Evidences for alpha excitatory action of catecholamines on the electrical activity of the guinea-pig stomach.

Effects of catecholamines and their inhibitors on the electrical activity of the smooth muscle of the guinea-pig stomach were studied by using pressure electrodes. The spontaneous electrical activity of antrum and corpus preparations consisted of slow waves with or without superimposed spikes. In quiescent fundus preparations, the spike activity and slow waves were generated by TEA. Catecholamines (noradrenaline, isoprenaline and adrenaline) suppressed or blocked the spike discharges and the generation of slow waves. However, the spike activity was enhanced at a higher concentration of phenylephrine in the antrumand fundus preparations. Inhibitory responses of the smooth muscles from whole regions to adrenaline, noradrenaline or isoprenaline were antagonized by propranolol or DCI but not by phentolamine or phenoxybenzamine. Therefore, inhibitory actions of these amines appear to involve beta-adrenoceptors. After treatment with tetrodotoxin, noradrenaline and isoprenaline blocked the spike activity and the generation of slow waves. Phenylephrine or adrenaline potentiated the spike activity in the presence of tetrodotoxin. After treatment with DCI or propranolol, phenylephrine potentiated the spike activity of the antrum and fundus preparations. These excitatory effects were antagonized by phentolamine or phenoxybenzamine. It is concluded that excitatory actions of these amines are mediated by alpha-adrenoceptors rather than via a nervous pathway.     

Atropine-sensitive, tetrodotoxin-resistant contraction induced by noradrenaline in isolated cat rectum

Effects of noradrenaline (NA) on the isolated rectal circular muscle of the cats were studied in comparison with the effects on the internal anal sphincter (IAS). NA (10(-8)-10(-7) g/ml) caused tonic contraction in four of 15 strips of the rectum taken from 15 animals, and in all 15 strips of the IAS. Phenylephrine also induced rectal and IAS contraction. Rectal contraction induced by NA was resistant to phentolamine, yohimbine, propranolol, hexamethonium and tetrodotoxin, but blocked by atropine. IAS contraction induced by NA was resistant to propranolol, atropine, hexamethonium and tetrodotoxin, but blocked by phentolamine and yohimbine. It is suggested that an atropine-sensitive excitatory adrenergic mechanism other than the excitatory alpha-adrenergic mechanism exists in the rectal circular muscle.     

Autonomic Nerve Control of the Swimbladder of the Goldsinny Wrasse,Ctenolabrus rupestris

The autonomic nerve control of the swimbladder of the goldsinny wrasse has been studied by experiments with isolated strips from the muscularis mucosae, drug effects on gas secretion in vivo, fluorescent histochemistry and quantitative analysis of catecholamine content. Vagotomy, atropine and mecamylamine significantly inhibited induced gas secretion, suggesting a cholinergic vagal secretory innervation of the gas gland. The anterior part of the mucosa (secretory part) is contracted by acetylcholine and α-adrenoceptor agonists. These responses are competitively antagonized by atropine and phentolamine or yohimbine respectively. The posterior (resorbent) part of the mucosa is contracted by acetylcholine, but β-adrenoceptor agonists relax the acetylcholine-precontracted preparations. These responses are competitively blocked by atropine and propranolol respectively. The pA₂-values from all experiments with antagonists are well comparable with values obtained for the same drugs in other teleost or mammalian tissues. Tyramine, 5-hydroxytryptamine or phenylephrine had mixed effects on the posterior part. This effect is at least in part due to release of nervously stored catecholamines. Fluorescent histochemistry revealed a strong innervation by both smooth and varicose adrenergic fibres in all parts of the swimbladder, noradrenaline being the dominant catecholamine. Yellow fluorescent cells, which may contain 5-hydroxytryptamine, were also seen in the swimbladder mucosa. No conclusive evidence for a cholinergic innervation of the muscularis mucosae was obtained.     

