Transmitter release modulated by isoprenaline in the dog isolated mesenteric vein

1. The effects of isoprenaline on the release of transmitter substances from perivascular adrenergic nerves were estimated from the excitatory junction potential (ejp) and outflow of noradrenaline in the dog mesenteric vein. 2. Isoprenaline increased the ejp amplitude and decreased the evoked release of noradrenaline. Yohimbine potentiated the former and converted the latter to an increased outflow. 3. Therefore, stimulation of prejunctional beta-adrenoceptor by isoprenaline increases the release of noradrenaline from perivascular adrenergic nerves. 4. Possible involvement of prejunctional alpha-adrenoceptors in the isoprenaline-induced modulation of transmitter release was also suggested.     

Facilitation of nerve stimulation evoked noradrenaline overflow by isoprenaline but not by circulating adrenaline in the dog in vivo

The influence of isoprenaline and adrenaline on the overflow of endogenous noradrenaline evoked by sympathetic nerve stimulation was studied in canine blood perfused gracilis muscle in situ. Neuronal uptake was inhibited by desipramine. Local i.a. infusions of isoprenaline enhanced stimulation evoked noradrenaline overflow by 32 +/- 10% (P less than 0.05), indicating the existence of prejunctional facilitatory beta-adrenoceptors. This effect of isoprenaline was not antagonized by beta 1-adrenoceptor blockade and does not seem to be related to the vasodilatation caused by isoprenaline. In a second series of experiments circulating adrenaline levels were raised by i.v. infusions from basal levels of 0.4 +/- 0.2 nM to 1.7 +/- 0.2 and 6.3 +/- 0.6 nM, respectively, in arterial plasma. Adrenaline elicited vasodilatation in the gracilis muscle (19 +/- 3 and 28 +/- 5% increases in vascular conductance, respectively), indicating activation of postjunctional beta 2-adrenoceptors, without influencing nerve stimulation evoked noradrenaline overflow. Thus, our results support the existence of a prejunctional beta 2-adrenoceptor mediated mechanism facilitating noradrenaline release in vivo, but provide no evidence to support the idea that physiologically relevant increases in circulating adrenaline levels enhance noradrenergic neurotransmission in skeletal muscle.     

Changes in the electrical activity of the rabbit proximal colon in vivo by stimulation of the vagus and splanchnic nerves]

1. Using extracellular electrodes placed on the serosa, we recorded the modifications of the electrical activity of the colonic muslce fibers caused by the stimulation of vagal and splanchnic nerve fibers. 2. Vagal stimulation produces two types of junction potentials: excitatory junction potentials (EJPs) and inhibitory junction potentials (IJPs). The IJPs are elicited by stimulation of vagal fibers which innervate intramural non-adrenergic inhibitory neurons. 3. The conduction velocity of the nerve impulse along the vagal pre-ganglionic fibers is 1.01 m/sec for excitatory fibers and 0.5. m/sec for inhibitory fibers. 4. Splanchnic fiber stimulation causes EJP disappearance, blocking transmission between preganglionic fibers and intramural excitatory neurons, and a decrease in IJP amplitude that most likely indicates a previous hyperpolarization of the smooth muscle. 5. IJP persistence during splanchnic stimulation proves that sympathetic inhibition does not modify the transmission of the vagal influx onto the non-adrenergic inhibitory neurons of the intramural plexuses. 6. Through a comparative study of proximal and distal colonic innervation, we are able to show that there is a similar organization of both regions, that is a double inhibitory innervation: an adrenergic one of a sympathetic origin, and a non adrenergic one of a parasympathetic origin.     

