Central and local actions of opioids upon reticulo-ruminal motility in sheep

The effects of opioids and naloxone on cyclical forestomach motility were determined in anaesthetized and conscious sheep. To assess central or peripheral opioid actions, differential routes of administration were used. Possible dynamic effects along the innervating vagovagal reflex arc were investigated electrophysiologically at the cervical level of the vagus nerve. Further, direct influences on the smooth muscle were evaluatedin vitro on isolated longitudinal reticular strips. Additionally, the effects of some spasmogenic agents were studied for comparative purposes. In anaesthetized sheep, opioids depressed in an identical manner both the amplitude of spontaneous cyclical contractions and contractions evoked by electrical stimulation of the distal end of the cut cervical vagus. In conscious sheep, low doses of normorphine and loperamide inhibited frequency and amplitude centrally (20 g/kg and 4 g/kg via carotid artery respectively), whereas locally higher dose levels (200 g/kg and 10 g/kg via coeliac artery respectively) affected only the amplitude of cyclical contractions. Furthermore the opioid peptides Leu-, Met-enkephalin and [D-Ala²-Met⁵]-enkephalinamide preferentially depressed the amplitude of cyclical motility most efficiently if administrated via the coeliac artery. These results indicate the presence both of a central opioid action depressing frequency and amplitude and of a local opioid action depressing only the amplitude of cyclical reticulo-ruminal motility. Opioids did not alter the resting discharge of afferent tension units and similarly failed to modulate tone of reticular stripsin vitro, suggesting that the opioids act locally on the intramural neuronal plexus, possibly by diminishing the output of excitatory transmitter. Whether substance P could play a role as a vagal excitatory transmitter besides the classically implicated acetylcholine has been discussed. The central opioid mechanism is probably not situated within the gastric centres but elsewhere in the brain. Naloxone ( 100 g/kg, jugular vein) stimulated the frequency of cyclical ruminal motility only in well-defined experimental conditions, probably via a central mechanism.     

Loperamide: evidence for a centrally mediated opioid effect on rumen motility in conscious goats and sheep

Loperamide inhibited the frequency and amplitude of cyclical contractions of the rumen in conscious goats (100 micrograms/kg, i.v. and sheep (250 micrograms/kg, i.v.). In goats, the inhibitory effect of loperamide could be prevented by pretreatment with the opiate antagonist naltrexone (greater than or equal to 12.5 micrograms/kg, i.v.) but not by pretreatment with the dopaminergic antagonist domperidone (500 micrograms/kg, i.v.). Intracerebroventricular administration of 1 microgram/kg loperamide in goats significantly depressed ruminal contraction frequency, whereas intravenous administration of 10 micrograms/kg loperamide did not affect cyclical motility. Administered via the carotid artery, loperamide (4 micrograms/kg) depressed both frequency and amplitude of cyclical contractions of reticulum and rumen, whereas the same dose was ineffective via the coeliac artery. In vitro, loperamide (10 nM-100 microM) had no influence on spontaneous activity or tone of the reticular longitudinal muscle strips. It is concluded that loperamide inhibits cyclical ruminal contractions through a central opioid pathway.     

Dopamine-sensitive receptors that evoke rumination and modify reticulo-ruminal activity in sheep

Dopamine (20 micrograms/kg) evoked rumination in sheep when injected as a bolus into the coeliac artery or into the left gastric artery but not when injected into the carotid artery. A mixed alpha-adrenoreceptor antagonist (phentolamine) and an alpha 2-adrenoreceptor antagonist (yohimbine) prevented dopamine from evoking rumination, but a dopaminergic antagonist (metoclopramide) did not. These findings suggest that dopamine stimulated rumination by acting upon alpha 2-adrenoreceptors situated in the area supplied by the left gastric artery, whereas dopamine injected intracerebrally may have evoked rumination by an alpha 2-adrenoreceptor effect in the central nervous system (Bueno et al., 1983) and the actions of intrajugular dopamine were exclusively upon peripheral adrenoreceptors located specifically in the gastric area. Dopamine (1 microgram/kg/min) infused into the carotid artery reduced the frequency of reticular contractions by acting upon a centrally located dopaminergic receptor mechanism sensitive to metoclopramide but not to phentolamine. When dopamine was infused into the coeliac artery or into the left gastric artery, the amplitude of reticular contractions was reduced by a peripheral mechanism sensitive both to metoclopramide and to phentolamine. Dopamine also reduced the amplitude of reticular contractions when infused into the carotid artery but to a lesser degree than when given into the coeliac or left gastric artery.     

