Gastrointestinal Muscle Research Articles

Actividad antimicrobiana, antiespasmódica y acción sobre hormonas sexuales en ratas hembras, de los componentes de Tagetes Pusilla (Anisillo)

En la presente investigación se detectó e identificó los tipos de metabolitos secundarios predominantes en el extracto etanólico de las hojas de Tagetes pusilla "anisillo" "anís serrano", que fueron compuestos fenólicos, flavonoides y alcaloides. Se determinó la actividad antibacteriana comparativa entre el aceite esencial y el extracto etanólico de anisillo, encontrándose que ambos mostraron una actividad antibacteriana significativa. Comprobándose la actividad de los metabolitos secundarios presentes en el extracto etanólico de hojas de Tagetes pusilla sobre el músculo liso gastrointestinal, mediante los métodos de Chidume et al 2002 con modificaciones; y el de Akah et al 1998; así como la determinación del nivel de prolactina en ratas según Di Carlo et al 2005 con modificadones, encontrándose que el extracto etanólico de Tagetes pusilla en las condiciones experimentales aumentó los niveles de prolactina, estrógenos, progesterona, LH, FSH en ratas normales; habiéndose evidenciado relajación del duodeno aislado de rata, y se observó un incremento de la actividad del ileon de rata, que probablemente se deba a la presencia de alcaloides.

A Full Term Infant with Cyanotic Episodes

A Full Term Infant with Cyanotic Episodes

The α2-adrenergic receptor antagonist yohimbine improves endotoxin-induced inhibition of gastrointestinal motility in mice

Sepsis inhibits gastrointestinal motility. Although the exact mechanism of this is unclear, lipopolysaccharide is known to activate macrophages in the gastrointestinal wall, which upregulate their expression of inducible nitric oxide synthase (iNOS). This leads to an increased production of nitric oxide, which relaxes the gastrointestinal muscles. We studied endotoxaemic mice to determine whether yohimbine improved delayed gastric emptying and gastrointestinal transit. Male Balb/c mice (n = 49) were randomly allocated to two groups, and either yohimbine 25 microg or saline was injected s.c. Four hours later, mice in each group were further randomly allocated to two groups, and either lipopolysaccharide 100 microg or saline was injected intraperitoneally. Eight hours later, liquid containing fluorescent microbeads was infused into the stomach, and 30 min later, gastric emptying and gastrointestinal transit were measured using flow cytometry. We also studied whether yohimbine given after injection of lipopolysaccharide was effective (n = 22). In another group of mice (n = 32), iNOS in the gastrointestinal tract was measured using western blotting. Lipopolysaccharide significantly inhibited gastric emptying and gastrointestinal transit. Yohimbine, given before or after lipopolysaccharide, significantly attenuated the inhibitory effects of lipopolysaccharide. Lipopolysaccharide increased the expression of iNOS in the small intestine and yohimbine suppressed the effects of lipopolysaccharide. In endotoxaemic mice, yohimbine improved delayed gastric emptying and gastrointestinal transit, possibly by downregulating lipopolysaccharide-induced increased expression of iNOS.

Interstitial cells of Cajal in the gastrointestinal musculature of W mutant mice

Interstitial cells of Cajal (ICC) are important regulatory cells generating electrical rhythmicity and transducing neural signals in the gastrointestinal musculature. ICC express the proto-oncogene c-kit, a receptor tyrosine kinase, and can be examined morphologically using the c-Kit antibody. The c-kit gene is allelic with the murine white-spotting locus W, and the c-kit mutation (W mutation) affects various aspects of hematopoietic cells, germ cells, melanocytes, mast cells, and ICC. Heterozygous W/W( v) mutant mice lack a specific type of ICC and have been used to reveal its function. To search for a new model that lacks a specific type of ICC, we examined homozygous W( v)/W( v) black-eyed-white mice that are viable with anemia. Results showed the principal patterns of ICC deficiency were the same between the W/W( v) and W( v)/W( v) mutants. In the stomach of both mice, intramuscular ICC (ICC-IM) were missing and myenteric ICC (ICC-MY) were reduced in number. In the small intestine, the number of ICC-MY was severely reduced in spite of a normal distribution of deep muscular plexus ICC (ICC-DMP). The cecum also exhibited fewer reduced. ICC-IM in the colon were almost entirely missing, whereas ICC-MY were reduced only in the distal colon. In the small intestine and colon, the number of remaining ICC-MY in W( v)/W( v) mice was greater than that in W/W( v) mice. The enteric nervous system of the two mutant mice showed normal characteristics. From these findings, we conclude that W( v)/W( v) mice represent a new genotype that lacks a part of the ICC in its gastrointestinal musculature.

