Comparison of the inhibitory effects of cromakalim and pinacidil (potassium channel openers) with those of oxybutynin on stimulated guinea pig and rabbit detrusor muscle strips.

To compare the inhibitory effects of a new group of smooth muscle relaxants, the potassium channel openers cromakalim and pinacidil, with those of oxybutynin on detrusor muscle stimulation in animals. Detrusor strips of guinea pigs (n=16) and rabbits (n=20) were mounted in organ bath for recording of isometric tension. Alpha,beta-methylene ATP (10(-7), 10(-6), 10(-5) M), carbachol (10(-6), 10(-5), 3 x 10(-5), 5 x 10(-5) M) and transmural electrical-field stimulation (TES) were applied and concentration-response curves in the absence or presence of cromakalim (10(-6), 10(-5) M), pinacidil (10(-5), 5 x 10(-5) M) and oxybutynin (10(-5), 5 x 10(-5) M) were generated. All curves were displaced to the right in a concentration-dependent manner. The order of potency of inhibition was as follows: alpha,beta-methylene ATP (pinacidil>oxybutynin>cromakalim in guinea pigs; pinacidil>cromakalim>oxybutynin in rabbits); TES (pinacidil>cromakalim>oxybutynin in guinea pigs; cromakalim>oxybutynin>pinacidil in rabbits); carbachol (oxybutynin>pinacidil>cromakalim in guinea pigs; oxybutynin>cromakalim>pinacidil in rabbits). Cromakalim and pinacidil mainly inhibited purinergic-induced (alpha,beta-methylene ATP and TES) detrusor contractions.

Disturbances in the propagation of the slow wave during acute local ischaemia in the feline small intestine.

OBJECTIVE AND DESIGN: The normal aborad propagation of the slow wave in the small intestine is easily distorted by pacing, hypoxia or transection. We studied whether acute local ischaemia would also induce serious conduction disturbances and ectopic pacemaking. METHODS: After general anaesthesia and a mid-abdominal incision, a multi-electrode array of 240 extracellular electrodes was positioned on the serosal surface of an exteriorized intestinal loop. Simultaneous recordings of all 240 surface electrodes was performed during a control period and for 5-10 min following local acute arterial occlusion. After the experiments activation maps were constructed describing the pattern of propagation of the slow waves. RESULTS: During control periods, the activation maps showed homogeneous aborad conduction of the slow wave. During acute ischaemia, local areas of inexcitability developed rapidly, merging together to form lines of conduction block. This in turn often provoked the appearance of subsidiary ectopic pacemakers. The location of the conduction blocks as well as that of ectopic pacemakers was highly variable and could disappear and reappear at other sites. CONCLUSION: Within minutes, acute ischaemia disturbed the organized homogeneous aborad propagation of the slow wave leading to pronounced inhomogeneous depression of conduction, local inexcitability, conduction block and the appearance of subsidiary pacemakers.

Wave mapping: detection of co-existing multiple wavefronts in high-resolution electrical mapping.

High-resolution mapping makes it possible to reconstruct and display the conduction pattern of the action potential as it propagates through cardiac or smooth muscles. During slow and regular activity, time mapping of the spread of activation muscles. During slow and regular activity, time mapping of the spread of activation is relatively simple and straightforward. However, when frequencies are high or conduction is slow, such as seen during atrial fibrillation or found in the pregnant uterus, the tracking of individual waves may become more difficult and uncertain. In order to reconstruct the pathway of a single wave, a search and sorting routine was developed which makes it possible to distinguish, track and display individual wavelets. The algorithm is able to detect variations in conduction block, spontaneous shifts in the location of the pacemaker and changes in the direction of conduction. It is less sensitive when two or more wavefronts intermingle in space and time, such as during collision or fusion. Wave mapping is especially useful, in addition to current time mapping, in sorting quickly through the large amount of data produced by high-resolution mapping of electrical activities in cardiac and smooth muscle.

Spatial and temporal variations in local spike propagation in the myometrium of the 17-day pregnant rat.