The effect of decentralisation or chronic hypogastric nerve stimulation in vivo on the innervation and responses of the guinea-pig vas deferens

Summary The physiological, pharmacological and morphological characteristics of guinea-pig vas deferens supplied by hypogastric nerves rendered inactive by decentralisation were compared with those of vas deferens in which the nerve supply had been chronically stimulated for 3–9 days using implanted electrodes. No change was seen in decentralised preparations prior to 7 days, but from 8–15 days, increased sensitivity to application of noradrenaline in vitro was observed, which was shown to be related to reduced transmitter uptake by nerve terminals as well as to an increase in postjunctional sensitivity; there was also increased fatigability 7–14 days following decentralisation. Continuous stimulation of hypogastric nerves at 2 Hz for 4–8 h daily for 4–8 days resulted in enhanced transmitter uptake and reduced responses to noradrenaline; this was associated with a slight increase in noradrenaline content and a faster adrenergic neuromuscular response with a shorter latency. No appreciable changes in nerve or muscle structure studied by electron microscopy were observed following decentralisation, but there was an increase of between 12.5 and 29.6% in the number of close (< 100 nm) neuromuscular junctions following chronic stimulation for 8 days.     

Activities of prostaglandins and prostaglandin endoperoxides at adrenergic neuroeffector junctions.

The results presented in this paper indicate that: 1. The prostaglandin synthesis inhibitor, indomethacin, increases noradrenaline turnover in a variety of rat organs. This observation increases the probability that prostaglandins are involved in the control of adrenergic neurotransmission in vivo. 2. Administration of endoperoxides inhibits the release of noradrenaline from adrenergic nerve terminals. The effect can be explained, however, at least in part, by formation of degradation products, presumably mainly prostaglandin E2. 3. Prostaglandin F2 alpha enhances smooth muscle responses to adrenergic nerve stimulation in rabbit heart and guinea pig vas deferens. These actions must be considered prostjunctional, since the release of noradrenaline is unchanged or depressed.     

The effect of cocaine, reserpine, and 6-hydroxydopamine on the response of the perfused central artery of the rabbit ear to sympathomimetic amines.

The perfused central artery of the rabbit ear was less sensitive to extraluminal than to intraluminal noradrenaline, but the reverse was true for metaraminol, methoxamine, metanephrine, and isoproterenol. No difference was noted between the extraluminal and intraluminal potency of phenylephrine. Cocaine potentiated the effect of extraluminal and intraluminal noradrenaline, but decreased that of intraluminal phenylephrine. Irrespective of the route of administration, the constrictor potencies of the sympathomimetic amines were not affected by cocaine. Arteries of reserpine-treated rabbits were supersensitive to extraluminally and intraluminally applied noradrenaline and phenylephrine, but they were not supersensitive to metaraminol. 6-Hydroxydopamine effectively destroyed adrenergic nerve endings of the central ear artery and increased its responses to both extraluminal and intraluminal noradrenaline and phenylephrine. However, only the constrictor potencies of intraluminally applied metaraminol and methoxamine were enhanced by 6-hydroxydopamine. The apparent discrepancies between the results obtained by various procedures that eliminate or impair the nerve uptake process suggest that the difference in the constrictor potency of extraluminal and intraluminal sympathomimetic amines is probably unrelated to their uptake by nerves located in the adventitio-medial junction of the artery.     

Contractile responses to adrenergic nerve stimulation are enhanced with removal of endothelium in rat caudal artery

Removal of the endothelium from isolated perfused rat caudal arteries produced a two fold increase in the contractile response to transmural nerve stimulation. Pretreatment with 6-hydroxydopamine eliminated the contractile response to adrenergic nerve stimulation but failed to uncover any vasodilatory effect of electrical stimulation, either directly on smooth muscle or via non-adrenergic nerves. Endothelial removal also produced two and four fold enhancement of the contractile responses to the selective alpha 1- and alpha 2-adrenoceptor agonists methoxamine and B-HT 920. However, pKB values for prazosin and yohimbine versus both agonists indicate that both methoxamine and B-HT 920 are acting primarily at alpha 1-adrenoceptors in this tissue. These results provide evidence that endothelial factors released either at basal levels or by the stimulation of agonists play a significant physiological role in modifying the contractile responses of blood vessels.     

