Spatiotemporal mapping of the motility of the isolated chicken caecum

We studied the caecal contractile activity of the chicken (Gallus gallus) using single caeca that had been cannulated at their proximal and distal ends, and in paired caeca, maintained in situ on excised segments of gut that were cannulated at the colonic and small intestinal ends. Longitudinal and circular contractile patterns were characterised using high-definition spatiotemporal mapping. Low amplitude longitudinal contraction waves of frequency 14.1 cycles/min occurred in the absence of major contractile events. These were termed fast phasic and appeared to be mediated by slow waves. The nature of major spontaneous contractions occurring in the single caecum varied with the level of caecal distension. Type A contractions occurred when the caecum was not distended, originated from variable sites and propagated in both directions. Type B or C contractile events occurred when the caecum was moderately or fully distended, originated from a predominantly distal site and propagated proximally. On diameter maps, each type B event comprised a succession of contractions which had similar propagation speeds, frequency and direction to fast phasic contractions. Type C events were comprised of a succession of higher amplitude contractions with no appreciable propagation. Perfusion of saline via the colon resulted in fluid entering both caeca and the onset of aborad contractions in their proximal canals. Saline was also seen to flow between caeca during contractile events however no saline was seen to enter the small intestine as has been postulated by other workers.     

High definition mapping of circular and longitudinal motility in the terminal ileum of the brushtail possum <Emphasis Type="Italic">Trichosurus vulpecula</Emphasis> with watery and viscous perfusates

Longitudinal and radial movements during spontaneous contractions of isolated segments of terminal ileum of the brushtail possum, a species of arboreal folivore, were studied using high definition spatiotemporal maps. Segments obtained from specimens were continuously perfused with solutions of various apparent viscosities at 3 cm and 5 cm hydrostatic pressure. A series of sustained tetrodotoxin-sensitive peristaltic events occurred during perfusion. The leading edge of each peristaltic event progressed by a succession of rhythmic surges of circular contraction with concerted concurrent phasic longitudinal contractions. Three types of peristaltic event were observed, with differing durations of occlusion and patterns of cyclic, in phase, circular and longitudinal contractions. Each peristaltic event was preceded by a change of shade on the D map that indicated circumferential dilatation. Differences in the slopes of these phasic shade changes from those occurring during peristalsis indicate that this distension is passive and likely results from aboral displacement of fluid. Tetradotoxin insensitive longitudinal contraction waves of frequency 9.2 min⁻¹ occurred during and in the absence of peristalsis, originating at a variety of sites, and propagating either in an orad or aborad direction but predominantly in the latter. Perfusion with 1% guar gum, at 5 cm hydrostatic pressure caused the lumen to become distended and the generation of peristaltic events to cease pending reduction of the hydrostatic head to 3 cm but longitudinal contractile activity was preserved. Neither the frequencies nor the rates of progression of circular and longitudinal contractile events, nor the temporal coordination between these events, varied with the apparent viscosity of the perfusate or altered in a manner that could facilitate mixing.     

High-definition spatiotemporal mapping of contractile activity in the isolated proximal colon of the rabbit

Four types of contractile activity were identified and characterised in the isolated triple haustrated proximal colon of the rabbit using high-definition spatiotemporal mapping techniques. Mass peristalses were hexamethonium-sensitive deep circular contractions with associated taenial longitudinal contractile activity that occurred irregularly and propagated rapidly aborad, preceded by a zone of local lumen distension. They were sufficiently sustained for each event to occupy the length of the isolated colonic segment and the contraction persisted longer orally than aborally, the difference being more pronounced when lumen contents were viscous. Haustra were bounded by deep even-spaced ring contractions that progressed slowly aborad (haustral progression). Haustral formation and progression were hexamethonium-sensitive and coordinated across intertaenial domains. Ripples were hexamethonium-resistant phasic circular contractions that propagated predominantly orad at varying rates. In the presence of haustra, they were uncoordinated across intertaenial domains but were more coordinated when haustra were absent. Fast phasic contractions were relatively shallow hexamethonium-resistant contractions that propagated rapidly in a predominantly aborad direction. Fast phasic circular contractions were accompanied by taenial longitudinal muscle contractions which increased in amplitude prior to a mass peristaltic event and following the administration of hexamethonium. On the basis of the concurrence and interaction of these contractile activities, we hypothesise that dual pacemakers are present with fast phasic contractions being modulated by the interstitial cells of Cajal in the Auerbach’s plexus (ICC-MY) while ripples are due to the submucosal ICC (ICC-SM). Further, that ICC-SM mediate the enteric motor neurons that generate haustral progression, while the intramuscular ICC (ICC-IM) mediate mass peristalsis. The orad movement of watery fluid was possibly due to ripples in the absence of haustra.     

