Neural control of gastro-intestinal motility; Events following receptor activation

The motility of the mammalian gastro-intestinal tract is regulated by a number of neuronal systems. The postganglionic, intramural parts of these systems are characterized by their neurotransmitters as cholinergic, adrenergic and purinergic nerves, respectively. The transmitter acetylcholine evokes contractions of the smooth muscle cell, whereas both noradrenalin and the transmitter from the purinergic nerves, presumably ATP, induce relaxations of smooth muscles. Agents and actions which influence the neurohumoral transmission and the mechanisms underlying the effects induced by the transmitters in smooth muscle cells are reviewed.     

Adrenergic and peptidergic innervation of the trachealis muscle in the normal horse: a preliminary report

The tone of respiratory smooth muscle is largely determined by the input from autonomic nerves. The distribution of adrenergic and selected nonadrenergic, non-cholinergic (NANC) nerves in the normal equine trachealis muscle was investigated using immunohistochemistry. The smooth muscle of the trachealis was found to contain numerous nerves immunoreactive for an enzymatic marker of adrenergic nerves, as well as many nerves immunoreactive for a putative NANC neurotransmitter, peptide histidine isoleucine, a potent bronchodilator. The tissues surrounding the respiratory smooth muscle contained numerous nerves immunoreactive for the neuropeptides substance P and calcitonin gene-related peptide, which can cause marked vasodilation and bronchoconstriction. The complex innervation of the equine trachea should be kept in mind when interpreting the results of physiological experiments.     

Neurogenic and Myogenic Responses in Isolated Smooth Muscle of The Bovine Teat

Strips of bovine teat smooth muscle were studied. Their innervation was investigated with analytical pharmacology on field stimulated preparations. The content of noradrenaline in the teat wall and the sphincter region was determined. The main results were: teat smooth muscle has properties of single unit smooth muscle i.e. automaticity and active response to stretch. Especially the sphincter region is inclined to rapid phasic rhythmicity. The teat smooth muscle receives excitatory adrenergic innervation. The noradrenaline released from the nerves preferentially activates α-adrenoceptors, while β-adrenoceptors essentially are noninnervated and humoral. The sphincter region has a higher concentration of noradrenaline and a more intimate neuro-effector arrangement. This is probably of functional significance. Moderate chilling increases the response to adrenergic nerve stimulation while it is decreased at higher temperatures. This has most likely physiological importance. The results are discussed with respect to in vivo findings and it is concluded that the bovine teat smooth muscle with its excitatory adrenergic nerves forms a normally very efficient neuro-effector system with a particular strength in the sphincter around the streak canal.     

Cisapride Stimulates Contraction of Idiopathic Megacolonic Smooth Muscle in Cats

We have previously shown that cisapride, a substituted piperidinyl benzamide, stimulates contraction of healthy feline colonic smooth muscle. The purpose of the present investigation was to determine the effect of cisapride on feline idiopathic megacolonic smooth muscle function. Longitudinal smooth muscle strips from ascending and descending colon were obtained from cats with idiopathic megacolon, suspended in a 1.5 mM Ca²⁺-HEPES buffer solution (37°C, 100% O₂, pH 7.4), attached to isometric force transducers, and stretched to optimal muscle length (L₀). Control responses were obtained at each muscle site with acetylcholine (10–⁸ to 10 ⁴ M), substance P (10–¹¹ to 10^-7 M), or potassium chloride (10 to 80 mM). Muscles were then stimulated with cumulative (10–⁹ to 10–⁶ M) doses of cisapride in the absence or presence of tetrodotoxin (10–⁶ M) and atropine (10–⁶ M), or in a 0 calcium HEPES buffer solution. In cats with idiopathic megacolon, cisapride stimulated contractions of longitudinal smooth muscle from both the ascending and the descending colon. Cisapride-induced contractions were similar in magnitude to those induced by substance P and acetylcholine in the ascending colon, but were less than those observed in the descending colon. Cisapride-induced contractions in megacolonic smooth muscle were only partially inhibited by tetrodotoxin and atropine, but were virtually abolished by removal of extracellular calcium. We concluded that cisapride-induced contractions of feline megacolonic smooth muscle are largely smooth muscle mediated and dependent on influx of extracellular calcium. Cisapride-induced contractions in megacolonic smooth muscle are only partially dependent on enteric cholinergic nerves. Thus, cisapride may be useful in the treatment of cats with idiopathic megacolon.     

