Effect of intraluminal distension or ischemic strangulation obstruction of the equine jejunum on jejunal motilin receptors and binding of erythromycin lactobionate

OBJECTIVE: To determine whether inflammation of the jejunum of horses decreases the number of motilin receptors and amounts of motilin receptor mRNA and alters erythromycin lactobionate binding affinity to the motilin receptor in jejunal tissues. SAMPLE POPULATION: Jejunal segments in 6 adult horses. PROCEDURE: Each horse was anesthetized, and a ventral median celiotomy was performed; 2 segments of jejunum underwent a sham operation, 2 segments underwent ischemic strangulation obstruction (ISO), and 2 segments underwent intraluminal distension (ILD). Treatments were maintained for 120 minutes. From each segment, full-thickness biopsy samples were collected and smooth-muscle homogenates were prepared. Affinity and distribution of motilin binding to these preparations were determined by use of iodine 125 (125I)-labeled synthetic porcine motilin. Via displacement experiments, competition between 125I-labeled motilin and erythromycin lactobionate for binding to motilin receptors in the different segments was investigated. A quantitative real-time PCR technique was used to assess motilin receptor mRNA content in the muscle preparations. RESULTS: Compared with the ISO or ILD segments, the number of motilin receptors was significantly higher in the sham-operated segments; ILD segments contained the lowest number of motilin receptors. The expression of motilin receptor mRNA was significantly decreased in ILD segments but not in ISO segments. Erythromycin lactobionate displacement of 125I-labeled motilin from motilin receptors did not differ significantly among the jejunal segments. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that downregulation and decreased production of motilin receptors in inflamed jejunal tissue contribute to the altered prokinetic response to erythromycin in horses with gastrointestinal disease.     

Expression of the ether-a-go-go (ERG) potassium channel in smooth muscle of the equine gastrointestinal tract and influence on activity of jejunal smooth muscle

OBJECTIVE: To determine whether ether-a-go-go (ERG) potassium channels are expressed in equine gastrointestinal smooth muscle, whether ERG channel antagonists affect jejunal muscle contraction in vitro, and whether plasma cisapride concentrations in horses administered treatment for postoperative ileus (POI) are consistent with ERG channels as drug targets. SAMPLE POPULATION: Samples of intestinal smooth muscle obtained from 8 horses free of gastrointestinal tract disease and plasma samples obtained from 3 horses administered cisapride for treatment of POI. PROCEDURE: Membranes were prepared from the seromuscular layer of the duodenum, jejunum, ileum, cecum, large colon, and small colon. Immunoblotting was used to identify the ERG channel protein. Isolated jejunal muscle strips were used for isometric stress response to ERG channel blockers that included E-4031, MK-499, clofilium, and cisapride. Plasma concentrations of cisapride were determined in 3 horses administered cisapride for treatment of POI after small intestinal surgery. RESULTS: Immunoblotting identified ERG protein in all analyzed segments of the intestinal tract in all horses. The selective ERG antagonist E-4031 caused a concentration-dependent increase in jejunal contraction. Clofilium, MK-499, and cisapride also increased jejunal contraction at concentrations consistent with ERG channel block; effects of E-4031 and cisapride were not additive. Peak plasma cisapride concentrations in treated horses were consistent with ERG block as a mechanism of drug action. CONCLUSIONS AND CLINICAL RELEVANCE: The ERG potassium channels modulate motility of intestinal muscles in horses and may be a target for drugs. This finding may influence development of new prokinetic agents and impact treatment of horses with POI.     

Motility of the equine gastrointestinal tract: Physiology and pharmacotherapy

Normal gastrointestinal (GI) motility patterns are necessary to maintain transit of ingesta and to facilitate digestion and absorption of nutrients. Disorders of the equine GI tract are frequently encountered by the equine practitioner and these disorders are often associated with an interruption in normal intestinal motility patterns, thus complicating treatment of the primary disease. Consequently, numerous treatments have been investigated in horses to facilitate the return of normal intestinal motility. The purpose of this article is to provide a brief review of the anatomy and physiology of the GI tract in the horse and review medications available to the equine veterinarian that may potentially promote intestinal motility.     