The mechanical advantage of local longitudinal shortening on peristaltic transport

Whereas bolus transport along the esophagus results from peristaltic contractions of the circular muscle layer, it has been suggested that local shortening of the longitudinal muscle layer concentrates circular muscle fibers in the region where the highest contractile pressures are required. Here we analyze the mechanical consequences of local longitudinal shortening (LLS) through a mathematical model based on lubrication theory. We find that local pressure and shear stress in the contraction zone are greatly reduced by the existence of LLS. In consequence, peak contractile pressure is reduced by nearly 2/3 at physiological LLS, and this reduction is greatest when peak in LLS is well aligned with peak contractile pressure. We conclude that a peristaltic wave of local longitudinal muscle contraction coordinated with the circular muscle contraction wave has both a great physiological advantage (concentrating circular muscle fibers), and a great mechanical advantage (reducing the level of contractile force required to transport the bolus), which combine to greatly reduce circular muscle tone during esophageal peristalsis.     

Synchrony between circular and longitudinal muscle contractions during peristalsis in normal subjects

The current understanding is that longitudinal muscle contraction begins before and outlasts circular muscle contraction during esophageal peristalsis in normal subjects. The goal of our study was to reassess the relationship between the contractility of two muscle layers using novel ways to look at the muscle contraction. We studied normal subjects using synchronized high-frequency ultrasound imaging and manometry. Swallow-induced peristalsis was recorded at 5 and 10 cm above the lower esophageal sphincter (LES). Ultrasound (US) images were analyzed for muscle cross-sectional area (CSA) and circularity index of the esophagus during various phases of esophageal contraction. A plot of the M mode US image, muscle CSA, and esophageal circularity index was developed to assess the temporal correlation between various parameters. The muscle CSA wave began before and lasted longer than the contraction pressure wave at both 5 and 10 cm above the LES. M mode US images revealed that the onset of muscle CSA wave was temporally aligned with the onset of lumen collapse. The peak muscle CSA occurred in close proximity with the peak pressure wave. The esophagus started to become more circular (decrease in circularity index) with the onset of the muscle CSA wave. The circularity index and muscle CSA returned to the baseline at approximately the same time. In conclusion, the onset of lumen collapse and return of circularity index of the esophagus are likely to be the true markers of the onset and end of circular muscle contraction. Circular and longitudinal muscle layers of the esophagus contract in a precise synchronous fashion during peristalsis in normal subjects.     

江豚(Neophocaena asiaeorientalis)的消化器官Ⅱ. 肠、肝、胰

本篇为江豚(Neophocaena asiaeorientalis)消化器官研究的第二部分。 标本来源见李悦民(1984)。肠的长度和肝的重量都由新鲜标本测得。组织学材料用10%甲醛溶液固定,石蜡包埋,切片用苏木精曙红,PAS和Van Gieson法染色。     

Local longitudinal muscle shortening of the human esophagus from high-frequency ultrasonography