Detailed spatial analysis of propagation of individual action potential was performed during spontaneous bursts of activity in the isolated 17-day pregnant rat myometrium. Use was made of high-resolution mapping with simultaneous recordings from 240 extracellular electrodes. Positioning of the electrode assembly by itself did not have any adverse effects, and no differences were found in the period or duration of spontaneous bursts recorded with and without the electrode assembly touching the tissue. The spread of propagation of individual action potentials was reconstructed at several moments during myometrial spike bursts. Both the direction and the sequence of activation of the myometrium were found to be highly variable and depended on 1) the level and spatial dispersion of excitability and 2) whether conduction occurred predominantly in the longitudinal or the circumferential direction. Furthermore, conduction was frequently complicated by the spontaneous occurrence of 1) lines of conduction block, 2) focal sites of pacemaking, or 3) merging of two or more wavelets into a single wave. In contrast, when the myometrium was divided into small segments, activity became much more regular, and both the location of the pacemaker and the direction of propagation were much more stable than in the whole myometrium. In conclusion, spontaneous spatial variations in local spike propagation at the preterm stage could provide for the necessary asynchrony in activation and play a role in the prevention of forceful contractions and premature labor.

Multielectrode mapping of slow-wave activity in the isolated rabbit duodenum.

The technique of multiple simultaneous recordings from a large number of extracellular electrodes (> 100) is currently used in the study of normal and abnormal electrical conduction in the heart and the genesis of cardiac arrhythmias. To investigate whether such a system could also be applied in gastrointestinal electrophysiology, several studies were performed with this technique on segments of isolated rabbit duodenum. A multiple-electrode assembly consisting of 240 silver wires was positioned on the serosal surface of the duodenum, and the recorded signals were, after suitable processing, stored. Thereafter, analysis of all simultaneously recorded slow waves during a selected period of time was performed to reconstruct the pattern of conduction in the duodenum. The first results show that there is a considerable variation in conduction pattern, which is determined by the site of the natural pacemaker. Several experiments were performed to rule out possible deleterious effects of positioning the multiple-electrode assembly on the duodenum. Furthermore, prolonged periods of recording did not influence propagation speed and pattern provided that the positioning of the multiple electrode assembly was performed with care. Entrainment of the natural pacemaker was possible by applying electrical stimuli through 2 of the 240 extracellular electrodes during simultaneous recordings. In conclusion, multisite extracellular mapping of gastrointestinal smooth muscle is possible and can be used to study origin and spread of slow-wave activity.

Hemodynamic and catecholamine responses to laryngoscopy with vs. without endotracheal intubation.

To study the relationship between the intensity of the stimulus exerted against the base of the tongue during direct laryngoscopy and the magnitude of associated hemodynamic and catecholamine responses, a study was conducted in 40 ASA I or II patients. Laryngoscopy lasting 40 s was performed with a size 3 Macintosh blade connected to a force-displacement transducer. The intensity of the stimulus exerted during laryngoscopy is expressed by the product of its average force (N) and duration (s) and given as impulse in Ns. Highly significant relationships were found between the impulse during laryngoscopy and the maximal hemodynamic and catecholamine responses. Also, when laryngoscopy was followed by orotracheal intubation, significant relationships were found with steeper slopes of the regression lines for systolic blood pressure, heart rate and plasma epinephrine concentrations. A more rapid regression of hemodynamic data was seen in intubated patients, whereas their plasma catecholamine concentrations regressed more slowly. The mechanisms of the responses to laryngoscopy and orotracheal intubation are proposed to be by somato-visceral reflexes. Stimulation of proprioceptors at the base of the tongue during laryngoscopy induces impulse-dependent increases of systemic blood pressure, heart rate and plasma catecholamine concentrations. Subsequent orotracheal intubation recruits additional receptors that elicit augmented hemodynamic and epinephrine responses as well as some vagal inhibition of the heart.

Vascular effects of some opioid receptor agonists.