Effect of cocaine on responses of mouse phrenic nerve-diaphragm preparation

Effects of 5 to 40 microM cocaine on the compound action potential (AP) and tension responses of the mouse phrenic nerve-diaphragm preparation were monitored following nerve and muscle stimulation at 37 degrees C. Cocaine caused concentration dependent reduction in amplitude of the nerve AP, muscle AP, and tension response to a single nerve stimulus, and greater reduction in amplitude of these responses to repetitive nerve stimuli at 100 Hz for 0.5 sec. Cocaine caused similar reduction in the muscle AP and tension responses to direct muscle stimulation in the presence or absence of curare, and markedly reduced the overshoot, total potential, and maximum rate of rise and fall of intracellularly recorded muscle AP, without affecting the resting potential, or the contracture responses evoked by caffeine. These results indicate that cocaine reduces skeletal muscle function by reducing the excitability of muscle and nerve membranes, without significantly affecting neuromuscular transmission, excitation-contraction coupling or contractility.     

Neurogenic constrictor response of isolated small renal arteries in rats after 2-week simulated microgravity

The study was aimed at investigation of the effects of 2-week tail suspension upon the constrictor responses of isolated small renal arteries in rats. 1st-2nd-order branches of renal artery were perfused with saline under the constant flow conditions. Constrictor responses to electrical stimulation of periarterial nerves, noradrenaline and serotonin were investigated. In post-suspension rats as compared to controls the response to nerve stimulation was slightly reduced during 15-Hz stimulation, but similar at smaller frequencies. Thus, simulated microgravity has no prominent effect of neurogenic responses of renal vessels, in agreement with non-changed density of periarterial adrenergic nerve plexus. Along with that, in post-suspension rats impairment of prejunctional sympathetic mechanisms might be compensated by augmented sensitivity of vascular smooth muscle to vasoconstrictors.     

The effect of caesium on adrenergic transmission in rabbit vas deferens.

The effect of caesium on the responses of rabbit vas deferens to transmural stimulation was investigated. The tissue responded to transmural stimulation with a phasic spike contraction followed bya sustained contractile response. The sustained response was inhibited by phentolamine and guanethidine and thus apparently results from noradrenaline release from adrenergic nerves. Addition of 2-5mM Cs+ greatly potentiated this secondary response without altering the sensitivity of the tissue to added (minus)-noradrenaline. This potentiation was not due to Cs+ decreasing the neuronal uptake of noradrenaline, or by Cs+ altering prostaglandin synthesis. Addition of 2mM Cs+ significantly increased the amount of (plus or minus)-[3-H] metaraminol released from tissues in response to transmural stimulation (5 Hz). It is suggested that caesium potentiated responses of rabbit vas deferens to transmural stimulation by increasing the amount of transmitter released per nerve impulse, possibly as a result of prolongation of the action potential.     

Occurrence of uranaffin-positive synaptic vesicles in both adrenergic and non-adrenergic nerves of the rat anococcygeus muscle

Summary The uranaffin reaction in rat anococcygeus muscle, which receives a dual innervation of both adrenergic and non-cholinergic, non-adrenergic nerves was examined. Dense reaction product was observed in the vesicular membranes and/or the cores of some synaptic vesicles in the adrenergic nerve terminals. Occasional vesicles were filled up with dense reaction product. In the prominent population of small clear vesicles, however, no dense reaction product was observed. The number of small granular vesicles in the adrenergic nerve terminals was markedly increased after the administration of 5-hydroxydopamine (5-OHDA). These granular vesicles were moderately stained with uranaffin deposit on the cores but their limiting membranes possessed no uranaffin deposit at all.In the non-adrenergic nerve terminals, on the other hand, uranaffin deposit of variable density was observed on the cores of large granular vesicles but never on their limiting membranes or on the small clear vesicles. There was no change in the axon profiles after the administration of 5-OHDA.The possible occurrence of purines in the cores of large granular vesicles in the non-adrenergic nerves is discussed.     

