction, but not the tonic contraction, could be provoked by a single strong stimulus. The phasic and tonic contractions differed in their strength-duration curves and drug responses.     

Regulatory mechanisms for the spontaneous contractile force and frequency of the rat portal vein]

By the extracellular recording technique, the action potentials and spontaneous contractions of the isolated rat longitudinal portal vein strips were simultaneously recorded in the presence of varying concentrations of electrolytes, various vasoactive agents, and hormones, and the mechanisms regulating the force and frequency of spontaneous contraction of the vascular smooth muscle were investigated. A longitudinal stretch (-200% of the initial length), a higher [K+]0 (-30 mM), a lower [Na+]0, epinephrine, acetylcholine or ouabain increased both force and frequency of the phasic contractions. A lower [Na+]0 or ouabain raised basal tone of the muscle in addition to the above effects. A higher [Ca++]0 increased the contractile force, while it decreased the frequency. A lower [Ca++]0, calcium channel blocker or VIP reduced the contractile force but increased the contractile frequency. A bigger force and a higher frequency of the phasic contractions were exhibited in the portal vein strips isolated from rats with CCl4-induced experimental portal vein hypertension. They were similar to responses of the strips from the intact rats to stretch, ouabain, or higher [Ca++]0 in the presence of lower [Na+]0. These results suggest that an initiation and force of the phasic and tonic contractions depend on extracellular Ca++ concentration and influx of Ca++, and frequency of the phasic contractions, mainly on membrane potential rather than on extracellular Ca++ concentration. In the portal hypertension, permeability of the cell membrane to Ca++ is possibly increased.     

Influence of extracellular calcium and nifedipine on alpha 1- and alpha 2-adrenoceptor-mediated contractile responses in isolated rat and cat cerebral and mesenteric arteries

The influence of extracellular Ca2+ and nifedipine on contractile responses to 10 microM noradrenaline (NA) was investigated in isolated rat and cat middle cerebral (RCA, CCA) and mesenteric (RMA, CMA) arteries. In the CCA (containing predominantly alpha 2-adrenoceptors), the NA-induced contractions developed considerably more slowly than in the RCA, RMA (containing mainly alpha 1-adrenoceptors) and CMA (sensitive to both alpha 1- and alpha 2-adrenoceptor selective antagonists). The tonic component of the NA-induced contraction in the four types of artery was substantially suppressed after only short periods in Ca2+-free solution. In each type of artery, excluding the CCA, the contractile response to 124 mM K+ was more sensitive to Ca2+ deprivation than that to NA. This suggests that NA, besides mobilizing extracellular Ca2+, can also release Ca2+ from an intracellular pool in the RCA, RMA and CMA, but not in the CCA. Thus, alpha 1-adrenoceptor-mediated contractions in the RCA and RMA seem to depend on both Ca2+ influx and intracellular Ca2+ release, whereas alpha 2-adrenoceptor-mediated contractile responses in the CCA appear to rely almost entirely on Ca2+ influx. Both the maximum response and the tonic component of the NA-induced contraction were significantly more sensitive to nifedipine in the CCA than in the RCA. In comparison with the NA-induced contractions in these arteries, those in the RMA and CMA were relatively resistant to nifedipine. In the CCA exposed to NA in Ca2+-free medium, nifedipine almost abolished the contraction induced by re-addition of Ca2+, whereas in the other types of artery, Ca2+ re-application evoked a significant contraction also in the presence of the drug. The differential effects of nifedipine presumably reflect differences between the arteries, not only in the relative contribution of Ca2+ influx and intracellular Ca2+ release to the contractile activation, but also in the nifedipine sensitivity of the Ca2+ entry pathways utilized by NA. It is concluded that the mechanisms through which NA induces contraction seem to be related both to the subtype of alpha-adrenoceptor stimulated by NA and to the type of vessel studied.     

Influence of 2,3-diphosphoglycerate on the buffering properties of human blood

Summary The recruitment properties of single motor units from the human extensor indicis muscle were investigated during voluntary isometric contractions of different rate of rise but equal amplitude. Both the electrical and the contractile events associated with recruitment were analyzed. The threshold force of recruitment (measured as the total muscle force at firing onset) decreased with increasing rate of rise of isometric tension. This was consistently found for all units. Differences between low and high threshold units indicating a preferential tonic or phasic mode of activation were not observed.The contractile events associated with recruitment were analyzed analoguously to the electrical events. For this purpose, muscle force was measured at the time of the first twitch as it was measured at the time of the first spike. This separate measurement of the electrical and mechanical recruitment of a unit is necessary, because during a change of muscle force, force is different at firing onset and during the subsequent twitch contraction. Muscle tension at the time of the peak of the first twitch contraction was calculated from measurements of the twitch contraction time of the single motor units.In contrast to firing onset, the peak of the first twitch of a motor unit occurs at approximately the same muscle tension no matter how fast the contraction is performed.This is the consequence of the result that the average decrease of the threshold force of recruitment at successively faster contraction has the same value as the corresponding increase of total muscle force during the mean contraction time of the motor units. On the basis of this precise matching between these two changes, the mechanical recruitment of motor units occurs at approximately the same force level irrespective of the rate of rise of tension.Supported by Deutsche Forschungsgemeinschaft, SFB 70.The experiments reported in this paper are adherent with the principles embodied in the declarations of helsinki. Informed consense was obtained from each subject and they were aware that they could withdraw from the experiment at any time. Pain was minimal equivalent to that experienced during routine clinical electromyography.     

Relaxation of canine saphenous vein following brief transmural nerve stimulation

The ability of electrical stimulation to cause relaxation in the canine saphenous vein was evaluated. Rings of vein were studied isometrically in organ chambers containing physiological salt solution. Prostaglandin F2 alpha produced stable contractions, during which brief periods of electrical stimulation caused further contraction. With cessation of the electrical stimulation, tension transiently decreased to a level below that observed prior to the stimulation (undershoot). This poststimulation undershoot was blocked by tetrodotoxin, phentolamine, ouabain, and potassium-free solution; it was not affected by atropine, cimetidine, indomethacin, ketanserin, methysergide, propranolol, pyrilamine, or removal of the endothelium. Undershoot did not occur following electrical stimulation during contractions evoked by norepinephrine. During superfusion with PGF2 alpha, a brief exposure to exogenous norepinephrine caused a transient contraction followed by a subsequent undershoot. These results suggest that 1) the interaction of norepinephrine with postjunctional alpha-adrenoceptors on vascular smooth muscle leads to an increase in the activity of Na+-K+-ATPase, and 2) increased activity of Na+-K+-ATPase is responsible for the poststimulation undershoot.