Modification by 6-hydroxydopamine (6-OHDA) of the inhibition of extrinsic rumen contractions in sheep produced by beta-endorphin

In sheep, beta-endorphin (1 and 2 micrograms/kg) administered into the third cerebral ventricle caused a significant inhibition of the frequency of rumen contractions. The amplitude of the first rumen contractions, following immediately after the end of infusion, and the average amplitude of primary rumen contractions, were inhibited. Beta-endorphin caused general psychomotor excitability. These results suggest that an inhibitory mu and delta opioid system is involved in the control of forestomach motility and general behaviour in sheep. All effects of beta-endorphin were completely prevented by i.c.v. 6-hydroxydopamine (6-OHDA, 18.2 micrograms/kg) pre-treatment. These results suggest that beta-endorphin-induced inhibition of rumen motility is due to central noradrenergic system activation. The exact location of this noradrenergic system remains to be determined.     

Review: reticuloruminal motility--a pharmacological target with a difference?

The vagal sensory inputs to and motor outputs from the hindbrain gastric centres required for reticuloruminal motility were sampled directly in anaesthetized sheep using electrophysiological 'single fibre' techniques and indirectly in conscious, surgically prepared sheep. Drugs were administered by close-arterial injection into a carotid artery to observe central effects and into the coeliac artery to observe peripheral effects on the reticulorumen. Escherichia coli lipopolysaccharide produced intermediates responsible for the smooth muscle relaxation in the first phase of reticuloruminal stasis and for gastric centre depression in the second phase. Adrenergic influences on reticuloruminal motility comprise (a) an alpha1 adrenoreceptor-induced contracture and raised tension receptor sensitivity, (b) an alpha2 adrenoreceptor-mediated depression of the gastric centres causing stasis, excitation of epithelial receptors evoking rumination, and interference with acetylcholine release in the parasympathetic pathway, (c) abeta1 adrenoreceptor-mediated inhibition of the gastric centres, and (d) abeta2 adrenoreceptor-mediated inhibition of intrinsic and extrinsic motility.     

Gastro-intestinal motor disturbances induced by lysine-acetylsalicylate treatment in sheep

The effects of intravenous (i.v.), intramuscular (i.m.) and oral administration of lysine-acetylsalicylate (Lys-ASA) on gastro-intestinal motility were investigated in sheep using electromyography. A dose of 20 mg/kg Lys-ASA intravenously reduced the frequency of reticular contractions for 86 ± 18 min, produced abomasal hypomotility and caused a disruption of the cyclical pattern of intestinal motility for at least 120 min. The frequency of reticular contractions measured from 20 to 30 min after Lys-ASA administration was negatively correlated (ß= 0.97; PΔ0.01) to the log of the dose used for doses varying from 10 to 40 mg/kg. Similar effects were observed with intramuscular and oral dose rates of 40 and 80 mg/kg, respectively. Previous i.v. administration of phentolamine (0.1 mg/kg) or tolazoline (2 mg/kg) abolished the effects of Lys-ASA (20 mg/kg) administered intravenously on both reticular contractions and abomaso-intestinal motility.
It was concluded that Lys-ASA administered at therapeutic doses in sheep produced gastro-intestinal motor disturbances and that α-and α₂-adrenergic antagonists are able to block them.     

The effect of central sympathectomy by 6-hydroxydopamine (6-OHDA) on the response to morphine in conscious sheep

When morphine, an opioid -agonist, was administeredin vivo into the third cerebral ventricle (ICV) of conscious sheep at 20 and 40 µg/kg body weight, it caused psychomotor excitability for 2–3 h and a significant decrease in the reticuloruminal frequency for 45 min and in the mean amplitude of the primary contractions for 65 min. From 60 min after infusion, the same doses of morphine caused a significant increase in the average amplitude of the contractions for 45 min. This suggests that an inhibitory -opioid acceptor is involved in the central control of forestomach motility and general behaviour in sheep. All the effects of morphine were completely prevented by pretreatment with 18.2 µg/kg body weight 6-OHDA ICV. These results suggest that both morphine-induced inhibition of rumen motility and psychomotor excitability are due to central noradrenergic descending system activation. The exact location of the noradrenergic system remains to be determined.     