Distribution of myelinated nerves in ascending nerves and myenteric plexus of cat colon

The parts of the colon differ in motor function and in responses to extrinsic and intrinsic nerve stimulation. The distribution of myelinated nerve fibers in the colonic myenteric plexus is not known. Because these fibers might be largely extrinsic in origin, their distribution might indicate the domain of influence of extrinsic nerves and help to explain the different behaviors of the different parts of the colon. Myelinated fibers were examined by electron microscopy in cross sections of the ascending nerves and in myelin-stained whole-mount preparations in the colon. The ascending nerves are much like one another. They have the structure of peripheral nerves, not that of myenteric plexus. The proportion of myelinated fibers in the ascending nerves declines rostrad with no uniform change in total nerve fiber number. Cross-sectional areas of ascending nerves, 3,304 to 7,448 microns 2; total number of nerve fibers per profile, 703-2,651; and mean myelin coat thickness, 0.45 +/- 0.01 micron, do not change uniformly along the ascending nerves. Myelinated fibers are about 2% of total fibers in the extramural colonic nerves, 7-9% in the ascending nerves in the sigmoid colon, and 2-3% at the rostrad ends of the ascending nerves in the transverse colon. Blood vessels lie at the core of each ascending nerve and on the nerve sheath. Myelinated fibers in the ascending nerves degenerate after section of colonic branches of the pelvic plexus and after section of the pudendal nerves, indicating that myelinated nerves reach the colon through both pathways.(ABSTRACT TRUNCATED AT 250 WORDS)     

Two types of plasticity in the tentacle withdrawal reflex of Helix aspersa are dissociated by tissue location and response measure

The tentacle withdrawal reflex of the terrestrial snail Helix aspersa was studied in vitro. The reflex is evoked by mechanical stimulation of the nose. Lesion experiments showed that 45% to 75% of the response amplitude is attributable to peripheral pathways alone. The central contribution increases with increasing stimulus intensity.Repeated stimulation produced pure habituation at low stimulus strengths, and habituation mixed with intrinsic sensitization (warm-up effect) at high stimulus strengths. The simultaneous occurrence of habituation and sensitization is consistent with the dual process theory of plasticity. Additional results differentiate the two processes. Habituation can occur without the CNS, whereas intrinsic sensitization requires the CNS. Also, the two processes are differentially effective in their influences on response amplitude and duration: habituation is more effective in determining response amplitude, while sensitization is more effective in determining response duration.Although the establishment of sensitization requires the CNS, 81% of the memory for intrinsic sensitization was localized to the periphery, by lesion experiments. Extrinsic sensitization, caused by stimulation of the medial lip nerve, had similar behavioural effects and a similar memory locus. Both types of sensitization appear to be caused by neuromuscular facilitation mediated by a central pathway.Abbreviations CNS central nervous system - PNS peripheral nervous system - S-R stimulus-response - TRM tentacle retractor muscle     

NEUROCHEMICAL PROPERTIES OF ADRENERGIC NERVES REGENERATED AFTER 6-HYDROXYDOPAMINE

Abstract— The uptake-storage properties and synthesis of noradrenaline, and fluorescence morphology of adrenergic nerves which have been allowed to regenerate for 4 weeks after a chemical sympathectomy produced by 6-hydroxydopamine have been investigated in mouse iris and atrium. The regenerated nerve terminals displayed a lower formaldehyde-induced fluorescence intensity whereas the non-terminal axons exhibited a stronger fluorescence intensity and a more beaded appearance compared with mature nerves. The endogenous noradrenaline concentration after 6-hydroxydopamine was 30% in iris and 45% in atrium compared to control values. Recovery of [³H]noradrenaline uptake was found to be more rapid than that of endogenous noradrenaline concentration after the 6-hydroxydopamine treatment. [³H]Noradrenaline uptake in regenerating and adult mature nerves both obeyed Michaelis-Menten kinetics having identical K_m values. There was a close correlation between [³H]noradrenaline uptake and nerve density of adrenergic nerves regenerated after 6-hydroxydopamine. These results show that [³H]noradrenaline uptake is a better index for the number of regenerated nerve terminals than is the endogenous noradrenaline concentration. The retention of [³H]noradrenaline taken up and accumulated in vitro was about the same in regenerated and mature nerves, although a slight tendency to less effective retention was observed in the regenerated nerves. Subcellular distribution studies showed that relatively less [³H]noradrenaline was recovered in the microsomal fraction after 6-hydroxydopamine treatment. The formation of ¹⁴C-labelled catecholamines from [¹⁴C]DOPA was higher in regenerating nerves than indicated by the endogenous noradrenaline concentration but lower than that indicated by the [³H]noradrenaline. It is concluded that the regenerating nerves contain less endogenous noradrenaline than adult mature nerves and that the uptake mechanism develops promptly, whereas the development of the storage mechanism lags behind.     