Opioid peptides have been implicated in shock-associated hypotension. Our aim was to find out whether opioid agonists have direct vasodilator actions on vascular smooth muscle. The study was conducted on rat abdominal aortic rings. In rings precontracted with either norepinephrine, prostaglandin F2 alpha, or high potassium Krebs (HPK), the effects of the opioid agonists tested (morphine, U50488H, ethylketocyclazocine (EKC), and bremazocine) depended on the precontracting agent used. HPK-precontracted rings were relaxed by all agonists tested. In norepinephrine-precontracted rings, all caused contraction at low concentrations and relaxation at high concentrations except bremazocine, which caused only relaxation. In prostaglandin F2 alpha-precontracted rings, U50488H produced contraction at low concentrations and relaxation at high concentrations while EKC caused only relaxation and morphine or bremazocine caused only contraction. All relaxant responses were endothelium-independent and were antagonized by verapamil but not by a number of antagonists including naloxone. MR2266, propranolol, diphenhydramine, cimetidine, and indomethacin. They may reflect calcium channel blockade. Morphine-induced vasoconstriction was antagonized by high concentrations of of naloxone or mepyramine and may be due to release of histamine by a naloxone-sensitive mechanism. We conclude that (a) the opioid agonists tested exert direct actions on vascular smooth muscle; (b) the nature of the response depended not only on the agonist used and its concentration but also on the agent used to precontract the tissue; and (c) it is unlikely that direct actions of endogenous opioids contribute to the shock-associated hypotension because high doses were needed to elicit them.

Electrical basis of excitation and inhibition of human colonic smooth muscle.

Excitation and inhibition of electrical activities of the musculature of the human colon and the consequent changes in motor activities were studied in vitro. The mechanisms of excitation and inhibition were very different from those of the small intestine and colons from animal models. Carbachol increased spiking activity and the frequency of bursts of electrical oscillations in longitudinal muscle. Each longitudinal muscle contraction was related to a burst of electrical oscillations. Carbachol induced one of three patterns of activity in circular muscle: (a) continuous electrical oscillatory activity (14-24 cpm) with spikes, associated with tonic contraction; (b) bursts of such electrical activity, associated with broad phasic contractions; or (c) prolonged membrane potential depolarizations (frequency 1-3 cpm) with superimposed intense spiking activity, associated with phasic contractions. Isoproterenol inhibited all electrical and mechanical activities in both muscle layers. These results may provide a better understanding of (a) the origins of the variable patterns of electrical and motor activities and (b) the relationship between electrical and mechanical activities of the human colon musculature.

Vagal control of canine postprandial upper gastrointestinal motility.

The role of the vagus nerves in the control of postprandial motility in the upper gastrointestinal tract was investigated in four dogs by use of a bilateral cervical cooling blockade technique. On administration of food, the fasting migrating motor complex (MMC) was replaced by the postprandial (feeding) pattern. Feeding pattern duration varied in a dose-dependent manner with either total volume or calories of food. During the feeding pattern, oscillations in lower esophageal sphincter (LES) pressure occurred at time intervals equivalent to the MMC cycle period. Twenty-one control feeding experiments and 17 postprandial vagal blockade experiments were performed, with a minimum of three of each type in each dog. Vagal blockade, initiated at times ranging from 15 min to 4 h after feeding and maintained for up to 5 h, abolished the postprandial activity in the upper gastrointestinal tract. During postprandial vagal blockade, LES pressure was abolished and bursts of contractions were observed only in the upper small bowel, a pattern resembling that observed during vagal blockade in the fasted state. These bursts occurred at the expected times relative to, and their cycle period was not significantly different from, that of the MMCs recorded prior to feeding. Vagal blockade started prior to feeding prevented initiation of the fed pattern, which appeared immediately on termination of the blockade. We conclude that initiation and maintenance of the postprandial pattern in the upper gastrointestinal tract with concurrent inhibition of the fasting MMC normally require vagal integrity. The "clock" controlling the MMC cycle period is not reset by feeding, but its effect on motility is suppressed.

The Ehlers-Danlos syndrome and colonic perforation. Report of a case and physiologic assessment of underlying motility disorder.

The Ehlers-Danlos syndrome is a genetically determined disorder of connective tissue which is generally known for its features of fragile, hyperextensible skin, hypermobile joints, and tissue fragility. Less commonly, colorectal complications can occur, including bleeding, prolapse, and diverticulitis. A rare case of colonic perforation associated with Ehlers-Danlos syndrome is presented. Additionally, in vitro electromyographic studies of the colonic tissue were performed which suggested a possible link between abnormal myogenic activity and the colonic perforations. The authors recommend that treatment be either a permanent colostomy or a subtotal colectomy with anastomosis to the rectum for similar cases.

Electrophysiologic control of motility in the human colon.