We analyzed local longitudinal shortening by combining concurrent ultrasonography and manometry with basic principles of mechanics. We applied the law of mass conservation to quantify local axial shortening of the esophageal wall from ultrasonically measured cross-sectional area concurrently with measured intraluminal pressure, from which correlations between local contraction of longitudinal and circular muscle are inferred. Two clear phases of local longitudinal shortening were observed during bolus transport. During luminal filling by bolus fluid, the muscle layer distends and the muscle thickness decreases in the absence of circular or longitudinal muscle contraction. This is followed by local contraction, first in longitudinal muscle, then in circular muscle. Maximal longitudinal shortening occurs nearly coincidently with peak intraluminal pressure. Longitudinal muscle contraction begins before and ends after circular muscle contraction. Larger longitudinal shortening is correlated with higher pressure amplitude, suggesting that circumferential contractile forces are enhanced by longitudinal muscle shortening. We conclude that a peristaltic wave of longitudinal muscle contraction envelops the wave of circular muscle contraction as it passes through the middle esophagus, with peak longitudinal contraction aligned with peak circular muscular contraction. Our results suggest that the coordination of the two waves may be a physiological response to the mechanical influence of longitudinal shortening, which increases contractile force while reducing average muscle fiber tension by increasing circular muscle fiber density locally near the bolus tail.     

Effects of GABA on the spontaneous muscular activity of the human fallopian tube ampullar segments in vitro

The effects of gamma-aminobutyric acid (GABA) and some GABA ergic compounds were examined on the spontaneous contractile activity of the ampullar segment of human Fallopian tubes in vitro. In both circular and longitudinal smooth muscle layers, GABA increased the frequency, and decreased the amplitude of the contractions. In longitudinal muscle layer, this effect occurred independently of endocrine status, while in the circular muscle layer, the amplitude was affected only in follicular phase and in postmenopause, but not in the luteal phase. Baclofen mimicked the effects of GABA, whereas muscimol and bicuculline failed to influence the contractility. Our findings suggest that, in the ampulla of the human oviduct, GABAB-receptors may be involved in the regulation of motility, and thus in the modulation of ovum transport and fertility.     

Pressure morphology of the relaxed lower esophageal sphincter: the formation and collapse of the phrenic ampulla

This study aimed to apply novel high-resolution manometry with eight-sector radial pressure resolution (3D-HRM technology) to resolve the deglutitive pressure morphology at the esophagogastric junction (EGJ) before, during, and after bolus transit. A hybrid HRM assembly, including a 9-cm-long 3D-HRM array, was used to record EGJ pressure morphology in 15 normal subjects. Concurrent videofluoroscopy was used to relate bolus movement to pressure morphology and EGJ anatomy, aided by an endoclip marking the squamocolumnar junction (SCJ). The contractile deceleration point (CDP) marked the time at which luminal clearance slowed to 1.1 cm/s and the location (4 cm proximal to the elevated SCJ) at which peristalsis terminated. The phrenic ampulla spanned from the CDP to the SCJ. The subsequent radial and axial collapse of the ampulla coincided with the reconstitution of the effaced and elongated lower esophageal sphincter (LES). Following ampullary emptying, the stretched LES (maximum length 4.0 cm) progressively collapsed to its baseline length of 1.9 cm (P < 0.001). The phrenic ampulla is a transient structure comprised of the stretched, effaced, and axially displaced LES that serves as a "yield zone" to facilitate bolus transfer to the stomach. During ampullary emptying, the LES circular muscle contracts, and longitudinal muscle shortens while that of the adjacent esophagus reelongates. The likely LES elongation with the formation of the ampulla and shortening to its native length after ampullary emptying suggest that reduction in the resting tone of the longitudinal muscle within the LES segment is a previously unrecognized component of LES relaxation.     

Neuromuscular control systems in the proboscis of Paranemertes peregrina

Neural systems underlying muscular control in the proboscis of Paranemertes peregrina were investigated through the effects of various neuropharmacological agents on electrical and contractile activity.

Peristalsis in the circular muscle was elicited by catecholaminergic drugs. Each wave was accompanied by a spindle‐shaped burst of electrical activity. The occurrence and rate of peristalsis could be manipulated separately, suggesting the presence of a polyneuronal pathway. Acetylcholine elicited contraction of the longitudinal musculature, with little or no obvious accompanying electrical activity. The circular and longitudinal control systems exhibited reciprocal inhibition, which appeared to occur prior to the postsynaptic neuromuscular junction. Additionally, both systems were excited by serotonin.

