Different effects of neuropeptide Y on electrically induced contractions in the longitudinal and circular smooth muscle layers of the female rabbit urethra

The effects of neuropeptide Y (NPY) on preparations of isolated longitudinal and circular smooth muscle from rabbit urethra were studied. In both types of muscle, electrically induced contractions and relaxations could be abolished by tetrodotoxin, (TTX). In the longitudinal muscle preparations the contraction was slightly reduced by prazosin, but markedly reduced by scopolamine and NPY. The NPY effect was not influenced by pretreatment with rauwolscine. Pretreatment with NPY had no effect on contractions induced by noradrenaline (NA) or carbachol and the peptide did not relax preparations contracted by these agents. In circular muscle an initial, fast response, not sensitive to prazosin or scopolamine was occasionally observed following electrical stimulation. A slow contraction component was regularly seen; this response was abolished by prazosin. Neuropeptide Y did not influence any of these responses. The preparations were concentration-dependently contracted by NA, whereas carbachol had no effect. Pretreatment with NPY did not affect contractions induced by NA, nor did the peptide relax NA-contracted preparations. In neither longitudinal nor circular muscle strips did NPY affect the electrically induced TTX sensitive relaxation of NA-contracted preparations. The results suggest that in the rabbit urethra NPY reduces contractions in the longitudinal muscle layer by selectively inhibiting the release of acetylcholine from cholinergic nerves. Neuropeptide Y did not appear to have any significant postjunctional effects nor to interfere with the release, or effects of NA or other transmitter agents. The physiological importance of the urethral effects of NPY remains to be established.     

Nerve-mediated excitation of the taenia of the guinea-pig caecum

1. A study was made of the responses of the isolated taenia of the guinea-pig caecum to stimulation of the intramural nerves.2. Four types of response are described, made up of a contraction and/or a relaxation occurring during stimulation and an after-contraction occurring when stimulation is stopped.3. A delayed relaxation which sometimes occurred at the end of stimulation is also described.4. Atropine usually abolished the contractions occurring during stimulation. After-contractions either appeared or were increased in amplitude in the presence of atropine, and delayed relaxations were abolished.5. The anti-cholinesterase drug neostigmine could convert the response during stimulation from a relaxation to a contraction. After-contractions were often abolished by neostigmine and delayed relaxations appeared.6. It is concluded that the contractions which occurred during stimulation were mediated by cholinergic nerves. The after-contraction appears to be a rebound phenomenon, following the hyperpolarization of the muscle caused by stimulation of the inhibitory nerves. It is suggested that the delayed relaxation is caused by the effects of persisting inhibitory transmitter substance on cells which do not undergo rebound excitation.     

Possible involvement of neuropeptide Y in sympathetic vascular control of canine skeletal muscle

Sympathetic nerve stimulation (2 min, 2 and 10 Hz) increased perfusion pressure in the blood perfused canine gracilis muscle in situ after pretreatment with atropine, desipramine and beta-adrenoceptor antagonists. This vasoconstriction was accompanied by clear-cut increases in the overflow of endogenous noradrenaline (NA) at both frequencies and, at 10 Hz but not at 2 Hz, also of neuropeptide Y-like immunoreactivity (NPY-LI). The irreversible alpha-adrenoceptor antagonist phenoxybenzamine enhanced the nerve stimulation induced overflows of NA and NPY-LI five- to eightfold and threefold, respectively. The fractional overflows of NA and NPY-LI per nerve impulse were similar in response to the high-frequency stimulation, indicating equimolar release in relation to the tissue contents of the respective neurotransmitter. The maximal vasoconstrictor response elicited by 10 Hz was reduced by about 50% following a dose of phenoxybenzamine which abolished the effect of exogenous NA and the remaining response was more long-lasting. Local i.a. infusion of NPY evoked long-lasting vasoconstriction in the presence of phenoxybenzamine, while the stable adenosine 5(1)-triphosphate (ATP) analogue alpha-beta-methylene ATP was without vascular effects. Locally infused NPY reduced the nerve stimulation evoked NA overflow by 31% (P less than 0.01) at 1 microM in arterial plasma, suggesting prejunctional inhibition of NA release. In conclusion, NPY-LI is released from the canine gracilis muscle upon sympathetic nerve stimulation at high frequencies. There is nerve stimulation evoked vasoconstriction, which is resistant to alpha-adrenoceptor blockade. This may in part be mediated by NPY released together with NA from the sympathetic vascular nerves.     