Characteristics of electrical activities, and the relationship between electrical and motor activities, were studied in circular and longitudinal (taenia) muscle of the human colon that was obtained from 21 individuals. Recordings were obtained with suction electrodes, the sucrose-gap method, and microelectrodes. The circular muscle electrical activity consisted of oscillatory activity of relatively low amplitude, with a frequency range from 4.5 to 60 cycle/min. Spiking activity was present on most oscillations. Contractile activity was associated with individual oscillations at frequencies below 12 cycle/min. Contractions related to periods of oscillations at frequencies above 12 cycle/min showed summation resulting in prolonged contractions. In these periods, oscillations were either of relatively high amplitude, or had superimposed spiking activity. Longitudinal muscle activity consisted of slow electrical oscillations at frequencies between 24 and 36 cycle/min with spiking activity superimposed on most oscillations. Contractions were related to bursts of such activity. These findings provide the electrophysiologic basis for short and prolonged phasic contractions and for sustained contractions of the human colon muscle layers. Activities in both muscle layers were myogenic in nature, were very sensitive to stretch, and could be initiated or modulated by nervous activity.

Oesophageal peristalsis in the cat: the role of central innervation assessed by transient vagal blockade.

Studies were performed on five cats to assess the role of extrinsic vagal innervation in the control of peristalsis in the smooth muscle oesophagus. Transient vagal nerve blockade was accomplished by cooling the cervical vagosympathetic nerve trunks previously isolated in skin loops on each side of the neck. Peristalsis throughout the body of the oesophagus was monitored using a continuously perfused multilumen manometry tube. Striated and smooth muscle portions of the esophagus were delineated by abolishing smooth muscle activity with atropine. Secondary peristalsis was assessed by intra-oesophageal balloon distension studies. The threshold volume for balloon-induced secondary peristalsis was lower in the smooth muscle oesophagus. Unilateral vagal blockade reduced the incidence of primary and secondary peristalsis in the striated muscle oesophagus but not in the smooth muscle oesophagus. Bilateral vagal nerve blockade abolished primary swallow-induced peristalsis and secondary peristalsis in both the smooth and striated muscle cat oesophagus. Administration of cholinergic agents or adrenergic blocking agents failed to restore secondary peristalsis in the smooth muscle oesophagus during vagal cooling. We conclude that connections to the central nervous system via the vagal nerve trunks are required for normal secondary as well as primary peristalsis in both the smooth and striated muscle portions of the cat oesophagus.

Electrophysiological basis of excitation of canine colonic circular muscle by cholinergic agents and substance P.

The circular muscle layer of the canine colon exhibits omnipresent rhythmic periodic waves of depolarization (slow waves), acting as pacemaker activity. The electrophysiological and motor responses of this layer to the muscarinic agonists acetylcholine and carbachol, and to the excitatory peptide substance P, were studied using the sucrose-gap technique. In addition, changes in the contractile activity were examined in organ bath experiments. The slow waves consisted of an initial potential followed by a plateau potential. All substances depolarized the membrane and increased dramatically the duration of the plateau potential resulting in a decrease of the slow wave frequency. In addition, the amplitude of the plateau potential was often increased significantly. Carbachol and substance P readily evoked spiking activity whereas acetylcholine did not. Both spiking activity and the plateau potential generated contractile activity. The prolongation of the slow wave duration caused a profound alteration of the pattern of contractions. Long-lasting tachyphylaxis to the effect of substance P, but not to acetylcholine or carbachol, occurred. The electrophysiological and motor effects of the drugs were due to a direct action on the smooth muscle cell membrane. This study provides an electrophysiological basis for prolonged circular muscle contractions of the colon, and it emphasizes the pacemaker activity of gastrointestinal smooth muscle as an important site of drug action.

The effects of cholecystokinin-octapeptide and pentagastrin on electrical and motor activities of canine colonic circular muscle.

