Contraction Of Circular Smooth Muscle Research Articles

Circular muscle contraction, messenger signalling and localization of binding sites for neurokinin A in human sigmoid colon.

1. Neurokinin (NK)A is the endogenous ligand for the tachykinin NK2 receptor. In the present study, tachykinins and selective receptor agonists were tested as contractile agonists in human colon circular muscle and [125I]-NKA was used to localize binding sites in human colon. 2. In strips of circular muscle, removal of mucosa and submucosa significantly (P < 0.05) increased the potency and the maximum response achieved by NKA. 3. The rank order of potency of tachykinin and selective receptor agonists in contracting circular muscle strips was NKA > or = [Lys5,MeLeu9,Nle10]NKA(4-10) > or = neuropeptide (NP)gamma > or = [betaAla8]NKA(4-10) >> NKB > substance P (SP) >> senktide approximate to [Pro9]SP. 4. Specific binding sites for [125I]-NKA were densely localized over circular muscle and muscularis mucosae. Weak specific binding was seen on longitudinal muscle and taenia coli, whereas no binding sites were seen on mucosa, ganglia or blood vessels. 5. In circular muscle, the selective NK2 receptor agonist [LysS,MeLeu9,Nle10]NKA(4-10) produced weak increases (maximum 37%) in inositol monophosphate formation with a pD2 of 6.8+/-0.51 (n = 3). Carbachol (100 micromol/L) was also a weak stimulant (maximum 45%). These agonists were over 10-fold more efficacious in stimulation of inositol monophosphate in rat urinary bladder. 6. In conclusion, [125I]-NKA binding sites localized on human colon circular muscle were characterized as NK2 receptors. Functionally, the tachykinin NK2 receptor is mediating circular smooth muscle contraction. Although the human NK2 receptor is coupled to the phosphatidylinositol pathway, other second messenger mechanisms may also operate in this tissue.

Gq-linked NK2 receptors mediate neurally induced contraction of human sigmoid circular smooth muscle

Because tachykinins have been identified as neurotransmitters in the guinea pig colon and human ileum, we examined a possible role of tachykinin receptors and neurokinin (NK) A in neurally induced contraction of human sigmoid colon circular muscle. Muscle strips were stimulated electrically for 10 seconds. Single cells were isolated by enzymatic digestion and permeabilized by saponin. [(35)S]GTPgammaS binding was assayed with or without NKA for 5 minutes. Intracellular Ca(2+) was measured using Fura 2. In the presence of 100 micromol/L L-NNA, 100 micromol/L atropine did not affect electrical field stimulation (EFS)-induced contraction. A peptide NK(2)-receptor antagonist (NK-2ra) but not an NK(1) antagonist FK888 (1 micromol/L) eliminated EFS-induced contraction. NKA-induced contraction in muscle strips and single cells was virtually abolished by NK-2ra, but not by FK888. In permeabilized cells, contraction was blocked by Gq-protein antibodies, but not by other G-protein antibodies, suggesting that NKA activates Gq, which was confirmed by a [(35)S]GTPgammaS binding assay. NKA-induced contraction and increase in cytosolic Ca(2+) were abolished by depletion of intracellular Ca(2+) stores. Tachykinins may be the main excitatory neurotransmitters in human sigmoid circular muscle. NKA activates Gq-linked NK(2) receptors, which cause Ca(2+) release, followed by contraction.

Thromboxane A 2-induced contraction of cat esophageal and lower esophageal sphincter circular smooth muscle

Thromboxane A 2-induced contraction of cat esophageal and lower esophageal sphincter circular smooth muscle

Downregulation of Galphaq-11 protein expression in guinea pig antral and colonic circular muscle during pregnancy.