Messenger RNA Levels and Binding Sites of Muscarinic Acetylcholine Receptors in Gastrointestinal Muscle Layers from Healthy Dairy Cows

Acetylcholine interacts with muscarinic receptors (M) to mediate gastrointestinal (GI) smooth muscle contractions. We have compared mRNA levels and binding sites of M1to M5 in muscle tissues from fundus abomasi, pylorus, ileum, cecum, proximal loop of the ascending colon (PLAC), and external loop of the spiral colon (ELSC) of healthy dairy cows. The mRNA levels were measured by quantitative RT-PCR. The inhibition of [3H]-QNB (1-quinuclidinyl-[phenyl-4-3H]-benzilate) binding by M antagonists [atropine (M1 − 5), pirenzepine (M1), methoctramine (M2), 4-DAMP (M3), and tropicamide (M4)] was used to identify receptors at the functional level. Maximal binding (Bmax) was determined through saturation binding with atropine as a competitor. The mRNA levels of M1, M2, M3, and M5 represented 0.2, 48, 50, and 1.8%, respectively, of the total M population, whereas mRNA of M4 was undetectable. The mRNA levels of M2 and of M3 in the ileum were lower (P < 0.05) than in other GI locations, which were similar among each other. Atropine, pirenzepine, methoctramine, and 4-DAMP inhibited [3H]-QNB binding according to an either low- or high-affinity receptor pattern, whereas tropicamide had no effect on [3H]-QNB binding. The [3H]-QNB binding was dose-dependent and saturable. Bmax in fundus, pylorus, and PLAC was lower (P < 0.05) than in the ELSC, and in the pylorus lower (P < 0.05) than in the ileum. Bmax and mRNA levels were negatively correlated (r = -0.3; P < 0.05). In conclusion, densities of M are different among GI locations, suggesting variable importance of M for digestive functions along the GI tract.

Kit mutants and gastrointestinal physiology

There has been considerable speculation about the function of interstitial cells of Cajal (ICC) since their discovery more than 100 years ago. It has been difficult to study these cells under native conditions, but great insights about the function of ICC have come from studies of genetic models with loss-of function mutations in the Kit signalling pathway. First it was discovered that signalling via Kit (a receptor tyrosine kinase) was vital for the development and maintenance of the ICC phenotype in gastrointestinal (GI) muscles. In compound heterozygotes (W/W(V) and Sl/Sl(d) animals), where there are partial loss-of-function mutations in Kit receptors or Kit ligand (stem cell factor), ICC failed to develop in various regions of the GI tract, but no major changes in the smooth muscle layers or enteric nervous system occurred in the absence of these cells. Animals with these mutations provided an unprecedented opportunity to understand the role of ICC in GI motor function, and it is now clear from these studies that ICC serve as: (i) pacemaker cells, generating the spontaneous electrical rhythms of the gut known as slow waves; (ii) a propagation pathway for slow waves so that large areas of the musculature can be entrained to a dominant pacemaker frequency; (iii) mediators of excitatory cholinergic and inhibitory nitrergic neural inputs from the enteric nervous system, and (iv) stretch receptors that modulate membrane potential and electrical slow wave frequency. This review describes the use of genetic models to understand the important physiological role of ICC in the GI tract.

5-Hydroxytryptamine-4 receptor messenger ribonucleic acid levels and densities in gastrointestinal muscle layers from healthy dairy cows1