5-Hydroxytryptamine potentiates contraction mediated by the intramural cholinergic nerve in the longitudinal smooth muscle of the ruminant forestomach

The effects of 5-hydroxytryptamine (5-HT) on the longitudinal smooth muscle from the rumen and reticulum of the bovine forestomach were investigated. 5-HT (0.25–490 μM) caused a contraction and a relaxation of the ruminal strips while it produced only an excitatory effect on the reticular strips. These effects were not affected by tetrodotoxin, hexamethonium, atropine or morphine, but were blocked by methysergide, LSD-25 or phenoxybenzamine. 5-HT potentiated the contraction evoked by stimulation of the intramural cholinergic nerves but did not show any effect on the relaxation produced by the non-adrenergic inhibitory nerves' excitation. The 5-HT-induced potentiation was not affected by morphine, LSD-25, methysergide and hexamethonium or high concentration of nicotine. Nicotine and dimethylphenylpiperazinium also caused a transient augmentation of the nerve-mediated contraction, but these effects were abolished by the competitive ganglionic blockers. The evoked contraction was depressed in high-Mg²⁺ solution, but this depression was antagonized partly by 5-HT. The affinity of the cholinomimetics to post-synaptic muscarinic receptor was not affected by 5-HT. It is concluded that contractions or relaxations of bovine forestomach strips induced by 5-HT are mediated through activation of D-receptors in the smooth muscle, and the 5-HT-induced potentiation of the evoked contraction may be elicited through presynaptic neural effects of 5-HT on the cholinergic nerves.     

Influence of motor activities on the release of transmitter quanta from motor nerve terminals in mice.

We investigated the effects of motor activities on transmitter release in mouse nerve-muscle preparations of the diaphragm muscle (DPH), extensor digitorum longus muscle (EDL), and soleus muscle (SOL). Mice were divided into a control group, a motor-restricted (RST) group, and a motor-compelled (CMP) group. The quantal content (m) of endplate potentials was measured intracellularly. In DPH the motor activity was unaffected. In the CMP group the m value of the EDL group increased with increases in the cooperativity of Ca2+ in transmitter release. Compared with the CMP group, the SOL of the RST group had a smaller m value with increases in the cooperativity of Ca2+ in transmitter release. These results suggest that motor activities can influence neuromuscular activity specific to different systems, however, the motor compulsion specifically activated the function of EDL and the motor restriction activated the function of SOL, and these effects might lead to altered activity of the release of transmitter quanta in motor nerve terminals of mice.     

In vitro effects of erythromycin, lidocaine, and metoclopramide on smooth muscle from the pyloric antrum, proximal portion of the duodenum, and middle portion of the jejunum of horses

OBJECTIVE: To evaluate effects of erythromycin, lidocaine, and metoclopramide on smooth muscle of the pyloric antrum (PA), proximal portion of the duodenum (PD), and middle portion of the jejunum (MJ) of horses. Sample Population-Strips of smooth muscle from 7 horses. PROCEDURE: Isolated muscle strips were suspended in a bath and attached to isometric force transducers. Once stable spontaneous contractions were observed, agents were added. Isometric stress responses were compared with the amplitude of spontaneous contractions. RESULTS: A single dose of erythromycin to the PA increased contractile amplitude (CA) for the longitudinal smooth muscle (mean +/- SEM, 76+/-16 g/cm2) but decreased CA for circular smooth muscle (-79+/-23 g/cm2). The inhibitory effect was decreased by tetrodotoxin, N(G)-nitro-L-arginine methyl ester, and a vasoactive intestinal peptide antagonist. Erythromycin increased CA for the MJ, which was maximal at 10(-4)M (171+/-36 g/cm2). Lidocaine increased CA for the PD, which was maximal at 10(-4) M (60+/-5 g/cm2). Metoclopramide increased the CA, which was maximal at 10(-4) M for the PA (75+/-26 g/cm2), PD (279+/-33 g/cm2), and MJ (456+/-59 g/cm2). CONCLUSIONS: Regional differences in responses to erythromycin, lidocaine, and metoclopramide were evident in the gastrointestinal tract of horses. Metoclopramide increased CA in all tissues used, whereas erythromycin inhibited CA in circular smooth muscle but stimulated CA in longitudinal smooth muscle from the PA. Inhibition is caused by stimulation of inhibitory nerves and is mediated, in part, by nitric oxide and vasoactive intestinal peptide.     

