Rabbit Intestine Research Articles

Resolution enhancement of pump–probe microscope with an inverse-annular filter

Optical pump–probe microscopy can provide images by detecting changes in probe light intensity induced by stimulated emission, photoinduced absorbance change, or photothermal-induced refractive index change in either transmission or reflection mode. Photothermal microscopy, which is one type of optical pump–probe microscopy, has intrinsically super resolution capability due to the bilinear dependence of signal intensity of pump and probe. We introduce new techniques for further resolution enhancement and fast imaging in photothermal microscope. First, we introduce a new pupil filter, an inverse-annular pupil filter in a pump–probe photothermal microscope, which provides resolution enhancement in three dimensions. The resolutions are proved to be improved in lateral and axial directions by imaging experiment using 20-nm gold nanoparticles. The improvement in X (perpendicular to the common pump and probe polarization direction), Y (parallel to the polarization direction), and Z (axial direction) are by 15 ± 6, 8 ± 8, and 21 ± 2% from the resolution without a pupil filter. The resolution enhancement is even better than the calculation using vector field, which predicts the corresponding enhancement of 11, 8, and 6%. The discussion is made to explain the unexpected results. We also demonstrate the photothermal imaging of thick biological samples (cells from rabbit intestine and kidney) stained with hematoxylin and eosin dye with the inverse-annular filter. Second, a fast, high-sensitivity photothermal microscope is developed by implementing a spatially segmented balanced detection scheme into a laser scanning microscope using a Galvano mirror. We confirm a 4.9 times improvement in signal-to-noise ratio in the spatially segmented balanced detection compared with that of conventional detection. The system demonstrates simultaneous bi-modal photothermal and confocal fluorescence imaging of transgenic mouse brain tissue with a pixel dwell time of 20 µs. The fluorescence image visualizes neurons expressing yellow fluorescence proteins, while the photothermal signal detected endogenous chromophores in the mouse brain, allowing 3D visualization of the distribution of various features such as blood cells and fine structures most probably due to lipids. This imaging modality was constructed using compact and cost-effective laser diodes, and will thus be widely useful in the life and medical sciences. Third, we have made further resolution improvement of high-sensitivity laser scanning photothermal microscopy by applying non-linear detection. By this, the new method has super resolution with 61 and 42% enhancement from the diffraction limit values of the probe and pump wavelengths, respectively, by a second-order non-linear scheme and a high-frame rate in a laser scanning microscope. The maximum resolution is determined to be 160 nm in the second-order non-linear detection mode and 270 nm in the linear detection mode by the PT signal of GNPs. The pixel rate and frame rate for 300 × 300 pixel image are 50 µs and 4.5 s, respectively. The pixel and frame rate are shorter than the rates, those are 1 ms and 100 s, using the piezo-driven stage system.

Immunobiological and morphofunctional indicators with dysbacteriosis of the intestine of rabbits

The article presents results of research of dynamics of morphological, biochemical, haematological and immunological parameters when changing the quantity and species of microorganisms of intestinal microbiocenosis of the rabbits. With the dominance of pathogenic bacteria in rabbit intestinal microbiocenoses, the titer of bifidumbacteria was 10-5-10-4, a decrease in the number of lactobacilli, an increase in the number of enterobacteria, a correlation between the colonization index and the degree of adhesion of bacteria was revealed; an increase in the total number of leukocytes, lymphocytes, erythrocytes, hematocrit, alanine aminotransferase level, aspartate aminotransferase, lactate dehydrogenase, total bilirubin, -amylase, albumin, alkaline phosphatase, T-killer level, NK cells, concentration of C-reactive protein. We observed a decrease in the concentration of hemoglobin, monocytes, total protein level, T-lymphocytes, T-helpers, B-lymphocytes, phagocytic index, phagocytic activity of leukocytes, NST-spontaneous, NST-stimulated. Dynamics of morphological indicators was characterized by a disorder of the circulation of organs, a predominance of dystrophic and necrotic processes in the tissues and organs of the cardiovascular, respiratory, digestive, excretory system. Infiltration with mononuclear and segmented leukocytes was noted, diffuse lymphoid proliferation of the mucous membranes of the organs, hyperemia, edema, multiple hemorrhages, accumulation of hemorrhagic exudate in the lumen of the respiratory, digestive and urogenital tracts. Observed catarrhal-hemorrhagic inflammation of the gastrointestinal tract; mucous membrane of the stomach, small and large intestines edematic, hyperemic, with hemorrhages, covered with mucus, fibrinous films, with necrotic areas. The signs of acute diffuse interstitial nephritis, characterized by proliferation of connective tissue cells with a predominance of lipoblast and epithelioid cells, are revealed. In the presence of bacterial cells in the lumen of the tubules, the parenchyma of the cortical and medullar layers was marked by microabscesses. Accidental transformation of the thymus was observed, characterized by signs of a circulatory disorder, a decrease in the area of the cortex, the formation of cysts; signs of diffuse hyperplasia of the spleen with necrobiotic changes in the foci of proliferation, reduction of the area of lymphoid follicles, expansion of the area occupied by the red pulp of follicles, hyperplasia of follicles of white pulp with foci of necrosis. Identified multiple bacterial embolism of blood vessels, developed signs of hemorrhagic Diathesis, watched the development of sign infiltrative itch processes.