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)     

Cholinergic and adrenergic neural control of smooth muscle function in the non-pregnant rat uterine cervix

The isolated circular smooth muscle of the uterine cervix from spayed rats was investigated in vitro. One group of animals was treated with oestrogen. Preparations from these rats showed no spontaneous contractile activity, but responded with contractions to electrical field stimulation. The contractions were blocked by approximately 85% with atropine, scopolamine and tetrodotoxin, but were unaffected by the adrenergic antagonists propranolol, phenoxybenzamine, and yohimbine. This neurogenic effect was potentiated by neostigmine. Noradrenaline inhibited the nerve-induced contractions and also lowered resting tension in a concentration-dependent way. The latter effect of noradrenaline was inhibited by propranolol. The indirect effects of noradrenaline on the electrically induced contractions were also inhibited by propranolol, but potentiated with phenoxybenzamine and, to a less extent, yohimbine. Nerve-induced relaxation was never observed. Acetylcholine contracted the smooth muscle by an effect that was counteracted by atropine. A second group of spayed rats was not given any steroid treatment. The cervix preparations from these animals showed spontaneous activity, which was uninfluenced by atropine, propranolol, phenoxybenzamine, yohimbine, and tetrodotoxin. Noradrenaline inhibited the spontaneous activity. This noradrenergic effect was counteracted by propranolol and potentiated by phenobenzamine. Acetylcholine increased the smooth-muscle tone by an atropine-sensitive action. The results suggest the presence of a cholinergic motor innervation on the non-pregnant rat uterine cervix. The cervix also receives an adrenergic innervation, which may act both post-junctionally (lowering resting tension and inhibiting spontaneous activity) and prejunctionally (modulating the cholinergic nerves) via beta-adrenergic as well as alpha-adrenergic receptors.     

Nervous regulation of the tone of the retractor penis muscle in the goat

The nervous control of the retractor penis muscle (rp) was investigated in the anaesthetized goat. Also, isolated field stimulated strips of the muscle were studied. The noradrenaline (NA) and acetylcholine (ACh) content of the rp was determined, and histochemistry for adrenergic and acetylcholinesterase (AChE) positive nerves was performed. The muscle exhibited spontaneous activity that persisted after section of all nerves. There was, however, also a tendency of the activity to follow the general vasomotor tone, which disappeared after section of the sympathetic chains. The excitatory adrenergic nerves which innervate the muscle come from the sympathetic chains and run along the pudendal, the hypogastric and the pelvic nerves. The rp has a dense network of adrenergic fibres and is very sensitive to excitatory adrenergic stimulation. It has a fairly large NA content, which is higher in old goats (5.95 ± 0.42 μg g^-1) than in young goats (2.87 ± 0.78 μg g-¹). Inhibitory non-adrenergic non-cholinergic (NANC) innervation reaches it via the pelvic and the hypogastric nerves. The maximum inhibitory response is reached at low frequencies (2–4 Hz). Cholinergic prejunctional inhibition of the excitatory response to sympathetic chain stimulation was effected by simultaneous stimulation of the hypogastric nerves. In vitro experiments confirmed the presence of endogenous cholinergic muscarinic suppression of the excitatory adrenergic neurotransmission. Significant amounts of ACh (0.81 7 plusmn; 0.18 μg g^-1) are present in the muscle, and it contains strongly AChE positive nerve fibres and nerve cell bodies. It is concluded that the goat rp is innervated by sympathetic adrenergic excitatory nerves and parasympathetic NANC inhibitory nerves. It further has a direct sympathetic inhibitory NANC innervation, and an indirect inhibitory cholinergic innervation which at least in part is sympathetic.     