Pregnancy has an inhibitory effect on motility of the gastrointestinal tract. The present study was designed to examine the mechanisms responsible for antral and colonic hypomotility in pregnant guinea pigs. Circular smooth muscle cells from the antrum and left colon were isolated by enzymatic digestion with collagenase from pregnant and nonpregnant guinea pigs. Contractile responses to agonists were expressed as percent shortening from resting cell length. The function of G proteins in antral and colonic circular smooth muscle was assessed by [35S]guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) binding induced by CCK-8 and G protein quantitation. The contraction of antral and colonic circular smooth muscle from pregnant guinea pigs was reduced in response to CCK-8 and to GTPgammaS but was normal in response to KCl and D-myo-inositol 1,4,5-trisphosphate compared with nonpregnant animals. The stimulation of [35S]GTPgammaS binding to Galphaq-11 induced by 1 microM CCK-8 was significantly lower in antral and colonic circular smooth muscle from pregnant guinea pigs than that in controls. Furthermore, Western blot analysis showed a decreased Galphaq-11 and an increased Gsalpha protein content in both tissues during pregnancy. It is concluded that pregnancy appears to impair gastrointestinal circular smooth muscle contractility by downregulating G proteins such as Galphaq-11 protein, which mediates muscle contraction, and upregulating Gsalpha protein, which mediates muscle relaxation.

Characterization of 5-Hydroxytryptamine Receptors Mediating Circular Smooth Muscle Contraction in the Human Umbilical Artery

The study was performed to characterize pharmacologically the contractile 5-hydroxytryptamine (5-HT) receptors in the circular smooth muscle of the isolated human umbilical artery. Effects of agonists and antagonists for different 5-HT receptor subtypes were studied in intact endothelium vessel segments. All agonists induced concentration-dependent circular smooth muscle contractions. The potency was in declining order 5-HT > α-methyl-5-HT > sumatriptan ≥ 2-methyl-5-HT. The effects of 5-HT and α-methyl-5-HT were antagonized by ketanserin, as well as methiothepin. The contractile effect of sumatriptan was antagonized by methiothepin but not by ketanserin. The 5-HT₃ receptor antagonist, MDL 72222, did not affect the contraction by any of the agonists, including 2-methyl-5-HT. It is concluded that the 5-HT-induced contraction in the circular smooth muscle of the human umbilical artery seems to be mediated by a mixed population of 5-HT₁-like receptors and 5-HT₂ receptors.

Leukotriene D 4–induced contraction of cat esophageal and lower esophageal sphincter circular smooth muscle

In esophageal circular muscle, acetylcholine activates phosphatidylcholine-specific phospholipases C and D and phospholipase A2, producing diacylglycerol and arachidonic acid, which cause contraction by interacting synergistically to activate protein kinase C. In a model of acute esophagitis, leukotriene D4 (LTD4) contributes to acetylcholine-induced contraction. We examined intracellular signaling in LTD4-induced contraction. Esophageal and lower esophageal sphincter (LES) cells, isolated by enzymatic digestion, were contracted by LTD4 in the absence or presence of inhibitors. Permeabilization by saponin allowed use of G-protein antibodies and heparin. Esophageal contraction was inhibited by pertussis toxin, Gi3 antibodies, D609 (phosphatidylcholine-specific phospholipase C inhibitor), propranolol (phospholipase D pathway inhibitor), and chelerythrine (protein kinase C antagonist) but not W7 (calmodulin antagonist). LES contraction was unaffected by pertussis toxin. It was inhibited by Gq antibodies, U-73122 (phosphatidylinositol-specific phospholipase C inhibitor), heparin (inositol 1,4,5-trisphosphate inhibitor), and W7 and reduced by D609. In the esophagus, LTD4 activates a protein kinase C-dependent pathway through pertussis toxin-sensitive Gi3 proteins and phosphatidylcholine-specific phospholipase. In the LES, LTD4 activates a calmodulin-dependent pathway through pertussis toxin-insensitive Gq proteins and phosphatidylinositol-specific phospholipase C. The intracellular pathways activated by LTD4 in the esophagus and the LES are similar to those activated by acetylcholine and other agonists.

MAP kinase mediates PKC-dependent contraction of cat LES and esophageal circular smooth muscle

MAP kinase mediates PKC-dependent contraction of cat LES and esophageal circular smooth muscle

Delta-opioid receptor mRNA expression and immunohistochemical localization in porcine ileum.