Serotonin (5-hydroxytryptamine, 5-HT) is involved in gastrointestinal tract (GIT) motor functions through binding to specific receptors located in the GIT walls. The objectives of the current study were to compare mRNA levels and binding sites of 5-HT(4) receptors (5-HTR(4)) in smooth muscle layers from the fundus abomasi, pylorus, ileum, cecum, proximal loop of the ascending colon (PLAC), and external loop of the spiral colon (ELSC) of healthy dairy cows, and to verify whether mRNA and protein expression were correlated. Smooth muscle samples were prepared by scraping the mucosa and submucosa from full-thickness intestinal wall samples. The mRNA levels of 5-HTR(4) were measured by real-time PCR and expressed relative to those of the housekeeping gene glyceraldehyde phosphate dehydrogenase. Binding studies were performed using the 5-HTR(4) antagonist [(3)H]GR113808. The mRNA levels of 5-HTR(4) were affected (P < 0.05) by location along the GIT. The mRNA levels of 5-HTR(4) in the ELSC and the ileum were greater than in the PLAC (P = 0.05 and P = 0.07, respectively) but similar to those of all other locations. The competitive binding of [(3)H]GR113808 to suspended membranes from the fundus abomasi, pylorus, cecum, and ELSC was best fit by a 2-site receptor model, whereas it was best fit by a 1-site receptor model in the ileum and PLAC. The mRNA levels and numbers of 5-HTR(4) were not correlated (r = 0.14; P = 0.71). In conclusion, mRNA and binding sites for 5-HTR(4) are present in the smooth muscle layer of the entire GIT of dairy cows and may play a role with respect to motility. The effects of activation of this receptor subtype may be different among GIT locations due to differences in the amount of high- relative to low-affinity binding sites.

Pacemaker phase shift in the absence of neural activity in guinea‐pig stomach: a microelectrode array study

Gastrointestinal (GI) motility is well organized. GI muscles act as a functional syncytium to achieve physiological functions under the control of neurones and pacemaker cells, which generate basal spontaneous pacemaker electrical activity. To date, it is unclear how spontaneous electrical activities are coupled, especially within a micrometre range. Here, using a microelectrode array, we show a spatio-temporal analysis of GI spontaneous electrical activity. The muscle preparations were isolated from guinea-pig stomach, and fixed in a chamber with an array of 8 x 8 planar multielectrodes (with 300 microm in interpolar distance). The electrical activities (field potentials) were simultaneously recorded through a multichannel amplifier system after high-pass filtering at 0.1 Hz. Dihydropyridine Ca(2+) channel antagonists are known to differentiate the electrical pacemaker activity of interstitial cells of Cajal (ICCs) by suppressing smooth muscle activity. In the presence of nifedipine, we observed spontaneous electrical activities that were well synchronized over the array area, but had a clear phase shift depending on the distance. The additional application of tetrodotoxin (TTX) had little effect on the properties of the electrical activity. Furthermore, by constructing field potential images, we visualized the synchronization of pacemaker electrical activities resolving phase shifts that were measurable over several hundred micrometres. The results imply a phase modulation mechanism other than neural activity, and we postulate that this mechanism enables smooth GI motility. In addition, some preparations clearly showed plasticity of the pacemaker phase shift.

Motor disturbances participate in the pathogenesis of eosinophilic oesophagitis, beyond the fibrous remodelling of the oesophagus

Motor disturbances participate in the pathogenesis of eosinophilic oesophagitis, beyond the fibrous remodelling of the oesophagus

G.P.10 08 Lower urinary tract symptoms in patients with Duchenne muscular dystrophy

Duchenne muscular dystrophy (DMD) is a progressive disease that mainly affects striated muscles. However, involvement of gastrointestinal muscles (smooth muscles) and bladder dysfunction have also been reported. Lower urinary tract symptoms (LUTS) are frequently reported by boys with DMD at the outpatient rehabilitation clinic, though the prevalence of LUTS and the effect of LUTS on social activities in the DMD population are unknown. Therefore, we conducted a questionnaire-based survey on the prevalence and social effects of LUTS in the Dutch DMD population.

Evidence-Based Perianesthesia Care: Accelerated Postoperative Recovery Programs

Prolonged stress response after surgery can cause numerous adverse effects, including gastrointestinal dysfunction, muscle wasting, impaired cognition, and cardiopulmonary, infectious, and thromboembolic complications. These events can delay hospital discharge, extend convalescence, and negatively impact long-term prognosis. Recent advances in perioperative management practices have allowed better control of the stress response and improved outcomes for patients undergoing surgery. At the center of the current focus on improved outcomes are evidence-based fast-track surgical techniques and what is commonly referred to as "accelerated postoperative recovery programs." These programs require a multidisciplinary, coordinated effort, and nurses are essential to their successful implementation.