Human ejaculatory duct: parameters of smooth muscle motor activity and modulatory role of autonomic drugs

The contractile behaviour and effects of several autonomic drugs on the motor activity of human isolated ejaculatory ducts were investigated. Ejaculatory ducts exhibited spontaneous contractions characterised by an amplitude of 2.35 +/- 0.28 mN, a duration of 62. 9 +/- 3.72 s and a frequency of 0.64 +/- 0.014 waves min-1. Acetylcholine (10-5-10-4 m) induced a slight increase in basal tone and in the frequency of the contraction waves. These effects were suppressed by atropine (10-4 m). Noradrenaline (norepinephrine) increased the basal tone and frequency of spontaneous contractions in a dose-dependent manner. These responses were competitively inhibited by HEAT, a selective a1-adrenoceptor antagonist. These preliminary functional findings, indicating the presence of spontaneous motor activity of human ejaculatory ducts and its possible control by adrenergic agonists, suggests a physiological role for human ejaculatory duct in the propulsion of semen from the seminal vesicle towards the urethra.     

Presynaptic inhibitory actions of adenine nucleotides and adenosine on neurotransmission in the rat vas deferens.

Adenine nucleotides and adenosine inhibited the isometric contractions of the rat vas deferens in vitro in response to field stimulation but had no effect on the responses to exogenous noradrenaline. The inhibitions were potentiated by dipyridamole and compound 555, antagonized by theophylline and unchanged by indomethacin, 2-2′-pyridylistogen, phenoxybenzamine and atropine. Adenosine and adenosine 5′-triphosphate inhibited the release of [³H]noradrenaline produced by field stimulation.These results indicate that adenine nucleotides, probably acting via the common metabolite adenosine, inhibit adrenergic neurotransmission at a presynaptic site. Their antagonism by theophylline suggests that a presynaptic ‘purinergic’ receptor system could be involved.     

The long-lasting contraction induced by transmural stimulation in the longitudinal muscle of the guinea-pig gastric corpus

The mechanisms involved in the long-lasting contraction induced by transmural stimulation were investigated in the isolated longitudinal muscle of the guinea-pig gastric corpus. Transmural stimulation induced an initial rapid contraction, a subsequent relaxation and a long-lasting contraction which was mimicked closely by the responses to noradrenaline and ATP. The rapid contraction was blocked by tetrodotoxin or by atropine, and the subsequent relaxation was also blocked by tetrodotoxin, but not by atropine. The long-lasting contractions were not inhibited by atropine, phentolamine, and propranolol. However, some of those were partially inhibited by tetrodotoxin and guanethidine, and others were enhanced by tetrodotoxin. The noradrenaline- but not ATP-induced long-lasting contraction was blocked by phentolamine. Furthermore, indomethacin completely abolished the long-lasting contractions induced by transmural stimulation, noradrenaline, and ATP, but did not significantly affect the response to prostaglandin E2. The results indicate that the long-lasting contraction induced by transmural stimulation is mediated through prostaglandins release which may be triggered by neuro-transmitters or by endogenous substances.     

Agonist-induced secretions and potassium release from rat submandibular gland slices.

Secretory activity induced by stimulation of alpha-adrenergic, beta-adrenergic, and muscarinic cholingeric receptors and by dibutyryl cAMP and 8-bromo cGMP has been investigated in rat submandibular tissue slices. Isoproterenol produced a sialic acid secretion from the acinar cells that was inhibited by propranolol, but not by phenoxybenzamine or atropine. Dibutyryl cAMP produced a sialic acid secretion from the acinar cells that was not inhibited by propranolol, phenoxybenzamine, or atropine. Both norepinephrine and acetylcholine produced significant secretion of sialic acid but at a reduced efficacy. Norepinephrine stimulated both peptide hydrolase secretion from the granular duct cells and a release of K+ from the acinar cells. Both actions were inhibited by phenoxybenzamine, but not by propranolol or atropine. Acetylcholine stimulated a minimal secretion of peptide hydrolase from the granular duct cells and a significant release of K+ from the acinar cells. The norepinephrine- and acetylcholine-stimulated release of K+ was increased after the addition of 1 mM ouabain. High concentrations of 8-bromo cGMP induced a K+ efflux that was not inhibited by phenoxybenzamine or atropine. Vacuolation of the acinar cells was correlated with K+ release.     