Response of equine airway smooth muscle to acetylcholine and electrical stimulation in vitro

Smooth muscle strips from the midcervical portion of the trachea and bronchial smooth muscle strips from third-generation airways of horses were placed in tissue baths, and isometric contractile force was measured. Active force was measured in response to electrical stimulation and exogenous acetylcholine. Square-wave electrical stimuli were applied at various voltages (10, 12, 15, 18, 20, 25 V), frequencies (3, 5, 10, 15, 20, 25, 30 Hz), and pulse durations (0.2, 0.5, 1.0, 1.5, 2.0 ms). Isometric contractile force increased as voltage, frequency, and pulse duration increased. Maximal contractile response to electrical stimulation was obtained at 18 V, 25 Hz, and 0.5 ms. Atropine (10(-6)M) or tetrodotoxin (3 x 10(-6)M) blocked the contraction, indicating that the contractile response was attributable to the release of neurotransmitter from cholinergic nerves. Cumulative concentration-response curves to acetylcholine (10(-9)M through 10(-4)M) were determined. Isometric contractile force increased as acetylcholine concentration increased. There was a significant (P less than 0.05) difference in the 50% effective dose for acetylcholine in tracheal smooth muscle and bronchial smooth muscle. The mean (+/- SD) contractile response to maximal electrical stimulus was 89% (+/- 7.4%) of that in response to 10(-4)M acetylcholine in tracheal smooth muscle and was 68% (+/- 10.4%) of the response to 10(-4)M acetylcholine in bronchial smooth muscle.     

应激抑制妊娠母兔子宫平滑肌收缩时间的初探

催产素能使妊娠 1 5d后的母兔子宫出现收缩反应。但在妊娠母兔受到强应激刺激后立即注射催产素子宫不发生收缩。说明当应激达到一定强度时 ,母兔因紧张所分泌的肾上腺素足以封闭子宫平滑肌上的催产素受体 ,从而使催产素在一定的时间内不能使子宫收缩     

A progressive axonopathy of Boxer dogs affecting the central and peripheral nervous systems

Five Boxer dogs with a new neurological disease are described. There is a progressive ataxia and weakness, initially in the hind-legs but later involving forelegs. The onset of signs is usually around 6 months of age although an older dog was affected. Proprioceptive function, muscle tone and tendon reflexes are diminished or absent while pedal reflexes and pain sensation are preserved. Muscle atrophy is minimal. The peripheral nerves are depleted in myelinated fibres. Nerve roots and, to a lesser extent, peripheral nerves show demyelination/remyelination changes and in the cervical ventral roots in particular, regenerative clusters are present. Forelimb muscles show evidence of re-innervation. In the CNS numerous swollen axons are present. The dorsolateral and ventral columns of the spinal cord are affected throughout their length. In the brain a number of nuclei and tracts, particularly in the lower brain stem, are affected and the superior olives are severely involved. The cerebral cortex and subcortical white matter appear normal. Various portions of the auditory and optic pathways are affected pathologically as are a number of cranial nerves. The axonal swellings contain a number of organelles including smooth endoplasmic reticulum, disordered neurofilaments, vesicles and mitochondria. The disease may be specific to Boxer dogs and may be inherited.     

In vitro effects of lidocaine on the contractility of equine jejunal smooth muscle challenged by ischaemia‐reperfusion injury