Adrenergic component in the mechanism of action of sodium oxybutyrate]

The influence of sodium oxybutyrate on the adrenergic neurotransmitter content in the nerve fibers and the synaptic vesicles and the uptake of exogenous noradrenaline (NA) by the sympathetic nerves of rat Vas deferens was studied by spectrofluorimetry, fluorescent-histochemistry and cytochemical electron microscopy. Sodium oxybutirate failed to influence the stores of the adrenergic mediator, but was capable of blocking the uptake and accumulation of the exogenous NA.     

Action of a beta-bungarotoxin on autonomic ganglia and adrenergic neurotransmission.

A beta-bungarotoxin was isolated from the venom of Bungarus multicinctus by column chromatography on Sephadex G-50 and SP-Sephadex. The toxin produced presynaptic effects on neuromuscular transmission with characteristics similar to those described by others. In a sympathetic ganglion, the toxin increased spontaneous acetylcholine (ACh) release and decreased ACh release evoked by preganglionic nerve stimulation. The toxin did not block the response of isolated ileum to cholinergic nerve stimulation, did not block the release of noradrenaline from the adrenergic nerve terminals of a nictitating membrane preparation, and did not alter the responses of smooth and cardiac muscle preparations to noradrenaline. It is suggested that the specificity of beta-bungarotoxin for certain nerve terminals is related either to selective binding of the toxin or to the selective presence of a necessary substrate for its action. An attempt to show selective binding of 125I-toxin to cholinergic nerve terminals in skeletal muscle was not successful.     

Influence of alpha and beta adrenergic receptors blockade on the adrenolytic effect of muscular work

The changes in the response of adrenergic receptors alpha and beta in the blood vessels in the working muscles in a hindlimb in cats were studied after intra-arterial administration of noradrenaline, isoprenaline and during electric stimulation of the sympathetic trunk. The experiments were carried out during alpha-adrenergic receptors blockade with dihydroergotamine (0.3 mg/kg) beta-adrenergic receptors blockade with propranolol (1 mg/kg) and blockade of acetylcholine M receptors with atropine (0.5 mg/kg). The investigations were performed at rest, during exercise (electric stimulation of the sciatic nerve) and after the exercise. The following results deserve attention: 1) beta-adrenergic receptors blockade reduced significantly the alpha-adrenolytic effect of exercise restoring the ability of blood vessel to constriction in response to noradrenaline; 2) the vasodilator effect of isoprenaline evident in resting state and maintained to some extent during exercise was abolished completely by preceding alpha-adrenergic blockade. The changes in the reactivity of resistance vessels in working skeletal muscles to noradrenaline, with abolition of its vasoconstrictor effect, have been shown by Rein [7] and others authors [2, 5]. Similarly, it is well known that the resistance vessels contain two types of adrenergic receptors alpha and beta, and that the response of the vessels to stimulation of these receptors are different [1]. In view of the recently published observations of Jarhult and Lundvall suggesting that the beta-adrenergic receptors play an important physiological role [6] in the arterial part of the microcirculation [6] and in view of the hypothesis put forward by Kunos and Szentivanyj that alpha and beta receptors can be transformed depending on the intensity of tissue metabolism [8] it seemed worth while to study more systematically the changes of the reactivity of alpha and beta adrenergic receptors in the vascular bed of the skeletal muscles during and after muscle exercise.     