Structural and functional ontogenetic development of the rat portal vein after neonatal 6-hydroxydopamine treatment

The importance of the adrenergic vasomotor nerve supply for the vascular ontogenetic development has been studied in the isolated portal vein preparation from rats, at 5–6 weeks of age, who had either been chemically sympathectomized by a series of postnatal 6-hydroxydopamine (6-OHDA) injections or been receiving the solvent alone. It was found that 6-OHDA treatment largely, but not completely, prevented the outgrowth of the terminal NA fluorescent ground plexus. Nevertheless, the media underwent a seemingly normal differentiation into two layers. Functionally, the portal vein from the 6-OHDA treated animals displayed weak and non-persistent myogenic spontaneous activity; sensitivity to exogenous noradrenaline (NA) was increased 3-fold and maximum stress was increased by 25 % as compared to control. Responses to transmural field stimulation were only obtained at high impulse rates and the maximum response was attenuated. Considering the very sparse adrenergic innervation following 6-OHDA they seemed surprisingly large, however, but since they were abolished by tetrodotoxin and by phenoxybenzamine responses are concluded to be neurogenic and adrenergic in origin. A singular attenuation of neurogenic responses by atropine was found in 6-OHDA treated vessels but not in controls. It is concluded that the adrenergic vasomotor nerve supply seems to exert some trophic influence during ontogenetic development but that the morphologic vascular development is largely governed by other, non-neurogenic mechanisms. As to functional development, 6-OHDA induced sympathectomy causes impaired development of phasic myogenic activity whereas maximum stress is augmented as is the tissue sensitivity to exogenous NA.     

Effects of some autonomic drugs on duodenal smooth muscle.

Longitudinal muscle strips (LMS) and circular muscle strips (CMS), 2 mm wide and 1.5--2 cm long, from opossum duodenum were exposed to some autonomic agonists. The cholinergic agonists, acetylcholine, carbachol, methacholine, and bethanechol stimulated only tonic contractions in LMS and tonic followed by phasic contractions in CMS. These effects were abolished by atropine 10(-6) M. The ED50S of all cholinergic agonists for LMS were significantly lower than for CMS. Norepinephrine caused initial contraction (abolished by phenoxybenzamine, 10(-4) M), followed by relaxation (abolished by propranolol, 10(-5) M), and isopropylnorepinephrine caused relaxation (abolished by propranolol, 10(-5) M) in both layers. There were no differences in relative potencies for adrenergic agonists between the layers. Tetrodotoxin did not affect the response to adrenergic agonists. Thus, the potency of cholinergic agonists is greater in longitudinal than in circular muscle, and the layers respond differently to cholinergic agonists. The alpha-adrenergic receptors mediate contraction and beta-adrenergic receptors mediate relaxation on the duodenal smooth muscle.     

Inhibition of the cholinergic neurotransmission in human airways via prejunctional alpha-2-adrenoceptors

Previous investigations on guinea-pig airways indicate that the excitatory, cholinergic neurotransmission can be inhibited via prejunctional alpha-2-adrenoceptors. The aim of this study was to investigate whether a similar inhibition of the cholinergic neurotransmission is present in human airways. Ring preparations of human bronchi were mounted for recording of isometric tension and immersed in oxygenated Krebs solution. Electrical field stimulation of the preparations elicited atropine-sensitive twitch contractions. Exogenous noradrenaline (in the presence of cocaine and propranolol) inhibited the electrically evoked contractions. The noradrenaline-induced inhibition could be antagonized by yohimbine whereas prazosin was ineffective, indicating that the inhibition was mediated by alpha-2-adrenoceptors. Contractions evoked by exogenous acetylcholine (in the presence of cocaine and propranolol) was unaffected by the addition of noradrenaline, which suggests that the alpha-2-adrenoceptors have a prejunctional localization. In conclusion, this report gives evidence that the human bronchial, cholinergic neurotransmission can be inhibited by stimulation of prejunctional alpha-2-adrenoceptors.     

Inhibitory effect of midaglizole on alpha adrenoceptor agonist-induced contractions of canine tracheal smooth muscle

To determine why midaglizole is effective in some patients with severe asthma, we investigated the inhibitory effects of midaglizole, prazosin or yohimbine on the BHT 920-, phenylephrine-, or noradrenaline-induced contractions of canine tracheal smooth muscle. After pretreatment with atropine (10(-6) M) and propranolol (10(-6) M) and precontraction with serotonin (3 x 10(-7) M), the tracheal muscle showed contractile responses to the exogenous administration of both alpha 1 and alpha 2 adrenoceptor agonists. Every alpha antagonist inhibited these agonist-induced contractions. Inhibitory activity of midaglizole (10(-4) M) for the alpha agonists was BHT-920 greater than noradrenaline greater than or equal to phenylephrine, while that of prazosin (3 x 10(-6) M) was phenylephrine greater than noradrenaline greater than BHT-920. Moreover, yohimbine completely inhibited the contractions at the lower concentration of 3 x 10(-7) M than that of other two antagonists. Our findings demonstrate that midaglizole dose-dependently inhibits airway contractions induced by alpha adrenoceptor agonists.     