Depression of reticulo-ruminal motor functions through the stimulation of 012-adrenoceptors

Inhibition of the cyclical contractions of the reticulum and the rumen by detomidine (10-40 micrograms/kg, i.v.), xylazine (20-80 micrograms/kg, i.v.) and clonidine (2.5-10 micrograms/kg, i.v.) were compared in sheep and cattle housed individually in box stalls. Two alpha 2-adrenergic receptor blocking agents, tolazoline and yohimbine, were administered intravenously for prevention (0.1-0.4 mg/kg) or reversal (0.4-1.2 mg/kg) of these effects. Continuous recording of the reticuloruminal contractions and measurement of the volume of ruminal gas eliminated through the upper respiratory tract indicated that the three alpha 2-agonists inhibited the primary ruminal contractions associated with the reticular contractions. The occurrence of secondary ruminal contractions was also blocked in sheep, but only suppressed in cattle. The inhibition of reticulo-ruminal contractions was prevented or reversed competitively by the two alpha 2-blocking agents, suggesting an alpha 2-adrenoceptor mediation of the inhibition of cyclical motor activity of the reticulo-rumen. In contrast with tolazoline, yohimbine was unable to alleviate the accumulation of gas resulting from inhibition of the secondary ruminal contractions.     

Central and peripheral serotonergic control of forestomach motility in sheep

Participation of tryptaminergic receptors in the control of forestomach motility was investigated in conscious sheep using strain-gauges and chronically implanted electrodes. Two hours after feeding the sheep, serotonin (5-HT) was infused into the jugular vein (i.v.), or the carotid artery (i.c.), or into the lateral cerebral ventricles (i.c.v.), over a 10-min period. An i.v. dose of 16 micrograms/kg/min abolished the cyclic propagated contractions throughout the forestomach, increased ruminoreticular tone, and induced simultaneous contractions of all the parts of the rumen. A dose of 1.6 micrograms/kg/min i.c. or i.v. 5-HT inhibited phasic contractions. The effects of 5-HT were blocked completely by i.c.v. administration of methysergide (20 micrograms/kg) and imipramine (200 micrograms/kg), and blocked partially by naloxone (25 micrograms/kg), but unaffected by atropine (50 micrograms/kg). The inhibitory effects of i.v. 5-HT were antagonized by methysergide (200 micrograms/kg, i.v.) but unaffected by imipramine (2 mg/kg, i.v.) and atropine (250 micrograms/kg, i.v.). Only the i.v. administration of methysergide blocked the inhibition induced by i.c. infusion (1.6 micrograms/kg/min) of 5-HT. It is suggested that 5-HT exhibits an inhibitory control on forestomach phasic contractions through hypothalamic and bulbar 5-HT receptors, and exerts peripheral excitatory effects on the tone of the rumen wall.     

Effects of ergotamine and ergovaline on the electromyographic activity of smooth muscle of the reticulum and rumen of sheep

OBJECTIVE: To investigate the effects of IV administration of ergotamine and ergovaline and intraruminal administration of ergotamine on electromyographic (EMG) activity of reticuloruminal smooth muscle in conscious sheep. ANIMALS: 3 sheep with indwelling electrodes in the musculature of the reticulum and rumen. PROCEDURE: In a crossover design study, reticuloruminal motility before and after IV administration of ergotamine (5, 10, 20, and 40 nmol/kg) or ergovaline (2.5, 5, and 10 nmol/kg) was evaluated; EMG effects were compared with those of corresponding control treatments (IV administration of saline [0.9% NaCl] solution or acetone, respectively) in sheep. Ergotamine (800 nmol/kg) or water was also administered intraruminally and their effects compared. RESULTS: After IV administration of ergopeptides, vagally dependent cyclical A and B sequences of contraction of the reticulorumen were immediately inhibited, preceding increases in baseline EMG activity (tonus). The return of cyclical contractions was associated with an increase in contraction amplitude. The effects were dose dependent; administration of 40 nmol of ergotamine/kg resulted in responses that continued for 3 to 4 hours. The effects of intraruminal administration of ergotamine were variable; after 8 hours, EMG activity was increased from baseline for < 2 hours in 1 sheep, 10 hours in another, and > 15 hours in the third. CONCLUSIONS AND CLINICAL RELEVANCE: In sheep, the effects of ergotamine and ergovaline on reticuloruminal motility after IV administration and the duration of responses following intraruminal administration suggest that disruption of digestion may occur in animals grazing endophyte-infected pasture that has a high ergopeptide content.     