Opiates have potent antidiarrheal actions that are mediated in part by delta-opioid receptors (DOR). We examined DOR localization within subregions of porcine ileum, a tissue analogous to human small bowel. A partial cDNA sequence for porcine DOR was obtained after reverse transcription-polymerase chain reaction cloning of forebrain RNA; it encoded the end of transmembrane domain 1 through the beginning of transmembrane domain 7 and exhibited 93% nucleotide identity with human DOR. Positive signals for DOR mRNA were found in all subregions of the porcine ileal wall. With an antiserum recognizing an N-terminal epitope in murine DOR, DOR-like immunoreactivity was localized in neurons within myenteric and submucous ganglia, longitudinal and circular smooth muscle, and villous lamina propria. The DOR agonist [D-Ser2, Leu5, Thr6]enkephalin (DSLET) attenuated circular smooth muscle contractions in porcine ileum that were evoked by electrical stimulation of myenteric cholinergic neurons. These results are consistent with previous reports of the DOR-mediated neuromodulation that underlies the antipropulsive and antisecretory effects of opioids in the intestinal tract.

Leukotrienes in acetylcholine-induced contraction of esophageal circular smooth muscle in experimental esophagitis

Phospholipase A2 (PLA2) participates in acetylcholine (ACh)-induced contraction of esophageal circular smooth muscle. Because PLA2, arachidonic acid, and its metabolites are involved in inflammatory responses, their role after induction of experimental esophagitis was examined. Experiments were performed in esophageal smooth muscle cells (ESO) isolated by enzymatic digestion from the circular layer of normal and esophagitis animals. Content of peptidoleukotrienes (leukotriene [LT] C4, LTD4, and LTE4) was measured in esophageal circular muscle tissue. The cytosolic PLA2 antagonist trifluoromethyl ketone analogue of arachidonic acid inhibited ACh-induced contraction of normal and esophagitis ESO. Inhibition by secreted PLA2 antagonists AM5 and MJ33 was significantly greater in esophagitis ESO. The lipoxygenase inhibitor nordihydro-guaiaretic acid and the LTD4 antagonist ICI 198,615 inhibited ACh-induced contraction of esophagitis but not of normal ESO. Secreted PLA2 and LTD4 contracted normal ESO more than esophagitis ESO. However, in esophagitis, ESO contraction was increased by threshold diacylglycerol concentration. Resting levels of LTs were greater in esophagitis than in normal circular esophageal muscle and increased in response to ACh in esophagitis but not in normal esophageal muscle. Esophagitis shifts the signal transduction pathway activated by ACh. Esophagitis increased the contribution of secreted PLA2 and of LTs to ACh-induced contraction.

Different Protein Kinase C Isozymes Mediate Lower Esophageal Sphincter Tone and Phasic Contraction of Esophageal Circular Smooth Muscle

Different Protein Kinase C Isozymes Mediate Lower Esophageal Sphincter Tone and Phasic Contraction of Esophageal Circular Smooth Muscle

Postprandial motor activity and its relationship to transit in the canine ileum

The purpose of this study was to elucidate the mechanism of reduced intestinal transit rate in the ileum as compared with the jejunum. Twenty-one dogs were each instrumented with 12 strain gauge transducers, 2 collection cannulas, and an infusion catheter defining a 100 cm study in the midjejunum (n = 11) and midileum (n = 10). Postprandial motor activity and intestinal transit were measured 1 hour after ingestion of a 650 kcal solid meal. Contractile activity was analyzed by means of computer programs that determine frequency, amplitude, and propagation behavior of circular smooth muscle contractions. Postprandial ileal contractions occurred with greater frequency (13.7 +/- 2.5 versus 11.5 +/- 0.4; p = 0.04) and displayed a higher incidence of propagation (61% +/- 2% versus 44% +/- 3%; p = 0.0001) than jejunal contractions, but traveled at significantly slower rates (1.0 +/- 0.7 cm/sec vs 3.7 +/- 0.9 cm/sec; p = 0.0001). The net result was significantly slower transit in the ileum compared with the jejunum (4.7 +/- 0.7 cm/min versus 13.1 +/- 1.5 cm/min; p = 0.0006). Within each region, transit correlated with parameters of propagating contractions. Stepwise regression of the combined data revealed that contraction velocity was the most important variable determining intestinal transit rate (r = 0.64; p < 0.001). Contrary to previous thinking, postprandial ileal contractions display a high degree of temporal and spatial organization. Slow ileal transit is mainly due to reduced propagation velocity, which is intrinsic to the circular smooth muscle.