Influence of intrinsic nerves on electromyogram of cat colon in vitro

The electromyogram of the circular muscle layer of the cat colon was studied in vitro in superfused strips of muscle. Records exhibited electrical slow waves and migrating spike bursts, as described previously. Both the neurotoxin, tetrodotoxin, and the local anesthetic lidocaine, (P less than 0.05) prolonged the duration of migrating spike bursts, but migrating spike bursts were not affected by the adrenergic alpha-antagonist, phenoxybenzamine, nor by the adrenergic beta-antagonist, propranolol. Also, physostigmine and atropine did not affect them. Large concentrations of catecholamines did abolish them. This suggests that the migrating spike burst represents periodic release of the muscle from the tonic influence of nonadrenergic inhibitory nerves in the intramural plexuses. Slow-wave frequency and the congruence of slow waves were not affected (P greater than 0.05) by the antagonists listed above, nor by cholinergic and adrenergic agonists. This suggests that the slow waves are not importantly controlled by intrinsic nerves.     

Pharmacological analysis of the mechanism of action for colonic contraction induced by neurotensin, substance P and methionine-enkephalin

Infusion of neurotensin, substance P and methionine-enkephalin induces colonic contraction in the cat. The present study was performed to investigate the effect of various pharmacological blocking agents on colonic contraction evoked by these peptides infused by the i.a. or i.v. route. The contractions caused by infusions of neurotensin were blocked by tetrodotoxin (1 micrograms kg-1 i.a.), hexamethionium (10 mg kg-1 i.v.), atropine (0.1 mg kg-1 i.v.) or somatostatin (100 pmol min-1 i.a.), but not by haloperidol, methysergide, mepyramine, cimetidine or naloxone. The contractile effect of substance P on the colon was abolished by the substance P receptor antagonist (D-Arg1, D-Pro2, D-Trp7,9, Leu11)-substance P (70 nmol min-1 i.a.). No other blockers used, such as tetrodotoxin, hexamethonium, atropine, mepyramine, cimetidine, methysergide, naloxone or somatostatin inhibited the response to substance P. Methionine-enkephalin produced a colonic contraction that was completely blocked by naloxone (1 mg kg-1 i.a.). Both atropine (0.1 mg kg-1 i.v.) and somatostatin (100 pmol min-1 i.a.) reduced the contractile response. However, tetrodotoxin, hexamethonium, mepyramine, cimetidine and methysergide did not affect the response to methionine-enkephalin. All adrenergic blockers tested, that is, guanethidine, propranolol and phentolamine, increased the contractile responses to the peptides. The results indicate that the colonic contraction induced by neurotensin is mediated via nervous cholinergic pathways. Substance P induces colonic contraction, probably by a direct effect on smooth muscle substance P receptors. Methionine-enkephalin induces colonic contraction which could be blocked by naloxone. However, a cholinergic or peptidergic link may also be involved in the response to methionine-enkephalin.     

A reciprocal adrenergic-cholinergic axoaxonic synapse in the mammalian gut.

Simultaneous stimulation of perivascular nerves inhibited the release of acetylcholine from stimulated cholinergic nerves of the rabbit jejunum. Adrenergic nerves were responsible for this inhibition because it did not occur in animals previously injected with 6-hydroxydopamine. Acetylcholine inhibited the release of transmitter from stimulated adrenergic axons; this effect was blocked by atropine. Since atropine enhanced the release of adrenergic transmitter when both adrenergic and cholinergic nerves were activated simultaneously (at 4.0 Hz), it seems likely that cholinergic nerves also inhibit release of norepinephrine (NE). Radioautographic examination of the myenteric plexus, incubated with tritiated NE, revealed a striking marginal distribution of adrenergic axons around the periphery of the myenteric plexus. Ultrastructural studies, with KMnO4 used to identify adrenergic terminal varicosities, confirmed this distribution and also revealed complexes formed between the terminal varicosities of adrenergic and probable cholinergic axons. The component varicosities forming these complexes contacted one another with no intervening Schwann cell elements. It is concluded that there is a reciprocal axoaxonic synapse between adrenergic and cholinergic neurons in the mammalian myenteric plexus.     