Pharmacology of reticulo-ruminal motor function

experimental studies of the pharmacological control of the reticulo-rumen reveal two major central inhibitory pathways: (i) an adrenergic system which inhibits the magnitude of reticulo-ruminal movements and involves alpha 2-receptors and (ii) an inhibitory opioid system, which modulates the rate of the contractions. In addition, the coarseness of the stomach contents and the masticatory movements represent the major excitatory inputs to the gastric centres, hence the hypomotility and stasis observed in disease processes which lead to anorexia. Finally, the amplitude of reticular contractions will tend to vary inversely with the rate of contractions following the administration of stimulatory drugs acting centrally. As a practical statement, a direct action on the gastric centres is possible for drugs such as alpha blockers and/or morphine antagonists. Reflex excitatory effects may be induced by stimulation of muscarinic receptors of the reticulorumen wall as long as it remains below the threshold of a neuromuscular blockade. The highly complex mechanisms whereby reticuloruminal movements are regulated provide for (i) drug interactions, like those of histamine antagonists and opioid agonists, (ii) reflex inhibition of both amplitude and frequency of contractions arising from an enhanced abomaso-duodenal motility, like that induced by serotonin, and (iii) reflex stimulation of the rate of contractions elicited by the i.v. injection of catecholamines, an effect limited to the sheep. The multifactorial nature of reticuloruminal function suggests that treatment is unlikely to effect a cure but should materially re-establish a more normal motility. Further studies, required in diseased animals to provide for a curative use of drugs and a more thorough understanding of drug effects in normal ruminants, represent only a first step toward rational therapy. In addition, the extrapolation of drug effects from sheep to calves or cattle could be hazardous, especially for drugs whose mechanisms of action are currently unknown.     

Vagal reflex inhibition of motility in the abomasal body of sheep by antral and duodenal tension receptors

Vagally-mediated regulation of motility in the abomasal body by duodenal and abomasal antral motility was demonstrated in acutely prepared anaesthetized sheep. The enteric plexuses between the abomasal body, antrum and duodenum were interrupted by transection. Antral contractions were more effective than duodenal contractions at causing inhibition of the abomasal body, and antral isometric conditions were more effective than antral isotonic conditions. Inhibition of motility in the abomasal body was reduced by unilateral cervical vagotomy, was abolished by bilateral cervical vagotomy, and was reversibly inhibited by cervical vagus cold block. The demonstration of vagal pathways in abomaso-abomasal reflexes confirms a functional homology of the mechanisms in the ruminant and animals with simple forms of stomach.Abbreviations b-t balloon-tipped - EMG electromyography - NANC non-adrenergic, non-cholinergic - o-t open-tipped     

Effect of intravenous infusion of proglumide on ruminal motility in conscious sheep (<Emphasis Type="Italic">Ovis aries</Emphasis>)

The effects of intravenous infusion of proglumide on regular ruminal contractions were examined in conscious sheep using doses that inhibit pancreatic exocrine secretion. After a control period of 20 min, proglumide was infused intravenously for 40 min at a dose of 15, 30 or 60 μmol/kg per min and venous blood was collected. The intravenous infusion of proglumide significantly increased the frequency of ruminal contractions at 15 μmol/kg per min without altering the amplitude, while it significantly decreased the frequency and amplitude of ruminal contractions at 30 and 60 μmol/kg per min in a dose-dependent manner. Proglumide did not increase contractile activity of the omasum, abomasum and duodenum or the plasma concentration of immunoreactive cholecystokinin (CCK). Application of proglumide at 1–30 mmol/L inhibited bethanechol-induced contraction in both longitudinal and circular muscle strips of the dorsal sac of the rumen. These results suggest that proglumide at a low dose acts indirectly on the rumen as a CCK receptor antagonist to increase the frequency of contractions, whereas at higher doses it inhibits cholinergic-induced contraction of the ruminal muscles or acts as an agonist to inhibit contractions in sheep. Hence, proglumide at high doses seems unsuitable for research or therapeutic use as a CCK receptor blockade in sheep.     