Descending peristaltic reflex in the opossum esophagus

To test the hypothesis that the distension-induced esophageal peristaltic reflex involves a polysynaptic pathway, a triple-chamber organ bath was used to chemically isolate different regions of the opossum smooth muscle esophagus while leaving in continuity the intramural neuromuscular apparatus. Balloon distension in the oral chamber evoked membrane hyperpolarization in the aboral chamber, which was followed, on balloon deflation, by depolarization, spike burst, and circular smooth muscle contraction. This reflex was abolished by adding tetrodotoxin to any of the chambers. Addition of Ca(2+)-free 20 mM Mg2+ Krebs solution (to block synaptic transmission) to the intermediate chamber did not affect the descending peristaltic reflex. However, Ca(2+)-free 20 mM Mg2+ Krebs solution markedly attenuated the reflex when placed in either the oral or the aboral chamber. Furthermore, the nitric oxide synthase inhibitor N omega-nitro-L-arginine methyl ester, when placed in the aboral chamber, abolished the reflex. Other putative neurotransmitter antagonists were without effect when placed in any chamber. These studies suggest that the intramural neural pathway that mediates the descending peristaltic reflex in the opossum esophagus is not polysynaptic, but rather involves long descending neurons that depend on nitric oxide as a final mediator.

Leukotrienes in ACH-induced contraction of esophageal circular smooth muscle in experimental esophagitis: [formula omitted], U.D. Sohn, V. Mangannan, H. Rich, J. Behar, and P. Biancani. Rhode Island Hospital and Brown University, Providence RI

Leukotrienes in ACH-induced contraction of esophageal circular smooth muscle in experimental esophagitis: [formula omitted], U.D. Sohn, V. Mangannan, H. Rich, J. Behar, and P. Biancani. Rhode Island Hospital and Brown University, Providence RI

Progressive changes in cholinergic and tachykinin-mediated circular smooth muscle contractions during TNBS-induced colitis in rats: [formula omitted], J. Goldhill, V.M. Piñero-Carrero, and T. Shea-Donohue. Depts. of Phy., Peds., & Med., USUHS, Bethesda, MD

Progressive changes in cholinergic and tachykinin-mediated circular smooth muscle contractions during TNBS-induced colitis in rats: [formula omitted], J. Goldhill, V.M. Piñero-Carrero, and T. Shea-Donohue. Depts. of Phy., Peds., & Med., USUHS, Bethesda, MD

Relaxatory Effect of Prostaglandin E2 on Circular Smooth Muscle Isolated from the Equine Oviductal Isthmus1

Our objectives were to test whether or not prostaglandin E2 (PGE2) relaxes equine isthmic circular smooth muscle in vitro, and to determine the effects of surgery, time postexcision, and source of tissue on isthmic responsiveness to PGE2. Oviducts were surgically collected after general anesthesia from four research herd mares on Day 5 postovulation, after stunning and exsanguination from four research herd mares on Day 5, and from seven slaughterhouse mares during diestrus. Circular smooth muscle contractions were measured in vitro with an isometric force transducer. Tissues were precontracted with 2 μM histamine and then were treated with PGE2 (0, 0.1, 1, 10, 100, or 1000 nM). Histamine treatment significantly (p < 0.001) increased tissue tension (500.0 ± 0.0 to 818.9 ± 47.9 mg) and contractile frequency (13.3 ± 0.4 to 29.8 ± 1.6 contractions/min), and decreased contractile amplitude (69.7 ± 11.9 to 23.7 ± 4.2 mg). PGE2 treatment significantly (p < 0.001) decreased tissue tension and contractile frequency and significantly (p < 0.001) increased contractile amplitude of histamine-precontracted tissue in a dose-dependent manner. Tissue obtained from mares anesthetized on Day 5 was significantly (p < 0.05) less responsive to PGE2 than tissue obtained from mares killed on Day 5. Responsiveness to PGE2 of tissue from mares killed on Day 5 did not decrease significantly after 12 h of storage at 4°C. These results support the hypothesis that PGE2 relaxes isthmic circular smooth muscle, and indicate that anesthetic agents reduce isthmic responsiveness to PGE2.