Biliverdin protects against polymicrobial sepsis by modulating inflammatory mediators

Highly inducible heme oxygenase (HO)-1 is protective against acute and chronic inflammation. HO-1 generates carbon monoxide (CO), ferrous iron, and biliverdin. The aim of this study was to investigate the protective effects of biliverdin against sepsis-induced inflammation and intestinal dysmotility. Cecal ligation and puncture (CLP) was performed on Sprague-Dawley rats under isoflurane anesthesia with and without intraperitoneal biliverdin injections, which were done before, at the time of CLP, and after CLP. In vivo gastrointestinal transit was carried out with fluorescein-labeled dextran. Jejunal circular muscle contractility was quantified in vitro using organ bath-generated bethanechol dose-response curves. Neutrophilic infiltration into the muscularis externa was quantified. The jejunal muscularis was studied for cytokine mRNA expressions [interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1, inducible nitric oxide synthase, cyclooxygenase-2, biliverdin, IL-10, and HO-1] using real-time RT-PCR. Biliverdin treatment prevented the sepsis-induced suppression of gastrointestinal muscle contractility in vivo and in vitro and significantly decreased neutrophilic infiltration into the jejunal muscularis. Inflammatory mRNA expressions for small bowel IL-6 and MCP-1 were significantly reduced after biliverdin treatment in CLP-induced septic animals compared with untreated septic animals. The anti-inflammatory mediator expression of small bowel IL-10 was significantly augmented after CLP at 3 h compared with untreated septic animals. These findings demonstrate that biliverdin attenuates sepsis-induced morbidity to the intestine by selectively modulating the inflammatory cascade and its subsequent sequelae on intestinal muscularis function.

Interstitial Cells of Cajal Are Involved in Neurotransmission in the Gastrointestinal Tract

Interstitial cells of Cajal (ICC) are important cells which coordinate gastrointestinal motility. ICC express Kit receptor tyrosine kinase, and Kit immunohistochemistry reveals ICC morphology and distribution in the gastrointestinal musculature. ICC show a highly branched morphology and form unique networks. Myenteric ICC (ICC-MY) are located at the layer of the myenteric plexus and serve as electrical pacemakers. Intramuscular ICC (ICC-IM) and ICC in the deep muscular plexus (ICC-DMP) are distributed within the muscular layers, and are densely innervated by excitatory and inhibitory enteric motor neurons and in close contact with nerve terminals. Recent studies combined with morphological and functional techniques directly revealed that ICC-IM and ICC-DMP are mediators of enteric motor neurotransmission. These types of ICC express several receptors for neurotransmitters such as acetylcholine and substance P and show responses to excitatory nerve stimulations. ICC also express receptive mechanisms for nitric oxide, which is an inhibitory neurotransmitter in the gastrointestinal tract. They can respond to nitrergic nerve stimulation by cyclic GMP production. Kit mutant mice lack ICC-IM and show attenuated postsynaptic responses after intrinsic nerve stimulation. These findings indicate the importance for ICC in neurotransmission in the gastrointestinal tract.

Muscarinic M 2 acetylcholine receptor distribution in the guinea-pig gastrointestinal tract

In the enteric nervous system, acetylcholine is the most common neurotransmitter to induce gastrointestinal smooth muscle contractions. Cholinergic signaling is mediated by muscarinic acetylcholine receptors on the surface of smooth muscle cells. Five different muscarinic receptor subtypes (M 1–M 5) have been identified and characterized, all of which belong to the superfamily of the G-protein-coupled receptor. The muscarinic M 2 acetylcholine receptor is the major muscarinic receptor subtype expressed by smooth muscle tissues in the gastrointestinal tract, where it is coexpressed with a smaller population of M 3 receptor. In this study, we examined the immunohistochemical distribution of the M 2 receptor using a specific antibody in the guinea-pig gastrointestinal tract. M 2 receptor-like immunoreactivity was mainly observed as associated with smooth muscle cells in the gastrointestinal tract. M 2 receptor-like immunoreactivity in smooth muscle cells was distributed throughout the cell membrane associated with caveolae. In the proximal colon, M 2 receptor-like immunoreactivity in the smooth muscle cells was weak. In the small intestine, interstitial cells of Cajal that possessed neurokinin 1 receptor-like immunoreactivity had intense M 2 receptor-like immunoreactivity. In the proximal colon, intramuscular and myenteric interstitial cells of Cajal exhibited M 2 receptor-like immunoreactivity. These findings indicate that, in the gastrointestinal musculature, M 2 receptors are distributed both in the smooth muscle cells and interstitial cells of Cajal, suggesting that the M 2 receptor elicits smooth muscle cell contraction and the interstitial cells of Cajal are the sites of innervation by enteric cholinergic neurons.