Beta-endorphin-induced inhibition of rumen contractions in sheep. The effect of hypothalamic de-efferentiation

In conscious sheep, beta-endorphin (1 and 2 micrograms/kg) administered into the third cerebral ventricle caused psychomotor excitability and a significant inhibition of the frequency of rumen contractions. The amplitude of the first rumen contractions, following immediately after the end of endorphin infusion, and the average amplitude of primary rumen contractions were also inhibited. De-efferentiation at the level of the hypothalamus prevented both the inhibitory effect of beta-endorphin on the frequency of rumen contractions and the drug-induced psychomotor excitability. However, de-efferentiation did not prevent beta-endorphin-induced inhibition of the mean amplitude of rumen contractions. The character of pathohistological changes induced by de-efferentiation showed descending degenerative changes of the nerve tracts connecting the hypothalamus with the pons and the medulla oblongata. These results, together with previously published evidence, do suggest that de-efferentiation at the level of the hypothalamus causes degeneration of inhibitory descending opioid-noradrenergic pathways connecting the hypothalamus with the gastric centres in the medulla oblongata.     

Influence of dopamine on rumino-reticular motility and rumination in sheep

The effects of intracerebroventricular (i.c.v.) and intravenous (i.v.) administration of dopamine and its antagonists (domperidone and metoclopramide) on forestomach motility were investigated in four sheep fitted with Nichrome electrodes and strain gauges implanted on the reticulum and the caudo-dorsal sac of the rumen. Infused by the i.c.v. route at a rate of 2 micrograms . kg-1 . min-1, dopamine inhibited the phasic contractions of the reticulo-rumen. A similar effect was obtained following i.v. administration of doses which were 10 times higher, but the effect was associated with an increased tone of the rumen wall. Prior i.v. administration of domperidone (0.5 micrograms . kg-1) blocked these effects and a selective blockade of the dopamine-induced inhibition of phasic contractions was obtained with higher dose of domperidone (2.5 micrograms . kg-1) administered centrally. When dopamine was infused alone ventricularly or intravenously after metoclopramide (40 micrograms . kg-1), it induced a transient (6-8 min) period of rumination, which could be blocked by a prior i.c.v. administration of tolazoline. It was concluded that dopamine acts on rumino-reticular motility in sheep through specific dopamine receptors (i) centrally by inhibiting the frequency of reticular contraction and (ii) peripherally by increasing the muscular tone of the rumen, and its effects on rumination involved alpha 2 adrenergic receptors located in the central nervous system.     

Hexamethonium: A probe to assess autonomic nervous system involvement in upper gastrointestinal functions in conscious sheep

Hexamethonium, which inhibits cholinergic transmission by preventing acetylcholine release, has been considered an ideal reference drug for the blockade of autonomic ganglia, Auerbach plexus and reflex gastrointestinal secretions. The degree of inhibition of ruminant gastrointestinal functions with this reference drug were as follows: cyclical contractions of the reticulo-rumen and abomasal motility > gastric acid secretion and duodenal migrating myoelectrical complexes. Although reduced at high dosages, the initiation of migrating myoelectric complexes was enhanced at clinically used dosages. The duration of the inhibition of reticular contractions was dose-related varying from 0.5 to 5 h for 1.25 to 20 mg/kg subcutaneously. Abomasal motility and acid secretion were similarly reduced but exhibited strong and long-lasting rebound effects. Inhibition of the reticulum by the blockade of muscarinic receptors by atropine was also dose-related lasting from 0.5 to 3 h for 0.5 to 2 mg/kg, whereas inhibition of the abomasal motor and secretory functions lasted from 1 to 6 h. These results suggest a higher degree of impingement of the parasympathetic pathways on abomasal acid secretion and motility than on the cyclical activity of the reticulum and only a modulatory role of the extrinsic neural activity on the cyclical motor events of the duodenum.     