Reasons for performing study: Post operative ileus (POI) in horses is a severe complication after colic surgery. A commonly used prokinetic drug is lidocaine, which has been shown to have stimulatory effects on intestinal motility. The cellular mechanisms through which lidocaine affects smooth muscle activity are not yet known. Objectives: To examine the effects of lidocaine on smooth muscle in vitro and identify mechanisms by which it may affect the contractility of intestinal smooth muscle. Hypothesis: Ischaemia and reperfusion associated with intestinal strangulation can cause smooth muscle injury. Consequently, muscle cell functionality and contractile performance is decreased. Lidocaine can improve basic cell functions and thereby muscle cell contractility especially in ischaemia‐reperfusion‐challenged smooth muscle. Methods: To examine the effects of lidocaine on smooth muscle function directly, isometric force performance was measured in vitro in noninjured and in vivo ischaemia‐reperfusion injured smooth muscle tissues. Dose‐dependent response of lidocaine was measured in both samples. To assess membrane permeability as a marker of basic cell function, release of creatine kinase (CK) was measured by in vitro incubations. Results: Lidocaine‐stimulated contractility of ischaemia‐reperfusion injured smooth muscle was more pronounced than that of noninjured smooth muscle. A 3‐phasic dose‐dependency was observed with an initial recovery of contractility especially in ischaemia‐reperfusion injured smooth muscle followed by a plateau phase where contractility was maintained over a broad concentration range. CK release was decreased by lidocaine. Conclusion: Lidocaine may improve smooth muscle contractility and basic cell function by cellular repair mechanisms which are still unknown. Improving contractility of smooth muscle after ischaemia‐reperfusion injury is essential in recovery of propulsive intestinal motility. Potential relevance: Characterisation of the cellular mechanisms of effects of lidocaine, especially on ischaemia‐reperfusion injured smooth muscle, may lead to improved treatment strategies for horses with POI.     

Is the Rhesus Monkey (Macaca mulatta) Comparable to Humans? Histomorphology of the Sphincteric Musculature of the Lower Urinary Tract Including 3D‐reconstruction

The physiology of the muscle systems of the human lower urinary tract is still not known in detail. To study the functional basics of this complex organ system, experiments are often performed in animal models including rhesus monkeys. To apply the results of animal model studies to the humans, a clear knowledge of the comparative anatomy of both species is necessary. However, detailed comparative studies of the lower urinary tract of the rhesus monkey and the humans are lacking. Accordingly, a detailed study on the sphincteric musculature of the lower urinary tract of the rhesus monkey was performed in order to demonstrate anatomical correspondences and differences between both species. The lower urinary tract anatomy was investigated in 18 male and female rhesus monkeys (Macaca mulatta) by serial sections. Immunohistochemical staining methods were used to differentiate striated and smooth musculature. Three-dimensional reconstructions were performed in order to demonstrate the topographical anatomy of the different muscle systems. In both man and male rhesus monkeys, a urethral sphincter muscle exists independent of the pelvic floor musculature, with a smooth and a striated muscular part. A urinary diaphragm (diaphragma urogenitale) does neither exist in the rhesus monkey nor in the human. In contrast to women, a striated muscle encircles the urethra and vagina together in the female rhesus monkey. A vesical sphincter muscle, found in the human bladder outlet, does not exist in the rhesus monkey.     

中药产复康对大鼠离体子宫平滑肌收缩功能的影响

为了观察中药制剂产复康对离体子宫平滑肌的收缩功能,测定了产复康对大鼠离体子宫平滑肌收缩功能和催产素引起的大鼠离体子宫平滑肌收缩功能的影响。结果发现低剂量产复康可以促进大鼠离体子宫平滑肌的收缩功能,提高子宫的活动性,高剂量则对子宫呈抑制作用;产复康对催产素引起的大鼠离体子宫平滑肌收缩功能具有抑制作用,且呈剂量-效应关系。产复康对大鼠离体子宫具有双向调节作用。     

奶牛皱胃平滑肌细胞体外培养方法的建立

平滑肌细胞是胃动力的基本单位,为研究奶牛皱胃弛缓的发病机理,本研究建立了奶牛皱胃平滑肌细胞的体外培养方法。手术取奶牛皱胃组织,镜下分离出肌层,运用酶消化法分离皱胃平滑肌细胞并进行体外培养,在倒置显微镜下观察细胞的生长状态,用免疫组化染色方法进行平滑肌细胞鉴定。结果显示细胞生长状态良好,呈现平滑肌细胞特征性的"峰-谷"状结构,免疫组化染色显示细胞浆内平滑肌肌动蛋白呈阳性表达,表明该细胞为平滑肌细胞,可用于后续的试验研究。     