Effects of transmural field stimulation in isolated muscle strips from rabbit sphincter of Oddi and duodenum

The purpose of the study was to compare the effect of transmural field stimulation (TMS) on isolated smooth muscle strips from rabbit sphincter of Oddi (SO), duodenal circular layer (D_c) and duodenal longitudinal layer (D. The strips were suspended in thermostatically controlled 5-ml organ baths containing Krebs solution constantly bubbled with 5% CO_a in 0₂. TMS was delivered through platinum electrodes (140 V, 0.4 ms, 5 s trains, 40 Hz). The TMS responses could be divided in two main responses: (1) contraction initiated after cessation of the stimulus train, preceded by an inhibitory phase during TMS (‘off’); and (2) contraction initiated during TMS (‘duration’). The ‘duration’ response was observed in one out of 20 strips in the SO and D_c compartments, whereas 11 D, strips (55%) showed ‘duration’ responses (P < 0.001). Atropine (10^-6) converted all ‘duration’ responses to an ‘off’ response preceded by an inhibitory phase during TMS and reduced the contractile amplitudes with 40–65%. L-NNA significantly increased the number of ‘duration’ responses in all types of muscle, and caused a 40% increase in Dj contractile amplitude. Inhibitory responses could not be removed by atropine, propranolol and phentolamine. The results suggest that the intrinsic innervation of SO and duodenal muscle consists of a mixture of excitatory, cholinergic and inhibitory NANC pathways. The latter may utilize, wholly or partly, NO or a related compound as transmitter. A relative dominance of excitatory, cholinergic responses was present in the D_: strips, whereas inhibitory responses were dominating in the SO and D_c strips.     

Origin and peptide content of nerve fibers in the nasal mucosa of rats

Injection of the retrograde neuronal tracer True blue into the anterior-lateral part of the nasal mucosa of rats labeled nerve cell bodies in the superior cervical ganglion, the sphenopalatine ganglion, the otic ganglion and the trigeminal ganglion on the ipsilateral side. In the superior cervical ganglion, the sphenopalatine ganglion and the trigeminal ganglion on the contralateral side, very few nerve cell bodies were labeled, indicating that these ganglia provide minor contributions only. The number of labeled cell bodies indicates that the superior cervical ganglion, the sphenopalatine ganglion and the trigeminal ganglion contribute most to the innervation of the nose, while the contribution from the otic ganglion is minor. Cell bodies in the superior cervical ganglion harbored noradrenaline (NA) or NA/neuropeptide Y (NPY); in the sphenopalatine ganglion vasoactive intestinal peptide (VIP) or VIP/NPY; in the otic ganglion VIP, VIP/NPY or VIP/substance P (SP) and in the trigeminal ganglion calcitonin gene-related peptide (CGRP) or CGRP/SP. The results from denervations and tracer experiments suggest that all NA-containing and the majority of NPY-containing fibers in the nasal mucosa are derived from the superior cervical ganglion (sympathetic nerve supply). VIP- and VIP/NPY-containing fibers originate from the sphenopalatine and otic ganglia (parasympathetic nerve supply). Nerve fibers containing CGRP and CGRP/SP emanate from the trigeminal ganglion (sensory nerve supply).     

Reinnervation of Müller's smooth muscle by atypical sympathetic pathways following neonatal ganglionectomy in the rat: structural and functional investigations of enhanced neuroplasticity