Involvement of α-adrenoreceptors in the regulation of omasal cyclic myoelectrical activity in sheep

In five conscious adult ewes at rest, chronically implanted with electrodes in the musculature of the omasal wall, intravenous (i.v.) infusion for 30 min of alpha 1- or alpha 2-adrenergic receptor blockers, prazosin (20 micrograms/kg/min) and yohimbine (30 micrograms/kg/min), respectively, had no significant effects on omasal myoelectrical activity. The i.v. administration for 15 min of alpha 1- or alpha 2-agonists phenylephrine (4 micrograms/kg/min) or naphazoline (2.5 micrograms/kg/min), respectively, increased the frequency and the amplitude of groups of myoelectrical discharges of omasum, as well as the duration of its activity. Pretreatment of animals with prazosin blocked the responses to phenylephrine. Yohimbine prevented the effects of naphazoline dose-dependently. It is suggested that both alpha 1- and alpha 2-adrenoreceptors are involved in regulation of the sheep's omasal notility. This regulation did not seem to be a simple consequence of the changes in the reticular motility.     

Effects of nitric oxide donor and nitric oxide synthase inhibitor on ruminal contractions in conscious sheep

The present study was planned to evaluate a role of nitric oxide (NO) in the regulation of regular ruminal contractions in conscious sheep. Intravenous infusion of S-nitroso-acetyl-DL-penicillamine (SNAP) at doses of 3-30 nmol kg(-1) min(-1)for 30 minutes inhibited both the amplitude and frequency of ruminal contractions in a dose-dependent manner. However, intravenous infusion of Nomega-nitro-L-arginine-methyl ester (L-NAME) at doses of 0.3-3.0 micromol kg(-1) min(-1)did not alter the basal tone of intraruminal pressure and the amplitude of ruminal contractions. The frequency of contractions was slightly inhibited by L-NAME infusion at 1.0 micromol kg(-1)min(-1). The effects of L-NAME were abolished by simultaneous infusion of L -arginine at 30 micromol kg(-1) min(-1). These results suggest that exogenous NO can diminish the ruminal contractions, while endogenous NO is not involved in the regulatory mechanism of basal tone and regular phasic contractions of the rumen in healthy sheep.     

A method for recording the motor activity of the reticulum in cattle

A method for recording the motility of the reticulum in normal cattle has been devised. The method is based on measurement of the pressure variations occurring in connection with the reticular contractions. The pressure is transferred through open, water-filled catheters via a pressure transducer to an electromanometer, from which it is recorded with the aid of a mingograf.Mean values for the interval, duration and amplitude of the reticular contractions in 10 normal cows are given.The method permits recording in intact animals without any preliminary measures, and can therefore be used in clinical cases.     

Types of serotonergic receptors involved in the control of reticulo-ruminal myoelectric activity in sheep

The effects of peripheral (intravenous, i.v.) and central (intracerebroventricular, ICV) administrations of agonists of 5-HT_1A, 5-HT₂, 5-HT₃ and 5-HT₄ receptors were investigated in conscious sheep chronically fitted with intraparietal electrodes on the reticulum and the dorsal, ventral and caudo-ventral rumen. The 5-HT_1A agonist 8-hydroxydipropylaminotetralin increased reticular and decreased ruminal spike burst frequency when given i.v. (80 μg/kg) and ICV (8 μg/kg). The 5-HT₂ and 5-HT₃ agonists, α-methylserotonin and 2-methylserotonin, induced a moderate inhibition of rumino-reticular contractions when given i.v. at 100 and 150 μg/kg, respectively, while marked inhibition was observed after ICV administration at doses of 10 and 5 μg/kg, respectively. The 5-HT₄ agonist 5-methoxytryptamine strongly stimulated rumino-reticular motility by the ICV (10 μg/kg) route, whereas it induced a moderate inhibition when administered i.v. (200 μg/kg). The selective antagonist of 5-HT_1A, 5-HT₂, 5-HT₃ and 5-HT₄ receptors, spiroxatrine, ritanserin, granisetron and DAU 6285, respectively, blocked the responses of the respective agonists given by the same route. Moreover, the antagonists given ICV blocked the effects of the agonists given i.v. except for DAU 6285 ICV, which did not antagonize the inhibition induced by 5-methoxytryptamine i.v. It is concluded that the four types of serotonergic receptors investigated control rumino-reticular motility at the central level. However, according to the receptor type and the forestomach area (reticulum or rumen) this control may be stimulatory or inhibitory, demonstrating a pleiotropic role of serotonin in the control of rumino-reticular motility in sheep.