Regional and mucosal distributions of some intestinal immunoreactive endocrine cells in New Zealand white rabbit (Oryctolagus cuniculus L.).

The aim of this study was to detect the regional and mucosal distribution of endocrine cells that secrete gulcagon, somatostatin, Chyholecystokinin-8 (CCK-8), serotonin, secretin, substance P (SP) and histamine in the small and large intestine of New Zealand white rabbit (Oryctolagus cuniculus L.) using immunohistochemical peroxidase-antiperoxidase (PAP) method. It was found that most of the immunoreactive (IR) endocrine cells, which are oval- or spindle-shaped, are spotted in the basal parts of the relevant glands. It was noticed that cells in the lamina epithelialis of small and large intestine is linked to the lumen and that the cells in their glands cannot reach the lumen. Immunoreactive cells for glucagon, somatostatin, serotonin, secretin and SP were identified in lamina epithelialis of the small and large intestine. It was seen that secretin, SP and histamine-IR cells are rarely deployed throughout the intestinal tract. It was determined that somatostatin-IR cells were identified throughout the intestinal tract. In conclusion, the immunohistochemical study shows that gastrointestinal tract of this species contained different types of endocrine cells similar to those found in other vertebrate species. However, some species-dependent unique distributions and frequencies of endocrine cells were also observed in the present study.

Unique regulation of Na-glutamine cotransporter SN2/SNAT5 in rabbit intestinal crypt cells during chronic enteritis.

The only Na‐nutrient cotransporter described in mammalian small intestinal crypt cells is SN2/SNAT5, which facilitates glutamine uptake. In a rabbit model of chronic intestinal inflammation, SN2 stimulation is secondary to an increase in affinity of the cotransporter for glutamine. However, the immune regulation of SN2 in the crypt cells during chronic intestinal inflammation is unknown. We sought to determine the mechanism of regulation of Na‐nutrient cotransporter SN2 by arachidonic acid metabolites in crypt cells. The small intestines of New Zealand white male rabbits were inflamed via inoculation with Eimeria magna oocytes. After 2‐week incubation, control and inflamed rabbits were subjected to intramuscular injections of arachidonyl trifluoromethyl ketone (ATK), piroxicam and MK886 for 48 hrs. After injections, the rabbits were euthanized and crypt cells from small intestines were harvested and used. Results: Treatment of rabbits with ATK prevented the release of AA and reversed stimulation of SN2. Inhibition of cyclooxygenase (COX) with piroxicam did not affect stimulation of SN2. However, inhibition of lipoxygenase (LOX) with MK886, thus reducing leukotriene formation during chronic enteritis, reversed the stimulation of SN2. Kinetic studies showed that the mechanism of restoration of SN2 by ATK or MK886 was secondary to the restoration of the affinity of the cotransporter for glutamine. For all treatment conditions, Western blot analysis revealed no change in SN2 protein levels. COX inhibition proved ineffective at reversing the stimulation of SN2. Thus, this study provides evidence that SN2 stimulation in crypt cells is mediated by the leukotriene pathway during chronic intestinal inflammation.