Müller's extraocular smooth muscle is reinnervated by sympathetic nerves following denervation by ipsilateral superior cervical ganglionectomy in neonates but not in older animals. Experiments were performed to determine: (1) the source and extent of reinnervation, (2) the role of impulse activity in sympathetic outgrowth and (3) the effects of reinnervation on smooth muscle maturation. Müller's muscles were evaluated structurally (muscle volume, catecholamine histochemistry, retrograde labeling of sympathetic neurons) and functionally (contractile responses to electrical stimulation of postganglionic innervation and adrenoceptor agonist) in control preparations and in muscles following neonatal ipsilateral superior ganglionectomy, ipsilateral decentralization, ipsilateral superior ganglionectomy combined with contralateral decentralization of chemical (guanethidine) sympathectomy. Fluorescent tracer injections of muscles in adult control rats labeled cells in the ipsilateral superior (98%) and middle cervical ganglia. Acute ipsilateral superior ganglionectomy produced complete degeneration of sympathetic innervation of Müller's muscle in neonatal and adult rats. In preparations denervated neonatally and maintained chronically, muscles were reinnervated by neurons in both the contralateral superior and ipsilateral middle cervical ganglia. The total number of neurons reinnervating the muscle was one half that of controls. Sectional density of innervation was 45% of control. Electrical stimulation of postganglionic axons in the contralateral pathway produced muscle contractions with a prolonged time course. Reinnervation alleviated, in part, deficits in muscle volume and contraction which occurred following sustained denervation by chemical sympathectomy. Decentralization decreased ipsilateral muscle volume but did not affect numbers of neurons projecting to or nerve density within the muscle. Stimulation frequencies required to produce a 50% maximum contraction were reduced in these preparations. Decentralization of the contralateral ganglion did not impede sprouting into the denervated muscle, as nerve density and number of labeled cells were comparable to muscles reinnervated by contralateral ganglia with intact preganglionic innervation. However, maximum contraction to electrical stimulation was reduced. Comparisons with ipsilaterally decentralized muscles revealed that increased stimulation frequencies were required for 50% maximum contraction.(ABSTRACT TRUNCATED AT 400 WORDS)     

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.     

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.     

Transmission from vasoconstrictor and vasodilator nerves to single smooth muscle cells of the guinea-pig uterine artery

1. Previous studies have shown that perivascular nerve stimulation of the uterine artery of the guinea-pig evokes an adrenergic constrictor response and a dilator response with two components. The first of these, only present during pregnancy, is cholinergic. The second is non-cholinergic and is present at all times. Intracellular recording from single smooth muscle cells in isolated arterial segments has now been used to investigate the transmission processes associated with these responses.2. The mean resting membrane potential of the muscle cells was 60.7 mV in arteries from both virgin animals (range 50-68 mV) and from animals in late pregnancy (range 48-76 mV).3. Low frequency perivascular stimulation evoked excitatory junction potentials (EJPs) which reached a maximum amplitude of about 5 mV, lasted about 900-1000 msec, and showed facilitation at frequencies of stimulation of 0.1 Hz or above and summation at frequencies of stimulation of 1.2 Hz or above.4. These EJPs were abolished by exposure of the tissue to bretylium (2 x 10(-6) g/ml.). It is therefore concluded that the EJPs were due to transmission from adrenergic nerves.5. Perivascular stimulation at frequencies above 10 Hz evoked a depolarizing response which was often surmounted by a small (5 mV) local spike potential. Such depolarizing responses were associated only with localized contractions of the arterial muscle.6. In the presence of low extracellular K(+) concentrations, perivascular stimulation at frequencies above 10 Hz gave rise to a depolarizing response topped by an action potential of up to 50 mV amplitude, and more generalized contraction of the tissue than was seen in normal K(+) solution.7. After blockade of the adrenergic vasoconstrictor fibres, no response to perivascular stimulation was observed normally. However, following moderate depolarization of the membrane with noradrenaline, stimulation evoked a hyperpolarization of up to 6 mV in amplitude.8. This response showed no discrete junction potentials, had a latency of up to 2000 msec and was only observed with stimulation at frequencies of 2 Hz or greater. The response was obtained in both pregnant and non-pregnant animals, and was unaffected by hyoscine, but was abolished by cinchocaine.9. No changes in membrane potential attributable to transmission from cholinergic dilator nerves could be revealed in arteries from pregnant animals. Furthermore, high concentrations of acetylcholine had no polarizing effect on the muscle cells. It is suggested that the cholinergic dilator nerves may not act via changes in membrane potential.     