On a cholinergic motor innervation of the rat urethra

Choline acetyltransferase activity was demonstrated in the proximal part of the male rat urethra, indicating a cholinergic innervation of this tissue. The cholinergic nerve fibres emanated evidently from the pelvic nerves, since bilateral removal of the pelvic ganglion caused a major fall in the activity of this enzyme. The muscle activity of the circular layer of the proximal urethra was recorded in vitro. The basal activity of this segment was low. The parasympathomimetics acetylcholine and methacholine, evoked rapid and marked contractile responses; the maximal responses to these drugs were 36 and 44%, respectively, of that to potassium. The corresponding figures for phenylephrine and noradrenaline were found to be 79 and 88%, respectively. The responses evoked by the parasympathomimetics were unaffected by the ganglion blocker hexamethonium, the alpha-adrenoceptor blocker dihydroergotamine and the beta-adrenoceptor blocker propranolol. Atropine, however, abolished the responses completely. Following degeneration of adrenergic or cholinergic nerves of the urethra the parasympathomimetics still evoked contractions. Taken together these findings indicate that the parasympathomimetics exert their contractile effect through a direct action on muscarinic receptors. Parasympathectomy but not sympathectomy (caused by 6-hydroxydopamine treatment) gave rise to a supersensitivity to methacholine, as judged by a leftward shift of the dose-response curve for this drug, the ED50-value being ten times less than that of the controls. The observations seem to suggest that the proximal urethra normally is under the influence of cholinergic activity beside that of adrenergic activity previously demonstrated.     

Spontaneous activity in isolated bovine mesenteric lymphatics

1. Longitudinal tension was measured in isolated segments (approximately 20 mm in length and 2 mm in diameter) of bovine mesenteric lymphatic vessels.2. Seventeen out of thirty-nine vessels showed regular spontaneous rhythmic contractions, the most common frequency being about 2/min.3. The strength of each contraction was related to the duration of the preceding pause, a longer pause being associated with a stronger contraction.4. Noradrenaline increased the frequency and decreased the amplitude of the contractions. This effect was abolished by alpha-receptor blockade with phentolamine.5. Isoprenaline slowed the frequency of the contractions. This effect was abolished by beta-receptor blockade with propranolol.6. Acetylcholine had little effect on the rate of spontaneous contractions.     

Autonomic nervous influence of the female guinea-pig urinary bladder during the oestrus cycle

The mammalian urinary bladder receives dual innervation. The excitatory innervation is considered to be partly cholinergic and partly mediated via NANC-receptors. Several (co-)transmitters have been suggested. The adrenergic inhibitory innervation is mediated via α- and β-receptors. Female sex hormones could change autonomic influence of urogenital organs. It was considered to be of interest to characterize the spontaneous and nerve stimulation-induced muscular activity in the urinary bladder of the female guinea-pig during the oestrus cycle. Both the spontaneous activity and nerve-induced activity varied according to the hormonal status of the animal. An α-adrenergic inhibitory influence was identified. It was further confirmed that the excitatory innervation could not be blocked by the cholinergic antagonist scopolamine, while α-β-methylene ATP partly inhibited nerve stimulation-induced smooth muscle response, most prominent at cycle day 6. Indomethacin did not impair spontaneous activity or nerve stimulation-induced activity. Nitric oxide reduced nerve stimulation-induced responses on cycle day 12. Imperative urinary bladder contractions are reported to diminish after oestrogen use and in the female a hormonal effect of the nervous influence on the urinary bladder smooth muscle is suggested.     

Prolonged exposure to NT-3 attenuates cholinergic nerve-mediated contractions in cultured murine airways

Chronic airway inflammation may induce subsequent airway hyper-responsiveness (AHR) including pathological alteration of neural activity. Asthmatic airways contain elevated levels of neurotrophin-3 (NT-3) and brain-derived neurotrophic factor (BDNF) albeit, their effect on neural activity is unclear. This study evaluates the effects of NT-3 and BDNF on nerve mediated airway contractions in vitro. Tracheal segments from BALB/c J mice were cultured for 4 days with NT-3 or BDNF. Responsiveness to electric field stimulation (EFS) was evaluated in organ-bath and innervation patterns were examined by quantitative immunohistochemistry. In cultured segments the EFS-induced contractions were inhibited by tetrodotoxin or atropine. NT-3 reduced the EFS contractions in a concentration-dependent manner whereas BDNF had no effect. The amount of nerve fibers, found in conjunction with the tracheal smooth muscle, was similar in NT-3 treated and control segments. In conclusion, NT-3 attenuates cholinergic nerve-mediated contractions of airway in vitro. Considering the elevated levels of NT-3 found in asthmatic airways, the findings imply a protective role of NT-3 in AHR.     