Tyrosine hydroxylase- and neuropeptides-immunoreactive nerves in canine trachea

OBJECTIVE: To determine distribution of catecholaminergic and peptidergic nerve fibers in canine tracheas by use of immunohistochemistry. SAMPLE POPULATION: 10 tracheas collected from healthy adult dogs after euthanasia. PROCEDURE: Structure of the nerve network and distribution of tyrosine hydroxylase (TH)- and 6 types of neuropeptide-containing nerves in canine tracheas were immunohistochemically studied, using neurochemical markers. RESULTS: Intraepithelial free nerve endings with immunoreactivity for calcitonin gene-related peptide (CGRP) and substance P (SP) were observed.Tyrosine hydroxylase-, SP-, vasoactive intestinal peptide (VIP)-, and galanin (GAL)-immunoreactive nerve fibers were observed within and around the submucosal seromucous gland. In the smooth muscle layer, numerous TH- and GAL-immunoreactive nerve fibers, a moderate number of VIP- and neuropeptide Y (NPY)-immunoreactive nerve fibers, and a few SP- and methionine enkephalin (ENK)-immunoreactive nerve fibers were observed. Numerous nerve cell bodies with VIP and GAL immunoreactivity and a few with SP ENK, and NPY immunoreactivity were observed. Many TH-immunoreactive fibers were arranged in a meshwork around blood vessels. Nerves with CGRP-, SP-, VIP-, GAL-, ENK-, and NPY-immunoreactivity were also observed around blood vessels. CONCLUSIONS: Complex innervation, including catecholamine- and neuropeptide-containing nerves, which may be related to regulation of muscle contraction and glandular secretion, are found in canine tracheas.     

Autonomic innervation of the airways

Summary

A review is given of the literature concerning the autonomic innervation of airway smooth muscle. The cholinergic, adrenergic and non‐cholinergic non‐adrenergic (NANC) systems in humans and several animal species are discussed. The diagnostic and therapeutic possibilities and limitations of new receptor specific agonists and antagonists are also discussed.     

紫菀乙醇提取物对豚鼠离体气管平滑肌收缩功能的影响

本试验旨在研究紫菀乙醇提物对豚鼠离体气管平滑肌收缩功能的影响。采用离体气管恒温灌流的方法,将豚鼠气管制成气管螺旋条,通过 BL-420F 生物机能实验系统测定其张力的变化,观察紫菀乙醇提物对离体气管平滑肌静息状态下的舒张作用,以及在氯化乙酰胆碱(Ach)、磷酸组胺(His)、CaCl₂条件下和无钙下Ach诱导细胞内钙释放和外钙内流所致两种收缩条件下对离体气管平滑肌张力变化的影响。试验结果显示,低浓度紫菀乙醇提取物(0.002~0.008 g/mL)对静息状态下豚鼠离体气管平滑肌具有一定的收缩作用,高浓度(0.008~0.196 g/mL)时具有舒张作用;低、中、高3个剂量组均可抑制Ach、His和CaCl₂引起的气管平滑肌收缩强度,使各致痉剂的量效曲线非平行右移并降低最大效应。紫菀乙醇提取物对His的抑制作用强度最大,其次为Ach,最后是CaCl₂,且均呈剂量依赖性。以上结果表明紫菀乙醇提物具有舒张气管平滑肌的作用,其机制可能与抑制豚鼠气管平滑肌M受体、H₁受体和阻断Ca²⁺通道从而抑制细胞Ca²⁺内流有关。     

Effects of Alcohol Extract of Aster tataricus L.f.on the Contraction of Guinea Pig Tracheal Smooth Muscle in vitro

The aim of the experiment was to study the effect of alcohol extract of Aster tataricus L.f.on the contraction of isolated guinea pig tracheal smooth muscle.The guinea pig tracheal smooth muscle was prepared by isolated tracheal thermostatic perfusion method.The effects of Aster tataricus L.f.on tracheal smooth muscle under the resting state and contraction stimulated by acetylcholine (Ach),histamine (His),CaCl₂,Ca²⁺ release and influx in cells without calcium were measured by BL-420F biological function experiment systems.The results showed that Aster tataricus L.f.could induce the contraction of isolated trachea at resting state at a low concentrations of 0.002 to 0.008 g/mL and exerted a relaxation on the isolated trachea when at the higher concentrations of 0.008 to 0.196 g/mL.All of the three concentrations of Aster tataricus L.f.could inhibit the tension stimulated by Ach,His and CaCl₂ which were antispasmodic agents,led to the non-parallelled rightward moving of cumulative concentration-response curve and the decline of maximal responses.The suppressive effect on His was the strongest,followed by Ach and CaCl₂,and the suppression could counteract the contraction induced by extracellular Ca²⁺influx significantly,and they were all expressed concentration-dependent manner.These results indicated the alcohol extract of Aster tataricus L.f.could relax tracheal smooth muscle by acting on M-receptor and histamine receptor and blocking Ca²⁺ passage,thus inhibiting extracellular Ca²⁺influx.     