Histochemical and biochemical study of rabbit intestine in healthy and affected by epizootic enteropathy animals

Rabbits are suffering from Epizootic Rabbit Enteropathy (ERE), a new gastro-intestinal syndrome of unknown aetiology. ERE has not yet been investigated thoroughly from the patho-physiological point of view of the intestine malfunction during the intestine dysbiosis. For this reason, the aim of the present research was to study the haematological profile, the intestine histopathology and certain biochemical parameters (α-amylase activity, blood glucose concentration) of animals affected by ERE in comparison with those of healthy rabbits of the same age. In six healthy and six, affected by ERE, crossbreed New Zealand X California 35-day old rabbits, the following analyses were performed: Total blood count, histopathological morphology of the intestine, alkaline phosphatase (ALP) distribution in intestine brush border and α-amylase activity in blood plasma and small intestine epithelium, respectively. Total blood count showed significantly (at 95% confidence level) decreased values for all blood parameters except for the white blood cell count, which proved to be significantly higher compared in ERE rabbits to that of normal rabbits. The a-amylase activity and concentration in blood plasma and intestinal epithelium were significantly (a=0.05) decreased, in contrast to blood glucose concentration, that was found to be significantly increased in ERE rabbits. Stomach was full of watery content, intestine presented non-specific enteropathy and mainly the small intestine was full of gas and watery content. Caecal and colon presented impaction and mucus was present in the colon. The histopathological evaluation of the ileum, sacculus rotundus, caecum and colon presented, mainly, lamina propria mononuclear cell infiltration and swelling, vacuolation, flattening and denuding of the enterocytes as well as oedematous lymphoid tissues. Duodenum had necrotic villi and deep infiltration with mononuclear and polymorphonuclear cells, within the lamina propria. Also swelling, vacuolation, flattening, enterocytes denuding and oedematous lymphoid tissue were observed. Jejunum had no lesions. The caecum and the colon presented an ALP positive reaction along the brush border of the epithelial cells. The small intestine presented a positive reaction along the brush border of intestinal glands-upper part- cells only and the bases of a few of the villi.

Morphometric parameters of the intestine and aggregated lymphatic nodules of meat rabbits

Nowadays, new breeds of rabbits are appearing with high productivity and rapid growth rate. These breeds require fresh research to be conducted in different fields, which include morphology of the intestine and gut-associated lymphoid tissue (GALT). The morphology of the intestine and GALT of meat rabbits are the investigated objects of the current research. We conducted the investigation on rabbits of different ages (1, 10, 20, 30, 60 and 90 days old). Each age group consisted of five animals. All veterinary ethical requirements were complied with. The body lengths and body masses were measured. Using t-coefficients, we estimated the range of variation of these parameters in each age group. The conclusion about this variation was taken. The masses and lengths of the different intestine parts were investigated as well. Using the obtained values, we provided a continuous numerical model to compute the mass and length of any intestine part of a rabbit of any age in the range from one to 90 days. This allowed us to represent the obtained empirical model by a plot for subsequent detailed study. We found that the changes in masses and lengths do not follow the same pattern for all intestine parts. For some intestine parts, this pattern is approximately linear in the range from one to 90 days; meanwhile for others, the changes in masses and lengths are quite complicated. We observed two local maximas of the growth rate for some intestine parts. Apart from the masses and lengths of the intestine parts, we accomplished macro analysis of the aggregated lymphoid nodules of each age group of animals. We estimated the index of gross area and the law by which it changes in each group. The research results presented here may be used as a basis for composing food ration, assessment of immune status and development of effective prevention and treatment against some diseases.

Intestinal Mechanomorphological Remodeling Induced by Long-Term Low-Fiber Diet in Rabbits

Short-term feeding with low-fiber diet remodels the mechanomorphological properties in the rabbit small intestine. The aims were to study the effect of feeding low-fiber diet for 5months on mechanomorphological properties including the collagen fraction in the rabbit intestines. Fifteen rabbits were divided into an Intervention group (IG, n=10) fed a low-fiber diet and a Control group (CG, n=5) fed a normal diet for 5 months. Five months later, four 10-cm-long segments obtained from the duodenum, jejunum, ileum and large intestine were used for histological and mechanical analysis, respectively. The wall thickness, wall area, mucosa and muscle layer thickness decreased whereas the submucosa layer thickness increased in the IG (p<0.05). The collagen fraction decreased in all layers and segments in the IG (p<0.05). The opening angle increased in the large intestine and decreased in the ileum in the IG (p<0.05). The intestinal stress-strain curves for IG shifted to the right, indicating softening. The creep did not change in the four segments. The wall stiffness was associated with wall thickness and collagen fraction in the submucosa layer. Long-term low-fiber diet in rabbits induced histomorphometric and biomechanical remodelling of the intestines.