GABA receptors in the dorsal motor nucleus of the vagus influence feline lower esophageal sphincter and gastric function

Gamma-aminobutyric acid (GABA) antagonist (bicuculline methiodide, BIC; picrotoxin, PIC) or agonist (muscimol, MUS) microinjections were made into the dorsal motor nucleus of the vagus nerve (DMV), and effects on lower esophageal sphincter pressure (LESP), gastric motility, and gastric acid secretion were determined in chloralose-anesthetized cats. Right or left DMV sites were microinjected with BIC, PIC, MUS, or isotonic tonic saline (140 nl) through a glass micropipette having a tip diameter of 15-21 microns. Esophageal body, LESP, and gastric fundic pressures were measured manometrically. Circular smooth muscle contractions of the antrum and pylorus were recorded with strain-gauge force transducers. Gastric acid secretion was measured every 15 min through a gastric cannula and titrated to pH 7.0. DMV microinjection sites were verified histologically. Direct BIC microinjections (0.275 or 0.550 nmol) into the DMV primarily produced a decrease in LESP (71% of all sites tested), with mean LESP changing from 23.2 +/- 1.7 mmHg to 3.7 +/- 0.7 mmHg (p < 0.01). Tonic LESP increases and phasic LESP contractile activity occurred less frequently. BIC-induced LESP responses were abolished by vagotomy or by microinjections of MUS (0.5 to 10 nmol) into the DMV. Direct PIC microinjection (0.232 nmol) into the DMV produced a pattern of responses similar to those observed with BIC (which were also abolished by vagotomy or by MUS microinjections into the DMV). The antrum and pylorus were also responsive to DMV microinjections of both GABA antagonists. Microinjections of BIC or PIC into the DMV produced increases in gastric circular muscle activity that occurred less frequently than LESP effects, but also were eliminated by vagotomy. The high (0.550 nmol) dose of BIC increased gastric motility significantly more often than the low dose of BIC (p < 0.05). In addition, BIC (0.550 nmol) microinjections into the DMV increased gastric secretory volume (from 0.6 +/- 0.2 to 6.0 +/- 2.5 ml/15 min; p < 0.01) and total titratible acid (from 34.4 +/- 8.9 to 86.0 +/- 19.1 mEq/15 min; p < 0.01), and decreased gastric pH (from 4.63 +/- 0.44 to 3.50 +/- 0.49; p < 0.05). Vagotomy also eliminated the gastric secretory effects of DMV BIC. Direct microinjections of MUS into the DMV also blocked BIC- or PIC-induced changes in gastric motility and/or gastric acid secretion. Isotonic saline microinjected into the DMV did not increase basal or decrease stimulated gastric esophageal motility or gastric secretion. These data indicate that LESP, gastric motility, and gastric secretion are influenced by a tonic DMV inhibition mediated by GABAA receptor stimulation of the DMV. Because disinhibition of these receptors clearly activates the upper gut, future work should focus on identifying the nuclei providing this synaptic input to the DMV that might be involved in the functional regulation of upper gut motor and secretory function.

Evidence for maturational changes in phosphlipase activity and sources of calcium utilized for contraction in the feline antrum

EVIDENCE FOR MATURATIONAL CHANGES IN PHOSPHLIPASE ACTIVITY AND SOURCES OF CALCIUM UTILIZED FOR CONTRACTION IN THE FELINE ANTRUM. C. Hillemeier, D. Deutsch, and K.N. Bitar. Department of Pediatrics, Umv. of Michigan, Ann Arbor, MI 48109. Unlike isolated circular smooth muscle cells from the feline adult gastric annum, feline newborn antral cells are unable to utilize intracellular calcium stores to contract in response to myogenic agonists. Cholecystokinin octapeptide (CCK-OP) induces dose dependent contraction in both newborn and adult antral cells. The contractile response to CCK is relatively transient in adult cells while sustained in kitten antral cells. U73122 blocks the intial contraction in adult antral cells but not the sustained contraction seen in kitten antral cells. In adult antral tissue CCK-OP (lo-7M) causes an early transient increase in the production of both 1, 2 Diaglylcerol (DAG), while in the newborn antrum CCK-OP caused a sustained increase in DAG production for up to four minutes. Measurement of 1, 4, 5 inositol trisphosphate (IP3), which is known to release intracellular calcium stores, showed an early increase in both adult and kitten antral cells. We have measured changes in calcium concentrations in isolated antral smooth muscle cells by digital imaging spectrofluoroscopy in cells loaded with Fura 2. Newborn antral cells show a rise in intracellular calcium in response to agonist induced stimulation. Incubation of the newborn annal cells with the Ca ATPase inhibitor thapsigargin, which leads to the emptying of intracellular calcium stores, does not result in an increase in intracellular calcium levels, suggesting that intracellular calcium stores are not readily available. In summary the contractile response to CCK-OP in the newborn is associated with an initial rise in IP3 production and sustained DAG production, while contraction in adult antral cells is associated with transient increase in both IP3 and DAG. Compared to the adult, agonist induced contraction in the newborn kitten antrum may be associated with both quantitative and qualitative differences in phospholipase activity. This may be necessitated because of a relative inaccessibilty of intracellular calcium stores in the newborn kitten annum. The maturational changes which are seen in the intracellular pathways associated with antral circular smooth muscle contraction may be reflected in chantin~functional capabilities of this region.