Mechanisms responsible for the cardiotoxic effects of cocaine

Cocaine can induce lethal cardiovascular events, including myocardial infarction and ventricular fibrillation. The mechanisms responsible for these cardiotoxic effects of cocaine remain largely to be determined. Cocaine has both sympathomimetic (inhibition of neuronal uptake of norepinephrine) and local anesthetic (Na+ channel blockade) properties. Neurotransmitters released from cardiac sympathetic nerves bind to both alpha- and beta-adrenergic receptors eliciting a cascade of intracellular responses. Stimulation of beta-adrenergic receptors activates adenylate cyclase, increasing cyclic AMP levels, whereas alpha-adrenergic receptor stimulation activates phospholipase C, increasing inositol trisphosphate. These second messengers, in turn, elicit increases in cystolic calcium. Elevations in cystolic calcium can provoke oscillatory depolarizations of the cardiac membrane, triggering sustained action potential generation and extrasystoles. Cocaine also acts as a local anesthetic by inhibiting sodium influx into cardiac cells, which impairs impulse conduction and creates an ideal substrate for reentrant circuits. Thus, the adrenergic and anesthetic properties of cocaine could act synergistically to elicit and maintain ventricular fibrillation. Adrenergic receptor activation would trigger the event whereas sodium channel blockade would create the reentrant substrate to perpetuate the malignant arrhythmias.     

Uptake and accumulation in vitro of 3H-noradrenaline in adrenergic nerves of human atrium

Summary The adrenergic innervation and the in vitro uptake of ³H-noradrenaline has been investigated in human atrial tissue slices from patients undergoing thoracic surgery. The atrial appendage was richly innervated, but the density of the adrenergic nerve plexus varied considerably between different tissues examined. The nerve terminals were of characteristic varicose appearance, running singly or in bundles along the long axis of the muscle fibers. The nerve fibers seemed to penetrate in between the muscle cells. The distribution and appearance of the adrenergic nerves were quite similar to those described in earlier investigations of heart tissue from other species. The uptake and accumulation of ³H-noradrenaline in vitro increased with increasing concentration in the medium and with time, and the uptake process could efficiently be blocked by desmethylimipramine (DMI), a potent inhibitor of the uptake mechanism located at the axonal membrane, the so called membrane pump. There was a true accumulation of ³H-noradrenaline in the atrial tissue during the incubation, compared to the medium. The metabolism of ³H-noradrenaline during the incubation has also been studied. The data presented speak in favour of the view that the adrenergic nerves of human atria possess an efficient uptake accumulation mechanism for noradrenaline.Abbreviations used DMI desmethylimipramine - NA noradrenaline - NM normetanephrine     

豚鼠肠系膜下神经节细胞电活动与结肠运动的关系

在豚鼠肠系膜下神经节(IMG)及其支配的结肠段联合标本上,对IMG细胞内电位与肠段纵肌或环肌舒缩活动进行了同步记录。实验结果表明:(1)肠段预置张力为零时,约50％IMG细胞有自发的快兴奋性突触后电位(EPSP)活动,切断结肠神经或以筒箭毒(50μmol/L)灌流IMG后消失;(2)筒箭毒或低钙高镁溶液阻断神经节传递时,环肌节律性收缩幅度增大,节律变慢,但对纵肌节律性收缩无明显影响,(3)串刺激节前神经,在IMG细胞引起一串快EPSP或动作电位并常跟随迟慢的EPSP,同时,纵肌在0.1-0.2s潜伏期后出现迅速的、时程基本与动作电位串一致的舒张波,后者在筒箭毒灌流IMG后消失,而环肌运动可见舒张、舒张波延长或收缩波增大。结果提示:IMG不仅中继经典的胆碱能传出功能,还参与以胆碱能传递为中介的肠-肠反射,该反射活动的传出效应主要在于抑制环肌收缩。