Recombinant Lactobacillus casei expressing Clostridium perfringens toxoids α, β2, ε and β1 gives protection against Clostridium perfringens in rabbits

The present study used Lactobacillus casei ATCC 393 as antigen delivery system to express C. perfringens toxoids α-β2-ε-β1 to construct the recombination Lactobacillus casei pPG-2-α-β2-ε-β1/L. casei 393. After being induced by 1% xylose, the specificity and integrity of recombinant strain were determined by Western-blotting. Rabbits as native animal model were immunized orally with pPG-2-α-β2-ε-β1/L. casei 393 and the titers of specific IgG and sIgA were determined by ELISA. The result showed that oral administration with the recombinants could elicit both local mucosal and systemic immune responses. The proliferation of spleen lymphocytes in rabbits immunized with pPG-2-α-β2-ε-β1/L. casei 393 was observed. Levels of IL-4 and IFN-γ produced were significantly higher in lymphocytes isolated from the vaccine group than those from the control groups. Flow cytometry assay showed that both the percentages of CD4+T cells and CD8+T cells from the vaccine group were significantly increased than the control groups. All these results showed that immunizing with recombinants can elicit both humoral immunity and cellular immunity. Besides, in order to determine the effectiveness of oral immunization with pPG-2-α-β2-ε-β1/L. casei 393, rabbits of vaccine group and control groups were challenged with 1×LD100 unit of culture filtrate of C. perfringens type C and type D toxins respectively. After challenge, 100% of the immunized rabbits survived, while the rabbits of the control group were killed within 48h. Observation on histopathology showed that histopathological changes were obviously found in heart, liver, spleen, lung, kidney, intestine and brain of rabbits from the control groups, while no apparent histopathological change was observed in the vaccine group. All the results show that pPG-2-α-β2-ε-β1/L. casei 393 can eliciteffective immunoprotection against C. perfringens. All of these suggest that the use of pPG-2-α-β2-ε-β1/L. casei 393 can be regarded as candidate for the development of a vaccine against C. perfringens.

Tri-block co-polymer nanocarriers for enhancement of oral delivery of felodipine: preparation, in vitro characterization and ex vivo permeation

This study aimed to prepare, optimize and characterize novel felodipine-loaded polymeric nanomicelles, using a pluronic mixture of F127 and P123. Thin-film hydration method was adopted for the preparation of different polymeric nanomicelles (T1–T12) according to a 41.31 full factorial design. Factors studied were: Pluronic®:drug ratio (P:D ratio) (10, 20, 30 and 40 w/w) and percent of hydrophilic polymer (F127%) (33.33%, 50% and 66.67% w/w). Optimization criteria were to maximize transmittance percent (T%) and entrapment efficiency percent (EE%) and to minimize particle size (PS) and polydispersity index (PDI). The optimized formulation was further characterized by DSC, FTIR and 1H NMR studies. It was also subjected to stability testing and ex vivo permeation using rabbit intestines. Spherical nanomicelles of particle size ranging from 26.18 to 87.54 nm were successfully obtained. The optimized formulation was found to be the already prepared formulation T12 (P:D ratio of 40 and 66.67% F127) with suitable T% and EE% of 95.12% and 91.75%, respectively. DSC, FTIR and 1H NMR studies revealed felodipine (FLD) incorporation within T12 nanomicelles. T12 enhanced the ex vivo intestinal permeation of FLD when compared to a drug suspension and showed good stability. Therefore, pluronic nanomicelles could be promising for improved oral delivery of FLD.

The influence of food restriction on the small bowel: Does intensive short‑term food restriction lead to weight loss?

The objective of this study was to show morphological changes in the small intestine of Hyplus broiler rabbits following an eight‑day food restriction programme. The control group (C) received food ad libitum (ADL) for the duration of the experiment. Group R1 received 50g of food per day, and group R2 received 65g of food per day. After the food restriction diet had been completed, groups R1 and R2 were returned to ad libitum feeding. After food restriction and at the end of the experiment, the longest small bowel measurement was recorded in the C group. In the C group, after food restriction, the villi height was significantly higher, compared to that in R1 and R2 groups and at the end of the experiment, the villi were significantly higher in R1 and R2 groups. After food restriction, the values of crypts depth were approximately similar in all groups, and the end of experiment, the depth of crypts were deepest in R1and R2 groups, as compared to that in C group. The full process is followed by weight loss to the end of the experiment. These data suggest that intensive short‑term food restriction followed by ADL feeding has effect on weight loss (Fig. 3, Ref. 30).