Contractions mediated by alpha 1-adrenoceptors and P2-purinoceptors in a cat colon circular muscle.

1. The postjunctional excitatory and inhibitory effects of adrenoceptor and purinoceptor agonists and antagonists were studied in circular smooth muscle strips of cat colon. 2. In the presence of tetrodotoxin (0.5 microM), noradrenaline caused contraction or relaxation of circular smooth muscle at resting tension or with raised tone, respectively. The noradrenaline-evoked contractions were potentiated and the noradrenaline-evoked relaxations were antagonized by propranolol (1 microM), suggesting beta-adrenoceptor involvement. 3. At resting tension, noradrenaline, adrenaline and the selective alpha 1-adrenoceptor agonist, phenylephrine, caused concentration-dependent contractile responses, with EC50 values of 1.8 +/- 0.2 microM, 1.9 +/- 0.4 microM and 4.3 +/- 1.7 microM, respectively. The EC50 values and the amplitude of maximal responses were not significantly different from one another. Clonidine (0.1-500 microM), a selective alpha 2-adrenoceptor agonist, was not effective. 4. Prazosin (0.1-9 microM), competitively antagonized the contractile effects of noradrenaline with an estimated pA2 value of 6.93 and a slope of 1.07 +/- 0.03. The Kb values, estimated from a single shift (0.1 microM prazosin) of the concentration-response curves to noradrenaline, adrenaline and phenylephrine were 92.8 +/- 9.3 nM, 108.7 +/- 6.4 nM and 18.4 +/- 3.1 nM, respectively. 5. At resting tension, adenosine 5' triphosphate (ATP, 5-1000 microM), alpha,beta-methylene adenosine 5'-triphosphate (alpha,beta-MeATP, 0.05-50 microM), beta,gamma-methylene adenosine 5'-triphosphate (beta,gamma-MeATP, 0.5-100 microM), and 2-methylthioadenosine 5'-triphosphate (2-MeSATP, 1-500 microM) caused concentration-dependent contractions with EC50 values of 60.5 +/- 15.9 microM, 0.7 +/- 0.1 microM, 7.6 +/- 0.1 microM and 25.3 +/- 12.8 microM, respectively. The maximal responses to alpha, beta-MeATP and beta,gamma-MeATP were greater than maximal responses to 2-MeSATP and ATP.6. Suramin (50-500 microM), competitively antagonized the contractile responses of alpha,beta-MeATP with an estimated pA2 value of 4.92 and a slope of 1.08 +/- 0.04. The Kb values, estimated from a single shift(1I00 microM suramin) of the concentration-response curves to ATP, alpha,beta-MeATP, beta,gamma-MeATP and 2MeSATPwere 52.3 +/- 20.2 microM, 25.2 +/- 4.5 microM, 21.7 +/- 11.0 microM and 11.6 +/- 2.7 microM, respectively.7. At resting tension, reactive blue 2 (100 microM), a selective antagonist of the P2Y-purinoceptor, and 8-(p-sulphophenyl)-theophylline (8-SPT) (1 microM), a selective antagonist of the PI-purinoceptor, did not antagonize the contractile responses to alpha,beta-MeATP (0.5-5 microM). Contractile responses to ATP(50-500 microM) were not altered by 8-SPT (I microM) but were potentiated by reactive blue 2 (100 microM).8. With raised tone, ATP and 2-MeSATP caused a relaxant effect. This effect of ATP was not altered by either tetrodotoxin (TTX) (0.5 microM) or suramin (100 microM), but was antagonized by reactive blue 2(100 microM) and 8-SPT (1 microM), suggesting that inhibitory P2y- and P1-purinoceptors are involved. In contrast, alpha,beta-MeATP and Beta,gamma-MeATP caused only contractions. This contractile effect of alpha,beta-MeATPwas resistant to TTX (0.5 microM) and antagonized by suramin (100 microM).9. In summary, cat colon circular muscle contains postjunctional alpha 1-adrenoceptors and P2X purinoceptors which mediate contractions and P2y- and PI-purinoceptors which mediate relaxation.Postjunctional alpha2-adrenoceptors appear not to be present.