Histochemical localisations revealed cholinesterases in the longitudinal musculature. Norepinephrine in the proboscideal nerve cords and serotin in the proboscideal nerve cords and plexus were demonstrated through fluorescence spectrophotometry.     

Evaluation of peristalsis in multiple segments of the guinea-pig isolated small intestine: optimisation of tissue use by refined in vitro methodology

Peristalsis is the aboral movement by which the intestine propels its contents. Since pharmacological research requires an experimental model with which drug-induced modifications of peristalsis can be reliably quantified, we set out to develop and validate an in vitro method for studying peristalsis in multiple gut segments. In our arrangement, up to four 10cm segments isolated from the guinea-pig jejunum and ileum can be set up in parallel and their lumens perfused. Peristalsis was elicited by pressure-evoked wall distension, and the peristalsis-induced changes in the intraluminal pressure were evaluated with software that determined the peristaltic pressure threshold, the frequency, maximal acceleration and amplitude of the peristaltic waves, and the residual baseline pressure. Validation experiments showed that the peristalsis parameters at baseline and after modification by morphine (0.01-10microM) did not differ between segments from the jejunum and ileum, or between segments examined in a consecutive manner. In conclusion, our work succeeded in optimising the use of the guinea-pig jejunum and ileum for multiple recordings of peristalsis in vitro, and in refining the recording and evaluation of peristaltic motility. This system promises to be particularly useful in the pharmacological screening and testing of drugs which modify peristalsis.     

Stress-induced inhibition of the ileal, caecal, and colonic contractile activity in rabbits]

In alert rabbits, immobilisation stress decreased the spike burst rate and amplitude in ileum, caecum, and colon for 20 min. Following beta-adrenoceptor blockade, the contractile activity suppression was aggravated. The stress seems to lead to suppression of the ileal, caecal, and colonic contractile activity for up to 40 min which is unrelated to adrenergic inhibition of the smooth muscle activity mediated by beta-adrenoceptors.     

Tachykinins mediate non-adrenergic,non-cholinergic excitatory neurotransmission to the hamster ileum via NK1 and NK2 receptors

The present study was designed to investigate Substance P (SP) and a related tachykinin, Neurokinin A (NKA), contributions to the excitatory neurotransmission to the circular smooth muscle of the hamster ileum. In the presence of atropine (0.5 microM), guanethidine (3 microM) and NG-nitro-L-arginine methyl ester (L-NAME) (200 microM), electrical field stimulation (EFS) evoked a non-adrenergic, non-cholinergic (NANC) excitatory junction potential (EJP) and contraction of circular smooth muscle. Applications of SP and NKA produced depolarizing and contractile responses in a concentration-dependent fashion. The EJP and contraction were almost abolished by the non-specific tachykininergic antagonist, spantide (3 microM). Application of SP antagonist, L-732,138, (1 microM) markedly inhibited EJP (82.5%) and contraction (68.9%) and completely blocked excitatory responses produced by exogenous application of SP. While application of NKA antagonist, SR48968 (1 microM) completely blocked the depolarising and contractile responses to NKA, it only slightly inhibited those to EFS (17.2% and 31.4% respectively).These results provide evidence that, in the circular muscle of hamster ileum, endogenous tachykinins are the main NANC excitatory neurotransmitters and their action is mediated by both NK1 and NK2 receptors.     

Comparative effects of galanin on isolated smooth muscle cells from ileum in five mammalian species.