Activating of ATP-Dependent K+ Channels Comprised of Kir 6.2 and SUR 2B by PGE2 Through EP2 Receptor in Cultured Interstitial Cells of Cajal from Murine Small Intestine

The interstitial cells of Cajal (ICC) are pacemaker cells in gastrointestinal tract and generate an electrical rhythm in gastrointestinal muscles. We investigated the possibility that PGE₂ might affect the electrical properties of cultured ICC by activating ATPdependent K⁺ channels and, the EP receptor subtypes and the subunits of ATP-dependent K⁺ channels involved in these activities were identified. In addition, the regulation of intracellular Ca²⁺ ([Ca²⁺]i) mobilization may be involved the action of PGE₂ on ICC. Treatments of ICC with PGE₂ inhibited electrical pacemaker activities in the same manner as pinacidil, an ATPdependent K⁺ channel opener and PGE₂ had only a dose-dependent effect. Using RT-PCR technique, we found that ATP-dependent K⁺ channels exist in ICC and that these are composed of K_ir 6.2 and SUR 2B subunits. To characterize the specific membrane EP receptor subtypes in ICC, EP receptor agonists and RT-PCR were used: Butaprost (an EP₂ receptor agonist) showed the actions on pacemaker currents in the same manner as PGE₂. However sulprostone (a mixed EP₁ and EP₃ agonist) had no effects. In addition, RT-PCR results indicated the presence of the EP₂ receptor in ICC. To investigate cAMP involvement in the effects of PGE₂ on ICCs, SQ-22536 (an inhibitor of adenylate cyclase) and cAMP assays were used. SQ-22536 did not affect the effect of PGE₂ on pacemaker currents, and PGE₂ did not stimulate cAMP production. Also, we found PGE₂ inhibited the spontaneous [Ca²⁺]i oscillations in cultured ICC. These observations indicate that PGE₂ alters pacemaker currents by activating the ATP-dependent K⁺ channels comprised of K_ir 6.2-SUR 2B in ICC and this action of PGE₂ are through EP₂ receptor subtype and also the activation of ATP-dependent K⁺ channels involves intracellular Ca²⁺ mobilization.

New perspectives of studying gastrointestinal muscle function

The motor function of the gastrointestinal tract has primarily been studied using manometry and radiography, though more indirect tests have also been applied. Manometry and radiography do not provide detailed information about the muscle properties as can be assessed from studies of muscle properties in muscle strips in vitro. In recent years a technique based on impedance planimetric measurement of pressure-cross-sectional area relations in a distending bag has proven to provide more detailed information about the muscle function in vivo. This review shows examples of new muscle function analysis such as length-tension diagrams, force-velocity curves and preload-afterload diagrams.

Spatial and temporal mapping of pacemaker activity in interstitial cells of Cajal in mouse ileum in situ

Spontaneous electrical pacemaker activity occurs in tunica muscularis of the gastrointestinal tract and drives phasic contractions. Interstitial cells of Cajal (ICC) are the pacemaker cells that generate and propagate electrical slow waves. We used Ca(2+) imaging to visualize spontaneous rhythmicity in ICC in the myenteric region (ICC-MY) of the murine small intestine. ICC-MY, verified by colabeling with Kit antibody, displayed regular Ca(2+) transients that occurred after electrical slow waves. ICC-MY formed networks, and Ca(2+) transient wave fronts propagated through the ICC-MY networks at approximately 2 mm/s and activated attached longitudinal muscle fibers. Nicardipine blocked Ca(2+) transients in LM but had no visible effect on the transients in ICC-MY. beta-Glycyrrhetinic acid reduced the coherence of propagation, causing single cells to pace independently. Thus, virtually all ICC-MYs are spontaneously active, but normal activity is organized into propagating wave fronts. Inhibitors of dihydropyridine-resistant Ca(2+) entry (Ni(2+) and mibefradil) and elevated external K(+) reduced the coherence and velocity of propagation, eventually blocking all activity. The mitochondrial uncouplers, FCCP, and antimycin and the inositol 1,4,5-trisphosphate receptor-inhibitory drug, 2-aminoethoxydiphenyl borate, abolished rhythmic Ca(2+) transients in ICC-MY. These data show that global Ca(2+) transients in ICC-MYs are a reporter of electrical slow waves in gastrointestinal muscles. Imaging of ICC networks provides a unique multicellular view of pacemaker activity. The activity of ICC-MY is driven by intracellular Ca(2+) handling mechanisms and entrained by voltage-dependent Ca(2+) entry and coupling of cells via gap junctions.