The effects of cholecystokinin-octapeptide (CCK-OP) and pentagastrin on electrical and motor activities of circular muscle of the canine colon were studied with the sucrose gap technique. Additional organ bath experiments were performed to further characterize the motor response to the peptides and to elucidate their site of action. The electrical activity consisted of slow waves having an initial potential followed by a plateau potential, at a regular frequency of 4.5 cycles/min. Both peptides prolonged the duration and increased the amplitude of the plateau phase of the slow waves. Concomitantly, the slow wave frequency was reduced. In addition, CCK-OP increased spiking activity. Both spiking activity and the prolonged plateau potential generated contractile activity, prolonged phasic contraction occurring with slow waves with a prolonged plateau. In organ bath experiments, both CCK-OP and pentagastrin increased the basal tone of the muscle strips and prolonged the duration of the phasic contractions. The prolongation of the duration of the contractions was not antagonized by tetrodotoxin (TTX) and atropine. CCK-OP but not pentagastrin increased the force of contractions, this action was not affected by atropine but was reduced in the presence of TTX, suggesting that the increase in force may be partially mediated by noncholinergic excitatory nerves. The increase in basal tension by the peptides was enhanced in the presence of TTX indicating that myenteric inhibitory neurones were tonically active under our experimental conditions. The results provide the electrophysiological basis for CCK-OP and pentagastrin induced changes in colonic motility.

Fasting canine biliary secretion and the sphincter of Oddi.

This study correlates duodenal bile acid delivery with motility of the sphincter of Oddi during the fasting state. Dogs were prepared with a functional cholecystectomy, a duodenal cannula for direct vision cannulation of the common bile duct, and 12 bipolar electrodes serosally implanted from stomach to terminal ileum. In one set of experiments, the bile acid pool was depleted, and during a continuous i.v. infusion of sodium taurocholate (20 mumol/min), duodenal bile acid delivery was assessed over 6 h by a marker perfusion technique. In other experiments, a double-lumen continuously perfused manometry catheter was placed to record motility in the bile duct and sphincter of Oddi for a period of 6 h. Station pull-throughs of the sphincter of Oddi were performed in each phase of the migrating motor complex. Bile acid secretion rates fluctuated about the i.v. infusion rate during duodenal phase I and II, peaked in late phase II, and then fell to barely detectable levels during duodenal phase III. There was no peristaltic contractile activity in the common bile duct in any phase of the migrating motor complex. The sphincter of Oddi maintained a baseline pressure above common bile duct pressure. It was highest during phase III. Phasic contractions of the sphincter of Oddi were intermittent during phase I, increasingly frequent during phase II, and continuous during duodenal phase III of the migrating motor complex. Contractions were frequently peristaltic. We concluded that the occurrence and amplitude of phasic peristaltic contractions of the sphincter of Oddi are cyclically coordinated with the fasting intestinal motor pattern (migrating motor complex), and with cyclical variations in the delivery of bile acids into the duodenum. Both resting pressure and phasic contractions of the sphincter appear to play a role in coordinating the cyclic delivery of bile acids into the duodenum with the migrating motor complex. Intense phasic motor activity appears to impede bile flow, and less intense activity allows or facilitates flow.

Relationship between porcine motilin-induced migrating motor complex-like activity, vagal integrity, and endogenous motilin release in dogs.

Both the vagus nerve and motilin have been implicated in the initiation of phase III of the fasting migrating motor complex. To investigate the contribution of each, the effects of intravenous porcine motilin, vagosympathetic nerve blockade, and atropine were assessed. Intralumenal pressures of the lower esophageal sphincter, stomach, and upper small intestine, and plasma motilin levels were monitored. Porcine and canine motilin in plasma were distinguished by radioimmunoassay using two different antibodies. Injection of porcine motilin (75-370 pmol), initiated dose-dependent phase III-like motor activity in the lower esophageal sphincter, stomach, and small bowel if the vagi were intact; if the vagi were blocked, activity was initiated in the small bowel only. Moreover , a significant (p less than 0.001), dose-dependent peak in canine plasma motilin was observed after the onset of the induced motor activity when the vagi were intact or blocked, with plasma motilin peaks comparable to those occurring spontaneously. In both intact and vagally blocked dogs, atropine abolished both the spontaneous motor activity and associated rise in motilin level, and also abolished porcine motilin-induced activity. However, a diminished, but significant (p less than 0.01) peak in porcine motilin-induced canine motilin persisted in the presence of atropine. These results in dogs indicate that while spontaneous phase II motility in the upper gastrointestinal tract and phase III activity in the lower esophageal sphincter and stomach are dependent on vagally mediated excitatory pathways, spontaneous and induced phase III motor activity in the small intestine are dependent on nonvagal cholinergic innervation. Canine motilin release induced by porcine motilin is mediated primarily by a nonvagal cholinergic (muscarinic) pathway, with minor contributions from vagal noncholinergic, and nonvagal noncholinergic mechanisms. Because spontaneous or induced motilin release peaks well after the onset of phase III motor activity, it is unlikely that motilin is the primary factor responsible for initiation of the migrating motor complex in the dog. Motilin may, however, modulate motility produced by preexisting neural excitation.