Some electrical properties of the rabbit anococcygeus muscle and a comparison of the effects of inhibitory nerve stimulation in the rat and rabbit

1. Simultaneous recordings of mechanical activity and membrane potential of individual smooth muscle cells have been made in the rabbit anococcygeus muscle and the effect of field stimulation on these examined.2. In the absence of tone the mean resting membrane potential was - 48 mV. In the stretched muscle spontaneous tone and rhythmic activity quite frequently appeared and this was associated with depolarization of the muscle cells.3. The response to field stimulation depended on the frequency of stimulation, the level of membrane potential and the presence of myogenic tone. The usual response to single pulses or low frequency stimulation was a hyperpolarization of up to 30 mV (mean 14+/-6.8 mV) after a latency of 185 msec and accompanied by muscle relaxation. Higher frequencies (over 8 Hz) produced an initial depolarization often with a spike potential and followed by hyperpolarization. The mechanical response in these instances was contraction or contraction followed by relaxation. At all frequencies rebound depolarization and an associated contraction followed the end of stimulation).4. Phentolamine (5x10(-6)M) and guanethidine (10(-6)M) blocked the initial depolarization and contraction but had no effect on hyperpolarization, muscle relaxation or rebound depolarization and contraction.5. The effect of field stimulation in the presence of guanethidine (4x10(-5)M) was re-examined in the rat anococcygeus. Single pulses were ineffective, repetitive stimulation produced muscle relaxation but no hyperpolarization comparable to the rabbit. Any oscillations in membrane potential were damped during field stimulation and sometimes a small hyperpolarization was produced with a maximum amplitude of 13 mV and a mean of 1.9+/-1.2 mV.6. The transmembrane potential at the peak of hyperpolarization in the rabbit was rarely more than -70 mV. Passive displacement of the membrane potential by current pulses altered the amplitude of the hyperpolarization and suggested that there was a reversal potential at between -80 and -90 mV.7. No change in input resistance could be measured during inhibitory nerve stimulation in either the rabbit or the rat but measurements based on electrotonic potentials indicated a reducation in membrane resistance, small in the rat but greater in the rabbit.8. These experiments suggest that in both species muscle relaxation is associated with an increase in ionic permeability and a move, at least in the rabbit muscle, towards an equilibrium potential of -80 to -90 mV. In view of the much smaller effect in the rat it is not clear whether this is the cause or at least the sole cause of the muscle relaxation.     

Effects of neuropeptide Y (NPY) on isolated guinea-pig heart

Neuropeptide Y (NPY) is present in nerve fibres throughout the mammalian heart. We have elucidated the effects of NPY on the isolated papillary muscle and heart (Langendorff) from the guinea-pig. The paced papillary muscle was studied with regard to duration of the action potential, peak force, maximum rate of force development, time to peak force, and time from peak force to half relaxation; all these parameters were identical whether or not NPY (5 X 10(-7) M) was present in the medium. When a stimulation with trains of pulses was superimposed, the paced papillary muscle exhibited enhanced contractions. This increase in contractility was not observed in the presence of the beta-adrenoceptor antagonist propranolol (10(-6) M) and was thus considered to be adrenergically mediated. The latter (adrenergic) response was markedly attenuated by NPY. Since NPY did not interfere with the response to exogenous noradrenaline (NA) it is suggested that the peptide exerts a pre-junctional inhibitory affect on adrenergic nerve-mediated positive inotropy. Neuropeptide Y did not influence the electrocardiogram from the spontaneously beating heart (Lagendorff), nor did the peptide modify the positive chronotropic effect of exogenously applied NA. In conclusion, the results indicate that NPY is without effect on the heart muscle proper but that the sympathetic terminals of the heart possess pre-junctional receptors for NPY (and/or related peptides) related to suppression of stimulated NA release.     