Acquired megacolon is associated with alteration of vasoactive intestinal peptide levels and acetylcholinesterase activity

Based upon previous morphologic studies, we hypothesized that the development of acquired megacolon was associated with abnormalities of enteric neurotransmitter concentrations and enzymatic activities. Specimens were obtained at surgery from patients with normal descending-sigmoid colon (n = 13) and patients with sigmoid megacolon (n = 6; defined by radiologic measurement). Radioimmunoassays were used to measure the non-adrenergic, non-cholinergic inhibitory neuropeptide, vasoactive intestinal peptide, and the non-adrenergic, non-cholinergic excitatory neuropeptide, substance P, while spectrophotometric assays were used to quantitate acetylcholinesterase activity and choline acetyltransferase activity. There were significantly decreased concentrations of vasoactive intestinal peptide and decreased acetylcholinesterase activity in muscularis externa from patients with acquired megacolon. In megacolon, vasoactive intestinal peptide-containing nerve fibers appeared to be diminished in circular and longitudinal smooth muscle, and immunostaining of nerve cell bodies in the plexus submucosus externus appeared diminished. These results suggest the hypothesis that production of vasoactive intestinal peptide is altered allowing secondary colonic hypertrophy to develop from prolonged cholinergic nerve-mediated contractions of circular smooth muscle. As a corollary to this hypothesis, colonic dilatation might result from prolonged contraction of longitudinal smooth muscle.

Mediation of anaphylaxis-induced jejunal circular smooth muscle contraction in rats

Altered intestinal motility and diarrhea are features of food protein-induced intestinal anaphylaxis in the conscious rat. These experiments were performed to determine the mediator(s) responsible for jejunal circular smooth muscle contraction during this response. Hooded-Lister rats were sensitized by intraperitoneal injection of 10-micrograms egg albumin, and controls were sham-sensitized with saline. Fourteen days later the contractility of the circular muscle in jejunal segments (mucosa intact) was examined in standard tissue baths in response to antigen (Ag) or other agents. While control and sensitized tissues contracted in similar fashion in response to stretch, bethanechol, histamine, or 5-hydroxytryptamine (5HT), Ag contracted only the segments of sensitized animals. The contractile response was: (1) specific to the sensitizing Ag, as bovine serum albumin did not induce contraction and (2) could be passively transferred with serum containing specific immunoglobulin E antibody (IgE-Ab). Concanavalin A, which degranulates both mucosal and connective tissue-type mast cells, and compound 48/80, which degranulates only connective tissue-type mast cells produced contractile responses. Ag-induced contraction was significantly inhibited by the mucosal and connective tissue-type mast cell stabilizer doxantrazole, but not the connective tissue mast cell stabilizer disodium cromoglycate. Diphenhydramine and cimetidine together significantly inhibited histamine-induced contraction, but failed to effect the Ag-induced contraction in sensitized tissues. While the contractile response to 5HT was reduced in the presence of methysergide (5HT1-receptor antagonist), cinanserin (5HT2-receptor antagonist), and ICS 205-930 (5HT3-receptor antagonist), only cinanserin significantly inhibited the contractile response to Ag. Indomethacin significantly inhibited Ag-induced contraction. Ag-induced contraction was resistant to atropine and tetrodotoxin.(ABSTRACT TRUNCATED AT 250 WORDS)