Lower esophageal sphincter function in the cat: role of central innervation assessed by transient vagal blockade.

Studies were performed on four cats to assess the role of extrinsic vagal innervation in the control of lower esophageal sphincter (LES) function. Both cervical vagal nerves were blocked transiently by cooling. LES pressure was measured using a multilumen manometry tube. LES relaxation was assessed during intraesophageal balloon distension in both the striated and smooth muscle portions of the esophagus. Bilateral vagal nerve blockade lowered the mean LES pressure from 58 +/- 17 to 29 +/- 9 mmHg (P less than 0.01). During vagal blockade, balloon distension in the striated muscle esophagus further reduced sphincter pressure to 16 +/- 4 mmHg (P less than 0.01) and that in the smooth muscle esophagus to 15 +/- 3 mmHg (P less than 0.01). Swallow-induced LES relaxation was abolished during bilateral vagal nerve blockade. During vagal blockade, atropine reduced LES pressure to 10 +/- 1 mmHg, phentolamine to 13 +/- 6 mmHg, and hexamethonium to 10 +/- 4 mmHg (all P less than 0.01). We conclude that 1) normal LES tone in the cat is mediated primarily by two separate neural mechanisms: a vagal cholinergic mechanism and a nonvagal mechanism that utilizes both alpha-adrenergic and cholinergic receptors; 2) local, intramural mechanisms of high threshold are present in the striated and smooth muscle cat esophagus to allow distension-induced reflex inhibition of the LES; and 3) swallow-induced LES relaxation is dependent on vagally mediated central nervous system connections.

Interaction of coupled nonlinear oscillators having different intrinsic resting levels.

The interaction among coupled oscillators is governed by oscillator properties (intrinsic frequency and amplitude) and coupling mechanisms. This study considers another oscillator property, the intrinsic resting level, and evaluates its role in governing oscillator interactions. The results of computer experiments on a chain of either three or five bidirectionally coupled nonlinear oscillators, suggest that an intrinsic resting level gradient, if present, is one of the factors governing the interaction between coupled oscillators. If there is no intrinsic frequency gradient, then an intrinsic resting level gradient is sufficient to produce many features of interaction among coupled oscillators. If both intrinsic frequency and intrinsic resting level gradients are present, then both of them determine the manner in which the coupled oscillators interact with each other.

Vagal control of migrating motor complex-related peaks in canine plasma motilin, pancreatic polypeptide, and gastrin.

The role of the vagus nerve in the control of fasting plasma pancreatic polypeptide (PP), gastrin, and motilin levels was investigated in conscious dogs. Lowest plasma levels of motilin (81 +/- 8 pmol/L), PP (19 +/- 1 pmol/L) and gastrin (5 +/- 1 pmol/L) were observed during phase I of the migrating motor complex (MMC). Significant peaks in plasma motilin (127 +/- 11 pmol/L, P less than 0.005), PP (26 +/- 2 pmol/L, P less than 0.005), and gastrin (14 +/- 2 pmol/L, P less than 0.005) were seen, coinciding with the appearance of phase II (PP and gastrin) or phase III (motilin) of the migrating motor complex in the upper gut. Whereas bilateral vagal blockade abolished the peaks in PP and gastrin, a significant (P less than 0.025) increment in plasma motilin remained, which correlated with the late phase III equivalent of the vagally independent complex (VIC) in the duodenum. This VIC-related motilin peak (170 +/- 20 pmol/L) was significantly higher (P less than 0.025) and the time course (9 +/- 2 min) significantly shorter (P less than 0.01) than the peak (127 +/- 11 pmol/L) and duration (31 +/- 9 min) observed without vagal blockade. Thus, in fasting, the cyclical increments of PP and gastrin are both dependent on excitatory vagal innervation, whereas excitatory pathways controlling phase III associated peak motilin release are nonvagal. In addition, the pattern of fasting motilin release and the amplitude of peak motilin secretion may be affected by vagal inhibition.