Transmitter characteristics of small mesenteric arteries from the rat

We have studied the neurogenic response of small mesenteric arteries from the rat to evaluate the involvement of possible co-transmitters under various modes of stimulation. Segments of small branches of the mesenteric artery were mounted in a myograph and the intramural nerves were activated with transmural electrical stimulation. A single stimulation of the nerves caused a contraction that was reduced by only 20% in the presence of adrenergic blocking agents (prazosin or phenoxybenzamine), whereas the steady-state response to continuous nerve stimulation of high frequency was reduced by 90-95%. In contrast, all responses to applied noradrenaline in doses up to at least 1 mM were eliminated by phenoxybenzamine treatment. The stable ATP analogue, alpha,beta-methylene ATP, reduced the response to a single nerve stimulation by 70%, but reduced the contraction caused by continuous high-frequency nerve stimulation by only 10%. None of these agents affected the response to applied neuropeptide Y (NPY). The response of relaxed vessels to nerve stimulation was totally blocked by the combination of an adrenoceptor-blocking agent and alpha,beta-methylene ATP, although even in this situation a further neurogenic response could be revealed in vessels precontracted with vasopressin. Responses to either single stimuli or brief burst stimulations were potentiated after high-frequency stimulation. Both the adrenergic and non-adrenergic components were enhanced to roughly the same extent. Also the potentiated response was eliminated by the combined application of prazosin and alpha,beta-methylene ATP. The non-adrenergic transmitter in the sympathetic nerves of small arteries thus appears to be the dominant transmitter during low-frequency nerve stimulation, causing rapid but phasic activation. Noradrenaline is the most important transmitter for higher frequencies, exerting slower but sustained contractions. The post-stimulatory potentiation affects both the adrenergic and the non-adrenergic part of the neurogenic response.     

Evidence against adrenergic motor transmission in the guinea-pig vas deferens

1. Field stimulation of desheathed preparations of guinea-pig vas deferens, treated with a ganglion-blocking agent, has revealed the presence of two tetrodotoxin-susceptible components in the motor response, suggesting the existence of two sets of post-ganglionic motor nerve fibres of different excitability: one set responding maximally to pulses of 0.1-0.4 msec; the other, to pulses of 2 msec. No distinction could be made pharmacologically between the two components.2. Cooling potentiated that component in the twitch-responses which was due to stimulation of the more excitable fibres.3. The sensitivity of the longitudinal muscle to the motor action of noradrenaline was low and was subject to considerable animal variation. But normal responses to post-ganglionic field stimulation were elicited in noradrenaline-insensitive preparations, in which the twitches elicited by 5 pulses could not be matched with noradrenaline, even 100-125 mug/ml.4. In some forty experiments, small doses of noradrenaline inhibited the twitch-responses evoked by either set of motor fibres. This inhibition differed from that produced by isoprenaline in two respects. Firstly, propranolol did not antagonize the noradrenaline inhibition, thus excluding an action on beta-adrenoceptors; and secondly, noradrenaline did not depress contractions elicited by muscarine or by 5-methylfurmethide.5. Phenoxybenzamine, 10(-6) g/ml., produced a thousandfold reduction in the sensitivity of the muscle to the motor action of noradrenaline, without any decrease in the height of the twitches elicited by 0.1 or 1 msec pulses.6. The twitch-responses were not affected by combined alpha + beta adrenoceptor blockade with phentolamine and propranolol.7. Tyramine, amphetamine, tranylcypromine and prostaglandin E(2) inhibited the twitches but potentiated the contractile effect of noradrenaline.8. The twitch-responses and their inhibition by noradrenaline were present in preparations taken from reserpinized animals.9. Although the twitch-responses could be paralysed by bretylium or guanethidine, the foregoing results excluded adrenergic transmission at the motor endings. Cholinergic transmission was also excluded by negative findings with anticholinesterases, atropine, nicotine and (+)-tubocurarine.10. Motor transmission by histamine, 5-hydroxytryptamine, gamma-aminobutyric acid or ATP was also excluded.