Digestion of hay protein in different segments of the equine digestive tract

Mature ponies fitted with permanent ileal cannulas were used in a 3 X 3 Latin square experiment to quantify prececal, postileal and total tract digestion of hay protein. Coastal Bermuda grass (CB), containing 11.7% crude protein, and two qualities of alfalfa, containing 15.0% (low-protein alfalfa; LA) and 18.1% (high-protein alfalfa; HA) crude protein, were fed in coarsely chopped form at 2% of body weight daily. Total tract apparent digestibility of the N in HA (73.8%) was higher than that in CB (57.0%; P less than .05) and was slightly higher than that in LA (66.1%; P less than .10). Nitrogen in LA was apparently more digestible than that in CB (P less than .05). Apparent prececal digestibilities of N in LA and CB were 1.3% and 9.6%, respectively, and were lower (P less than .05), or tended to be lower (P less than .10), than the 21.0% observed for HA. In relative terms, an average of 9.4% of the total N digestion occurred in the upper tract when CB and LA were fed, whereas 28.5% of total N digestion occurred in the foregut when HA was fed. There was a slightly higher concentration of total plasma free amino acids (P less than .10) at 1 h postfeeding when horses were fed alfalfa. Also, N retention was higher when ponies were fed HA (P less than .05) than when LA or CB were fed. Apparent postileal N digestibility was 52.5% for CB, 65.7% for LA and 66.9% for HA. Differences were not significant, and the large intestine appeared to compensate for the inefficiency of N digestion in the upper tract.(ABSTRACT TRUNCATED AT 250 WORDS)     

Arteriovenous differences for glutamine in the equine gastrointestinal tract.

Glutamine has been shown to be an important metabolic substrate of enterocytes in many animals, including cats, dogs, hamsters, human beings, monkeys, rabbits, rats, and sheep. To determine whether glutamine is important in the metabolism of cells of the equine gastrointestinal tract, we examined transintestinal differences in glutamine concentrations in the arterial and venous circulation, and measured activity of the major glutamine catabolizing enzyme, glutaminase. Arteriovenous differences provide an index of the amount of a given substrate removed by the tissue across which the measurements are made, and commonly are expressed as a percentage of substrate removed, or percent extraction. Arteriovenous differences for glutamine were determined in 7 anesthetized adult horses (weight, 450 to 500 kg) before and after an i.v. glutamine infusion. The mean baseline arterial glutamine concentration (+/- SEM) was 572 +/- 24 microM; this concentration quadrupled (to 2,167 +/- 135 microM, P less than 0.01) 1 minute after i.v. bolus infusion of a 17.5-g glutamine load. Baseline extraction by the portal-drained viscera was 7.5 +/- 1.5%; this value increased to 18 +/- 2% at 1 minute (P less than 0.01) and had returned to baseline values 60 minutes later. Arteriovenous differences were greatest across the jejunum (11.8 +/- 1.8% in the baseline period vs 33.1 +/- 3.1% at 1 minute, P less than 0.001), with smaller differences across the colon, suggesting that the jejunum was the more avid utilizer of glutamine. Glutaminase activity was 4.38 +/- 0.16 and 4.00 +/- 0.60 mumol/mg of protein/h under standard conditions in jejunal and ileal mucosa, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Distribution of the neurokinin-1 receptor in equine intestinal smooth muscle

REASON FOR PERFORMING STUDY: Tachykinins have profound effects on equine intestinal motility, but the distribution of the neurokinin receptors (NKRs) through which they act is unknown. This study reports the distribution of one of these receptors, the neurokinin-1 receptor (NK1R), in smooth muscle throughout the equine intestinal tract. OBJECTIVES: To quantify the distribution of the NK1R, based upon mRNA expression, in smooth muscle of different regions of the equine intestinal tract. METHODS: Nine regions of the intestinal tract were sampled in 5 mature horses. Total RNA was isolated from smooth muscle and reverse transcribed; NK1R mRNA was then quantified using real-time PCR. RESULTS: NK1R mRNA was found at all levels of the sampled intestinal tract. The smooth muscle of the proximal small intestine and the ventral colon exhibited the highest level of NK1R mRNA expression in the equine intestinal tract. CONCLUSIONS: Tachykinins probably affect intestinal contractility and propulsion in the proximal small intestine and in the ventral colon.     