Resolution enhancement of pump-probe microscopy with an inverse-annular spatial filter

We have introduced a pupil filter, an inverse-annular pupil filter in a pump-probe photothermal microscope, which provides resolution enhancement in three dimensions. The resolution is probed to be improved in lateral and axial resolution by imaging experiment using 20 nm gold nanoparticles. The improvement in X (perpendicular to the common pump and probe polarization direction), Y (parallel to the polarization direction), and Z (axial direction) are by 15±6, 8±8, and 21±2 % from the resolution without a pupil filter. The resolution enhancement is even better than the calculation using vector field, which predicts the corresponding enhancement of 11, 8, and 6 %. The discussion is made to explain the unexpected results. We also demonstrate the photothermal imaging of thick biological samples (cells from rabbit intestine and kidney) stained with hematoxylin and eosin dye with the inverse-annular filter.

WITHDRAWN: Identifying differentially expressed genes in the intestine of healthy and diarrheal Rex Rabbits by RNA-Seq

WITHDRAWN: Identifying differentially expressed genes in the intestine of healthy and diarrheal Rex Rabbits by RNA-Seq

Compost-sourced substances (SBO) as feedstuff additives in rabbit production

Acid soluble bio-organic (SBO) substances have been extracted from plant bio-waste. The SBOs have been tested as feed additives for reared animals. Two trials (T1 and T2, lasting 63 and 56 days, respectively) were carried out to evaluate the effect of different doses of SBOs as feed additives in diets for fattening rabbits (131 and 120 animals from 35 days of age, respectively) and as binders for the manufacturing of diet pellets to improve their physical properties. The effect of different SBO doses (0, 0.5, 2.5g/kg for T1 and 0, 5, 10g/kg for T2) on the growth performance, health status, diet digestibility, carcass and meat traits of rabbits was studied. The aggregating effect of SBOs on feed particles during pelleting was studied by adding 50 and 100g/kg of SBOs to a feed, and then measuring the shear and compression force on the pellets. The results have shown that SBO supplementation of rabbit diets at a concentration of 0.5–10g/kg does not affect the live and slaughtering performances, except for the intestinal tract absorption area (higher in the intestines of rabbits fed with a high supplementation of SBO; P<0.05) or the dry matter, organic matter, ether extract, fibre and gross energy apparent digestibility of the feed (lower in diets with low or no supplementation of SBO; P<0.05). The rabbits fed with the lower SBO dose showed higher values of total anaerobic bacteria and Bacteroides (P<0.05) and a higher cadmium meat content (P<0.05) in T1 and T2 than all others groups. SBOs included at concentrations of 10 and 20% were also found to affect the physical properties of the pelleted feed. These pellets are more resistant to shear (P<0.05) and compression force (P<0.01) in the presence of added SBO. It can be concluded that SBO may be used as a feed additive for rabbits, without any adverse effects on animal productivity and health, as well as to improve the physical properties of pelleted feeds.

Laxative Effect of Some Plants Extracts Used in Traditional Pharmacopoeia on the Intestine of Rabbit

Constipation is a condition in which a person or animal has difficulty in eliminating solid waste from the body and the faeces are hard and dry. It is mainly characterized by a difficulty in defecation. The aim of the study is to assess the laxative effect of some plants such as Adansonia digitata , Ziziphus mauritiana and Psidium guayava traditionally used in Chad to treat the problems of constipation. The Microdynamometer method (Biomecatronics) is used to carry out this study. The outcomes show that the extracts’ leaves of Psidium guayava and barks of Ziziphus mauritiana did not lead a significant contraction of duodenum of rabbit until a maximum concentration of 1000 μg, but the extracts’ leaves of Adansonia digitata had a significant effect (P Adansonia digitata varies with the increase of its concentration (dose dependent) with a maximum of 500 μg and the minimal effect is inferior to 10 μg. On the other hand, the extract of Adansonia digitata has a musculotrope effect because the results have shown that inhibitive molecules such as the atropine and the naloxone do not have to inhibit the action of Adansonia digitata on the intestinal motility of rabbit. Thus, the extract of this plant would reduce significantly the time of intestinal transit and it will be an important laxative drug candidate as described traditionally in Chad.