Regulatory peptides

The peptidergic innervation of the gastrointestinal tract was clearly affected in these six cases of Feline Dysautonomia. Most notably there was a striking depletion of VIP-immunoreactivity. However, the lesions in Feline Dysautonomia are not selective for neurons containing this particular peptide, since a reduction in substance P- and Met-enkephalin-immunoreactivity was also found. Depletion of these three neuropeptides does not correlate well with the clinical finding of gastrointestinal atony, since only substance P has excitatory actions on gut smooth muscle (Iversen, 1982). Both VIP and enkephalin have inhibitory actions, acting directly on smooth muscle or via the myenteric plexus respectively (Fahrenkrug and Emson, 1982; North and Egan, 1982). However, as there was no specificity in the distribution of lesions, at least in terms of peptidergic nerves, it is probable that other types of nerves in the intrinsic and extrinsic innervation of the gut are affected, and contribute to the loss of tone. The depletion of neuropeptide immunoreactivity does not distinguish between a loss of peptidergic neurons and a loss of peptide content. However, in routinely stained sections there was little evidence of neuronal loss: only a small number of abnormal, apparently degenerating, nerve cell bodies was seen in the enteric plexuses. Thus, it is possible that levels of immunoreactive materials are reduced in peptidergic nerves. At the ultrastructural level, Griffiths and Sharp (1983) have found disorganized rough endoplasmic reticulum and a depletion of ribosomes in neurons from affected cats, and it is possible that impaired peptide synthesis may contribute to the observed reduction in immunoreactivity. In contrast to the findings in the peptidergic innervation, the peptide-containing endocrine cells appeared to be unaffected. At least, glucagon-and somatostatin-IR cells in the colon were not depleted in number. Thus, in Feline Dysautonomia, while there was little evidence of neuronal lesions in the gastrointestinal tract on routine histological examination, there was a marked loss of neuropeptide immunoreactivity. Neuropeptide immunohistochemistry would appear, therefore, to be more sensitive in detecting neuronal abnormalities in this disease. At present however, it cannot be used as a diagnostic tool for Feline Dysautonomia, as regulatory peptides are known to be involved in several other diseases of the mammalian gastrointestinal tract (Vaillant and others, 1982; Bloom and Polak, 1982). Further studies are required to distinguish the enteric neuropeptide abnormalities in Feline Dysautonomia from those occurring in other disorders affecting the feline gut.     

Smooth Muscle Cells of the Bovine Cervical Stroma may have a Secretory, rather than a Contractile Function during Parturition

The bovine cervix contains a large amount of smooth muscle cells distributed over an outer muscular layer and within a stromal layer. The stromal layer exhibits no electromyographic (EMG) activity at parturition. This leads to the question whether the stromal smooth muscle cells of the bovine cervix are prepared to contract with parturition, or whether they have another function. To this end, cervical biopsies were repeatedly taken from 10 pregnant cows at day-185 and -275 of gestation, at spontaneous, uncomplicated calving and at 30 days after calving. The smooth muscle bundles of the stroma were immunohistochemically analysed (n = 5) with regard to their integrity and cellular density, and the degree of staining for connexin-43, smooth muscle actin α (SMA), desmin and vimentin. Additionally, the mRNA expression for connexin-43, SMA, desmin and vimentin was determined with RT-PCR (n = 5). The smooth muscle tissue was arranged in bundles, also at parturition. However, the cellular density of these bundles and the SMA mRNA expression were decreased at parturition. Additionally, the SMA staining and connexin-43 expression and staining remained constant during pregnancy and at parturition. This might indicate that stromal smooth muscle cells are not prepared to contract with parturition, in contrast to the myometrial smooth muscle cells. The smooth muscle cells, stained for SMA, also expressed vimentin, and the proportion of co-expression was increased at day-275 of pregnancy. This suggests that the stromal smooth muscle cells predominantly have a secretory function in cows.