Effect of galanin and CCK8 were studied on isolated smooth muscle cells obtained from pig, guinea-pig, rat, rabbit and dog ileum circular muscle layer. Galanin as well as CCK8 induced a concentration-dependent contraction of pig, rat, rabbit and guinea-pig ileum smooth muscle cells. Maximal contraction ranged between 23.7 +/- 1.9% and 26.1 +/- 3.1% decrease in cell length from control in the presence of both peptides. This maximal contraction was obtained at 1 nM galanin in pig, rat, rabbit, 1 nM CCK8 in rat, rabbit, guinea-pig, at 10 nM galanin in guinea-pig and 10 nM CCK8 in pig. Concentrations of galanin inducing a half maximal contraction (EC50) ranged between 8 pM and 80 pM in these species. In dog, CCK8 induced a concentration-dependent contraction of ileum smooth muscle cells, with a maximal contraction (24.5 +/- 2.3%) at 1nM and an EC50 of 50 pM while galanin inhibited cell contraction induced by CCK8. The CCK-induced contraction was abolished at 10 nM galanin and 10 nM VIP. Concentrations of galanin and VIP inducing a half-maximal relaxation of contracted cells were 2 pM and 3 pM respectively. It is concluded that galanin may induce cell contraction of pig, guinea-pig, rat and rabbit ileum circular muscle layer and cell relaxation of dog ileum by a direct myogenic effect.     

Endogenous opioid peptide effects on the guinea-pig biliary tract

The effects of the endogenous opioid peptides, dynorphin (1-13), Met-enkephalin and beta-endorphin, on contraction induced by transmural stimulation of terminal bile duct preparations (terminal cavity and ampulla) and gallbladder were investigated in vitro. These peptides inhibited ampulla contraction, dose dependently. The potency order, indicated by ID50, was the same as in the guinea-pig ileum, but the absolute ID50 values in the ampulla were lower than in the ileum. In the terminal cavity, the dynorphin (1-13) ID50 was still less than in the ampulla, and beta-endorphin and Met-enkephalin did not reduce contraction by as much as 50%. In the gallbladder, the effects of these opioid peptides on contraction induced by transmural stimulation were not significant. The results suggest differences in the receptor populations of the ampulla and terminal cavity, and lack of opiate receptors (at least micro and k receptors) in the gallbladder.     

Endogenous prostaglandins regulate spontaneous contractile activity of uterine strips isolated from non-pregnant pigs

Myometrial strips isolated from non-pregnant pigs show spontaneous contractile activity. In the present study, the involvement of endogenous prostaglandins in regulation of uterine spontaneous contraction was investigated using mechanical, immunohistochemical and biochemical approaches. Immunohistochemical study and Western blot analysis for immunoreactive cyclooxygenase (COX) indicated that COX-1 but not COX-2 was expressed predominantly in the myometrium of non-pregnant pigs in a muscle layer-dependent manner (longitudinal muscle>circular muscle). Pretreatment of uterine strips with indomethacin and selective COX-1 inhibitors (SC-560 and FR122047) significantly reduced both the amplitude and frequency of spontaneous contraction in the longitudinal muscle, but inhibition by COX inhibitors was negligible in the circular muscle. On the other hand, CAY10404, a COX-2 inhibitor, did not change the spontaneous contraction in either of the muscle layers. Pretreatment with SC-560 reduced myometrial PGF(2alpha) and PGE(2) levels. Contractile FP and EP(3) receptors were expressed in a muscle layer-dependent manner (longitudinal muscle>circular muscle), similar to the expression pattern of COX-1. In conclusion, endogenous prostaglandins produced by COX-1 regulate spontaneous contractile activity of non-pregnant porcine uterine longitudinal muscle selectively due to the heterogeneous expression of contractile prostanoid receptors and COX-1.     

Isolation, partial purification and pharmacodynamics of a slow contractile substance in the venom sac extract of the wasp Vespa cincta Fabr

The venom of V. cincta contains acetylcholine (ACh), histamine and 5-hydroxytryptamine (5-HT). Blockers of these agonists did not block completely the hypotensive and smooth muscle contractile activity of venom. On smooth muscle, there was a residual slow contraction. The active substance which produced this slow contraction was separated by solvent extraction, gel filtration and TLC. The purified material (which has been provisionally designated "Vecikinin") lowered cat, rat and guinea pig blood pressure, increased amplitude of cardiac contraction, and increased capillary permeability. Vecikinin contracted several smooth muscle preparations (rat uterus, rat ascending colon, guinea pig ileum, guinea pig colon and rat ileum), while relaxing rat duodenum. Its contractile activity was not lost on boiling, but acid or alkali-boiling reduced its contractile activity. It was inactivated on incubation with chymotrypsin and carboxypeptidase but not with trypsin, pepsin or leucine aminopeptidase. It is a peptide, appears to be of low molecular weight, and could be distinguished from substance P, angiotensin, bradykinin and hornet or wasp kinin.     