Characterization of alpha-adrenoceptor subtypes in smooth muscle of equine ileum

OBJECTIVE: To determine the concentration and binding characteristics of alpha-adrenoceptor subtypes in smooth muscle cell membranes of equine ileum. SAMPLE POPULATION: Segments of longitudinal and circular smooth muscle from the ileum of 8 male and 8 female adult horses. PROCEDURE: Distribution of alpha-adrenoceptor subtypes was assessed by use of radioligand binding assays incorporating [3H]-prazosin and [3H]-rauwolscine, highly selective alpha1- and alpha2-adrenoceptor antagonists, respectively. Characterization of adrenoceptor subtypes was performed by use of binding inhibition assays. RESULTS: On the basis of binding affinity for specific radioligands, low- and high-affinity alpha1- and alpha2-adrenoceptors were detected. Concentration of low-affinity alpha2-adrenoceptors was significantly greater in male horses, compared with females. Competition studies confirmed the specificity of the radioligands used in the binding assays. Alpha1-adrenoceptors of both subtypes in male and female horses had a higher affinity for prazosin than phentolamine, whereas yohimbine did not compete with the radioligand for binding. For alpha2-adrenoceptors regardless of subtype, potency of inhibition elicited by each drug varied between sexes. In males, yohimbine was a more potent inhibitor than phentolamine, which was more potent than prazosin. In females, yohimbine was more potent than prazosin, which was more potent than phentolamine. CONCLUSIONS AND CLINICAL RELEVANCE: High- and low-affinity alpha1- and alpha2-adrenoceptors were detected in smooth muscle of equine ileum. Because alpha-adrenoceptor subtypes, particularly alpha2-adrenoceptors, are involved in the regulation of gastrointestinal tract function, characterization of these receptors may represent the basis for development of new therapeutic strategies for the control of gastrointestinal disturbances in horses.     

Characterization of the in vitro responses of equine cecal longitudinal smooth muscle to endothelin-1

OBJECTIVE: To characterize the in vitro response of equine cecal longitudinal smooth muscle (CLSM) to endothelin (ET)-1 and assess the role of ETA and ETB receptors in those ET-1-induced responses. ANIMALS: 36 horses without gastrointestinal tract disease. PROCEDURE: To determine cumulative concentration-response relationships, CLSM strips were suspended in tissue baths containing graded concentrations of ET-1 (10(-9) to 10(-6)M) with or without BQ-123 (ETA receptor antagonist); with or without IRL-1038 (ETB receptor antagonist); or with both antagonists at concentrations of 10(-9), 10(-7), and 10(-5)M. To determine the percentage change in baseline tension of CLSM, the areas under the curve during the 3-minute periods before and after addition of each dose were compared. Also, the effects of ET-1 and a combination of selective ETA and ETB receptor antagonists on electrically evoked contractions were studied. RESULTS: ET-1 caused sustained increases in CLSM tension in a concentration-dependent manner. Contractile responses to ET-1 were not significantly inhibited by either BQ-123 or IRL-1038 alone at any concentration; however, responses were significantly inhibited by exposure to the antagonists together at a concentration of 10(-5)M. Electrical field stimulation did not change the spontaneous contractile activity of CLSM and did not significantly alter the tissue response to ET-1, BQ-123, or IRL-1038. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that ET-1 has a contractile effect on equine CLSM that is mediated via ETA and ETB receptors. In vitro spontaneous contractions of equine CLSM apparently originate in the smooth muscle and not the enteric nervous system.     

Use of sugar beet in porcine nutrition. 1. Effect of various methods of treatment on the gastrointestinal tract

The influence of the size of particles, of steaming and preserving (formic acid) of the sugar beet 'Rosamona' was studied with respect to apparent digestibility and tract parameters. Digestibility measuring only resulted in an improvement for crude protein due to the cutting up into small pieces. Tract measures and fillings, microbial activity and an estimate of the absorption rates of nutrients indicate that the bigger part of fresh and silaged coarse particles is digested in the colon but that the nutrients of steamed (and partly mashed) beets are predominantly pre-caecally digested.     