Autonomic nervous control of cervical secretion in the guinea-pig

Neurogenic influence on cervical secretion was studied in experiments performed on isolated guinea-pig cervix with intact adjacent nerves. The secretion was estimated as carbohydrate efflux. Experiments were undertaken at ovulation time when secretion is most prominent. Hypogastric nerve stimulation significantly increased carbohydrate release. This response was unaffected by phentolamine but inhibited by TTX, scopolamine and hexamethonium, as well as previous destruction of paracervical ganglia. Pelvic nerve stimulation did not affect cervical secretion. Carbachol as well as VIP induced a secretory response of the same extent as did hypogastric nerve stimulation. Scopolamine abolished the VIP-induced secretion. VIP and carbachol in combination had no additive effect. It is concluded that cervical glands of the guinea-pig uterus possess a cholinergic secretomotor innervation. The main pathway is the hypogastric nerve and neurons have a peripheral relay at paracervical ganglia. A more complex peripheral organization cannot be excluded.     

Adrenoceptor action of neurons of the superior cervical and caudal mesenteric sympathetic ganglia in cats

Experiments on the superior cervical and caudal mesenteric sympathetic ganglia of cats showed that dopamine (DA), like noradrenalin (NA) and adrenalin (A), inhibits cholinergic conduction. DA activity in the superior cervical ganglion is 2 and 3 times less respectively than NA and A activity, whereas in the caudal mesenteric ganglion, DA is 50 times more active than NA as regards ability to inhibit cholinergic conduction. The effects of DA and NA in the superior cervical ganglion are abolished by dihydroergotamine, phentolamine, and haloperidol, but not by tropaphen and chloropromazine. In the caudal mesenteric ganglion the inhibitory effect of NA is reduced by phentolamine, dihydroergotamine, and chlorpromazine but not by haloperidol. Conversely, haloperidol and chlorpromazine reduced the inhibitory effect of DA on cholinergic conduction in the caudal mesenteric ganglion, whereas phentolamine, dihydroergotamine, and deseryl were ineffective. It is postulated that the high level of development of the dopaminergic mechanism of inhibition of cholinergic conduction in the caudal mesenteric sympathetic ganglion may lie at the basis of DA-induced dilatation of mesenteric and renal blood vessels and the hypotensive action of DA.     

Adrenergic,cholinergic and VIP-ergic influence on gastric phasic motility in the rat

Bojö , L., Cassuto , J., Nellgård , P. & Jönsson , A. 1994. Adrenergic, cholinergic and VIP-ergic influence on gastric phasic motility in the rat. Acta Physiol Scand 150, 67–73. Received 30 December 1992, accepted 23 July 1993. ISSN 0001–6772. Department of Clinical Physiology, Central Hospital Karlstad, Department of Physiology, University of Goteborg and Department of Anaesthesiology, Central Hospital, Molndal, Sweden. We investigated the effects of adrenergic, cholinergic and vasoactive intestinal polypeptide (VlP)-ergic agonists and antagonists on the amplitude of gastric phasic contractions in the anaesthetized rat using a volumetric model. The amplitude of the phasic contractions was reduced significantly by atropine, hexamethonium or bilateral cervical vagotomy indicating that cholinergic neural activity involving both muscarinic and nicotinic receptors and intact vagal nerve function are integral parts of the control of basal gastric phasic motility. In contrast, neither selective α₁-, α₂- or non-selective β-blockers had any significant influence on the amplitude of the gastric contractions suggesting that adrenergic neurones are not tonically active in the maintenance of basal phasic motility in the stomach. The amplitude of the gastric phasic contractions was, however, significantly reduced by the α₁-agonist L-phenylephrine, the α₂-agonist clonidine and a close intraarterial injection of VIP (3 μkg^-1) but not be the selective β₁-agonist, prenalterol, or the β₂-agonist, salbutamol. These data suggest the presence of superimposed inhibitory control of phasic activity by VIP-ergic stimulation and by adrenergic neurones via α-receptor stimulation.     