Generation of spiking activity in circular muscle cells of the canine colon

Spontaneous and current-induced electrical activity was recorded intracellularly to resolve the controversy whether or not the circular muscle layer of the colon generates spiking activity. Particularly in the first hour after mounting the tissue in the organ bath, spikes were recorded at both the submucosal and the myenteric plexus side of the muscle layer. Spikes were seen as part of the slow wave upstroke in the submucosal surface cells, and spikes occurred both at the upstroke potential and superimposed on the plateau potential in myenteric plexus surface cells. Spikes increased the force of contraction. The study supports earlier claims using extracellular recording techniques that circular muscle cells generate spiking activity, particularly in the presence of depolarizing stimuli, and that spikes contribute to contractile activity.     

Effect of contraction frequency on the contractile and noncontractile phases of muscle venous blood flow.

The purpose of this study was to test the hypothesis that increasing muscle contraction frequency, which alters the duty cycle and metabolic rate, would increase the contribution of the contractile phase to mean venous blood flow in isolated skeletal muscle during rhythmic contractions. Canine gastrocnemius muscle (n = 5) was isolated, and 3-min stimulation periods of isometric, tetanic contractions were elicited sequentially at rates of 0.25, 0.33, and 0.5 contractions/s. The O2 uptake, tension-time integral, and mean venous blood flow increased significantly (P < 0.05) with each contraction frequency. Venous blood flow during both the contractile (106 +/- 6, 139 +/- 8, and 145 +/- 8 ml x 100 g-1 x min-1) and noncontractile phases (64 +/- 3, 78 +/- 4, and 91 +/- 5 ml x 100 g-1 x min-1) increased with contraction frequency. Although developed force and duration of the contractile phase were never significantly different for a single contraction during the three contraction frequencies, the amount of blood expelled from the muscle during an individual contraction increased significantly with contraction frequency (0.24 +/- 0.03, 0.32 +/- 0.02, and 0.36 +/- 0.03 ml x N-1 x min-1, respectively). This increased blood expulsion per contraction, coupled with the decreased time in the noncontractile phase as contraction frequency increased, resulted in the contractile phase contribution to mean venous blood flow becoming significantly greater (21 +/- 4, 30 +/- 4, and 38 +/- 6%) as contraction frequency increased. These results demonstrate that the percent contribution of the muscle contractile phase to mean venous blood flow becomes significantly greater as contraction frequency (and thereby duty cycle and metabolic rate) increases and that this is in part due to increased blood expulsion per contraction.     

Enteric opioid neurons modulate the basal tone of the isolated puppy ileum

We studied the role of enteric opioid neurons in the spontaneous motility of the longitudinal muscle in the isolated puppy ileum. Regular fluctuations in tone that rose above and returned to the basal level occurred at an interval of 4.7 +/- 0.3 min. Naloxone (10(-8) and 10(-7) M) reduced the spontaneous tonic contraction by 42.6 +/- 11.6% (p less than 0.02) and 77.0 +/- 3.6% (p less than 0.001), respectively. Tetrodotoxin (3.1 X 10(-7) M) and atropine (10(-7) M) terminated the fluctuations. Met- and Leu-enkephalins (10(-9)-10(-8) M) caused tonic contraction which was abolished by tetrodotoxin and atropine. The contractile response produced by transmural electrical stimulation was reduced by naloxone (10(-7) M). This response was also abolished by atropine and tetrodotoxin. These results suggest that enteric opioid neurons are spontaneously active and might operate, at least in part, to raise the basal tone of the longitudinal muscle in the puppy ileum through a cholinergic excitatory mechanism.