Passage rate of digesta through the equine gastrointestinal tract: A review

Passage rate through the equine gastrointestinal tract is best described by mean retention time (MRT). Transit times of digesta differ greatly between the different parts of the gastrointestinal tract. Passage through the stomach and small intestine is rapid (5 h on average), whereas a longer retention time is recorded in the cecum and colon (35 h on average).Important considerations in estimating MRT have been the use of a solute phase and a fluid phase marker and calculations applied, especially when sampling frequency is reduced.A number of animal and feed related factors influence MRT: body weight, pregnancy and lactation may increase passage rate, whereas the impact of exercise, the most important animal related factor, will depend on the exercise type and the fluid/particle ratio.Information on the effect of the main feed related factors is lacking. Smaller particles and feed with a higher water-holding capacity move slower through the gut, but reduced fibre length, increased feeding level and increased forage/concentrate ratio will accelerate passage rate. Feeding frequency seems not to affect passage rate.Sometimes, conclusions are contradictory as effect of animals, feeds and methods are confusing.     

Survival and colonization of orally administered Lactobacillus plantarum 301102 in porcine gastrointestinal tract

The purpose of this study was to evaluate the survival and colonization of Lactobacillus plantarum 301102 in porcine gastrointestinal tract and its influence on the intestinal microflora. This strain was isolated from traditional cheese from Inner Mongolia, China. Fermented milk prepared with strain 301102 was administered to pigs and fecal microflora was examined. Intestinal passage of strain 301102 was monitored by a combination of selection with selective medium and the carbohydrate fermentation test, and confirmed by analysis of plasmid DNA patterns. Colonization of this strain was assessed by recovery 7 days after administration. The numbers of Lactobacillus and Bifidobacterium cells in the feces of pigs administered fermented milk prepared with strain 301102 were increased. This strain was recovered from all the pigs during the administration period, and from four of six pigs 7 days after administration. L. plantarum 301102 can pass through the gastrointestinal tract, balance the intestinal microflora, and colonize the intestinal tract, and is therefore an appropriate candidate for a probiotic.     

Effects of verapamil on the smooth muscle of the horse urinary tract

The effects of verapamil, a calcium antagonist agent, were studied on smooth muscle preparations of the lower urinary tract of horses. Verapamil (2 X 10(-4) to 2 X 10(-8) M) relaxed the ureter, urethra and urinary bladder preparations contracted by potassium (127 mM), L-noradrenaline (2 X 10(-5) M), histamine (2 X 10(-5) M) and acetylcholine (2 X 10(-5) M). These results allow the conclusion that verapamil has a dose-dependent relaxing effect on smooth muscle of the lower urinary tract.     

Epitheliotropic T-cell gastrointestinal tract lymphosarcoma with metastases to lung and skeletal muscle in a cat

An Abyssinian cat examined because of hypoproteinemia and weight loss was found to have epitheliotropic T-cell lymphosarcoma of the gastrointestinal tract. Neoplastic lymphocytes infiltrated the epithelium and lamina propria of the stomach, small intestine, and large intestine. Metastatic foci were found in the lungs and the musculature of the right hind limb. Chemotherapy resulted in transient shrinkage of the lung and limb masses; however, these masses grew to approximately their original size within 2 weeks after initiation of treatment, and the cat was euthanatized.     

An immunohistochemical study of interstitial cells of Cajal (ICC) in the equine gastrointestinal tract.

The interstitial cells of Cajal (ICC) are c-kit immunoreactive cells of the gastrointestinal tract which are suggested to have a role in the control of intestinal motility. Cells with c-kit immunoreactivity have not been previously described in the gastrointestinal tract of the horse. Immunoreactivity for c-kit was revealed using immunohistochemical labelling with an anti-c-kit polyclonal antibody. Sections of normal gastrointestinal tissue were examined from 13 anatomically defined sites from stomach to small colon taken from horses free from gastrointestinal disease. Three types of c-kit immunoreactive cells were identified: spindle-shaped cells in the region of the myenteric plexus, stellate or bipolar cells in the circular muscle layer, and round cells in the submucosa. The round cells were shown to be mast cells with the use of toluidine blue staining, whereas the other c-kit immunoreactive cells did not exhibit metachromasia and were classified as ICC. This study will serve as a basis for future pathological studies in the horse.