Two‐pore‐domain potassium channels in smooth muscles: new components of myogenic regulation

Gastrointestinal (GI) smooth muscles are influenced by many levels of regulation, including those provided by enteric motor neurones, hormones and paracrine substances. The integrated contractile responses to these regulatory mechanisms depend heavily on the state of excitability of smooth muscle cells. Resting ionic conductances and myogenic responses to agonists and physical parameters, such as stretch, are important in establishing basal excitability. This review discusses the role of 2-pore-domain K+ channels in contributing to background conductances and in mediating responses of GI muscles to enteric inhibitory nerve stimulation and stretch. Murine GI muscles express TREK-1 channels and display a stretch-dependent K+ (SDK) conductance that is also activated by nitric oxide via a cGMP-dependent mechanism. Cloning and expression of mTREK-1 produced an SDK conductance that was activated by cGMP-dependent phosphorylation at serine-351. GI muscle cells also express TASK-1 and TASK-2 channels that are inhibited by lidocaine and external acidification. These conductances appear to provide significant background K+ permeability that contributes to the negative resting potentials of GI muscles.

A pH‐sensitive potassium conductance (TASK) and its function in the murine gastrointestinal tract

The excitability of smooth muscles is regulated, in part, by background K+ conductances that determine resting membrane potential. However, the K+ conductances so far described in gastrointestinal (GI) muscles are not sufficient to explain the negative resting potentials of these cells. Here we describe expression of two-pore K+ channels of the TASK family in murine small and large intestinal muscles. TASK-2, cloned from murine intestinal muscles, resulted in a pH-sensitive, time-dependent, non-inactivating K+ conductance with slow activation kinetics. A similar conductance was found in native intestinal myocytes using whole-cell patch-clamp conditions. The pH-sensitive current was blocked by local anaesthetics. Lidocaine, bupivacaine and acidic pH depolarized circular muscle cells in intact muscles and decreased amplitude and frequency of slow waves. The effects of lidocaine were not blocked by tetraethylammonium chloride, 4-aminopyridine, glibenclamide, apamin or MK-499. However, depolarization by acidic pH was abolished by pre-treatment with lidocaine, suggesting that lidocaine-sensitive K+ channels were responsible for pH-sensitive changes in membrane potential. The kinetics of activation, sensitivity to pH, and pharmacology of the conductance in intestinal myocytes and the expression of TASK-1 and TASK-2 in these cells suggest that the pH-sensitive background conductance is encoded by TASK genes. This conductance appears to contribute significantly to resting potential and may regulate excitability of GI muscles.

Sulfur‐containing amino acids block stretch‐dependent K+ channels and nitrergic responses in the murine colon

1. Efforts to determine the role of stretch-dependent K(+) (SDK) channels in enteric inhibitory neural responses in gastrointestinal muscles are difficult due to a lack of blocking drugs for SDK channels. 2. SDK channels are blocked by sulfur-containing amino acids. These compounds reduced the open probability of SDK channels in on and off-cell patches of murine colonic myocytes. L-Methionine was the most selective and had little or no effect on other known K(+) conductances in colonic myocytes. 3. Application of L-cysteine, L-methionine or DL-homocysteine depolarized intact muscles and enhanced spontaneous contractions. D-Stereoisomers of these amino acids were less effective than L-stereoisomers. 4. Pretreatment of muscles with tetrodotoxin, N(W)-nitro-L-arginine or 1H-[1,2,4] oxadiazolo [4,3-a] quinoxalin-1-one reduced the depolarization responses to these compounds, suggesting that spontaneous neural activity and release of NO tonically activates SDK channels. 5. Nitrergic responses to nerve stimulation were reduced by sulfur-containing amino acids. 6. These data suggest that nitrergic inhibitory junction potentials are mediated, in part, by activation of SDK channels in murine colonic muscles.