Polar innervation of enteric non-cholinergic and non-adrenergic neurons in the isolated guinea-pig ileum

To study the polarity of the efferent pathway of the myenteric plexus, recordings were made of the mechanical activity of the longitudinal muscle of isolated guinea-pig ileal segments upon stimulation with an electrical field around the myenteric plexus contained within strips of longitudinal muscle (LM-MP) continuous with each end of ileal segment. The amplitude of the contractile response to stimulation of the anal LM-MP was always larger than that to the oral LM-MP. After cholinergic and adrenergic transmission was suppressed by atropine (10 microM) and guanethidine (1 microM), and the tone of the segment was enhanced by histamine (1 microM), the LM-MP stimulation produced non-cholinergic, non-adrenergic (NCNA) ascending contraction and NCNA descending relaxation. The NCNA contraction, but not the NCNA relaxation, was abolished or reduced by desensitization to substance P. The present results suggest that the NCNA innervation of the myenteric plexus participates in the polar effects observed in the guinea-pig ileum, that the NCNA excitatory response may be mediated at least in part by myenteric substance P neurons, and that the NCNA inhibitory response is mediated by non-adrenergic neurons.     

Frequency- and reserpine-dependent chemical coding of sympathetic transmission: differential release of noradrenaline and neuropeptide Y from pig spleen

The importance of impulse pattern and stimulation frequency for the release of noradrenaline (NA) and the coexisting peptide neuropeptide Y (NPY) in relation to vasoconstriction (perfusion-pressure increase) was studied in the blood-perfused pig spleen in vivo. Splenic nerve stimulation with intermittent bursts at high frequency (20 Hz) caused a several-fold larger release of NPY-like immunoreactivity (-LI) in relation to NA than a continuous stimulation at a low frequency (2 Hz), giving the same total number of impulses. alpha-Adrenoceptor blockade by phentolamine enhanced markedly both NA and NPY release, especially at low stimulation frequency, suggesting prejunctional adrenergic inhibition of release. Addition of propranolol unmasked a large remaining perfusion-pressure response to nerve stimulation. Reserpine treatment reduced the NA content of the spleen as well as the stimulation-evoked NA release by greater than 90%. However, the perfusion-pressure increase in response to nerve stimulation was well maintained. A marked increase in the stimulation-evoked release of NPY-LI occurred after reserpine. Adrenoceptor blockade after reserpine treatment reduced only slightly the perfusion-pressure response in parallel with a decline in NPY output. NPY caused an adrenoceptor-resistant perfusion-pressure increase at plasma concentrations that were in the same range as the maximal increase during nerve stimulations. In conclusion, the present data suggest a frequency-dependent, chemical coding of sympathetic transmission with preferential release of the classical transmitter NA at low, continuous frequencies and release of NPY, mainly at high frequencies. Reserpine treatment enhances markedly NPY release, which may explain why the functional response is largely intact in spite of adrenoceptor blockade and marked NA depletion.     

Urinary bladder and urethral responses to pelvic and hypogastric nerve stimulation and their relation to vasoactive intestinal polypeptide in the anaesthetized dog

The effects on the urinary bladder and urethra of pelvic and hypogastric nerve stimulation and their relation to vasoactive intestinal polypeptides (VIP) were investigated in the anaesthetized dog. Both pelvic and hypogastric nerve stimulation elicited a twofold increase in urinary bladder blood flow and a clear-cut increase in bladder venous effluent VIP concentration. Hypogastric nerve stimulation induced an initial, partly alpha-adrenergic and partly non-adrenergic, non-cholinergic, contraction of the urinary bladder followed by a relaxation. The urethra response was a maintained alpha-adrenergic contraction. Pelvic nerve stimulation elicited a bladder contraction with an initial non-cholinergic peak, whereafter the bladder pressure was maintained at a lower level, an effect which was mainly cholinergic in origin. The urethral response was an initial non-adrenergic, non-cholinergic contraction followed by a maintained cholinergic contractile response. Afferent pelvic nerve stimulation led to an efferent activity that seemed to be a combination of activity in pelvic and hypogastric pathways to the urinary bladder and the urethra. VIP (10 nmol) injected i.v. induced a relaxation of the urinary bladder and the urethra, together with a fall in systemic blood pressure. However, despite high plasma concentrations, no vasodilation was elicited in the urinary bladder. Thus, the main target for the VIP release during pelvic and hypogastric nerve stimulation is probably not the bladder vasculature, but instead perhaps the bladder smooth muscle proper.