Histomorphological Relationship of Paneth Cells with Stem Cells in the Small Intestine of Indigenous Rabbit at Different Postnatal Ages

Histomorphological Relationship of Paneth Cells with Stem Cells in the Small Intestine of Indigenous Rabbit at Different Postnatal Ages

Mechanisms Of Fructose Absorption

Mechanisms Of Fructose Absorption

Ultrastructural Changes in the Intestine of Suckling Rabbits Infected with Cholerogenic and Non-Cholerogenic nonO1/nonO139 Vibrio cholerae Strains.

We performed an electron microscopic study of the small intestine of suckling rabbits infected with cholerogenic and non-cholerogenic strains nonO1/nonO139 Vibrio cholerae. Cholerogenic strain induced mostly hydropic degeneration of the epithelium typical of cholera toxin effect, while non-cholerogenic strain induced the formation of lacunae along the borders of adjacent epithelial cells typical of hemagglutinin/protease effect. In both cases, reduction of microvilli, destruction of intracellular organelles, two types of mitochondrial reaction (condensation and swelling with destruction of cristae), appearance of myelin figures, defects in the capillary walls, and activation of pinocytosis were observed. These data confirm our previous assumption on interchangeability of different pathogenic factors of Vibrio cholerae, including nonO1/nonO139 strains.

Infant Rabbit Model for Diarrheal Diseases.

Vibrio cholerae is the agent of cholera, a potentially lethal diarrheal disease that remains a significant threat to populations in developing nations. The infant rabbit model of cholera is the only non-surgical small animal model system that closely mimics human cholera. Following orogastric inoculation, V. cholerae colonizes the intestines of infant rabbits, and the animals develop severe cholera-like diarrhea. In this unit, we provide a detailed description of the preparation of the V. cholerae inoculum, the inoculation process and the collection and processing of tissue samples. This infection model is useful for studies of V. cholerae factors and mechanisms that promote its intestinal colonization and enterotoxicity, as well as the host response to infection. The infant rabbit model of cholera enables investigations that will further our understanding of the pathophysiology of cholera and provides a platform for testing new therapeutics.

Comparison of Flurbiprofen Tablets Available In Pakistani Market and Their Absorption Studies

AbstractThe aim of this present work was to compare different parameters of various brands of flurbiprofen tablets collected from different retail pharmacies in the local market of Pakistan. Four brands A, B, C and D were tested for weight variation, hardness, friability, disintegration dissolution, HPLC assay and in vitro absorption studies in rabbit skin, stomach and intestine by using the prepacked Column RT 250-4.6 Purospher® STAR RP-18 end capped (5 µm) and acetonitrile, phosphate buffer (pH 3.7) as mobile phase in the ratio of 1:1. Flurbiprofen was detected at 265 nm at the flow rate of 1 ml/min. Brand B was considered as reference. Similarity factor (f2) of brand B and C and brand B &amp; D was found to be 61 and 51 and dissimilarity factors (f1) values were 5 and 9 respectively in same dissolution medium. Model dependent methods First order, Hixon crowell and Weibull model were used. The method was found to be sensitive and linear in the range of 10 to 700 ppm with 0.999 coefficient of correlation. Everted sac absorption studies of selected formulation showed 50% of drug absorption from stomach in first 3 hours, 21% through intestine and very negligible through skin.

Mast cell regulation of Na-glutamine co-transporters B0AT1 in villus and SN2 in crypt cells during chronic intestinal inflammation.

BackgroundIn the chronically inflamed rabbit small intestine, brush border membrane (BBM) Na-glutamine co-transport is inhibited in villus cells (mediated by B0AT1), while it is stimulated in crypt cells (mediated by SN2/SNAT5). How mast cells, known to be enhanced in the chronically inflamed intestine, may regulate B0AT1 in villus and SN2/SNAT5 in crypt cell is unknown. Thus, the aim of the present study is to determine the regulation of B0AT1 and SN2/SNAT5 by mast cells during chronic enteritis.MethodsChronic intestinal inflammation was induced in male rabbits with intra-gastric inoculation of Eimeria magna oocytes. Rabbits with chronic inflammation were treated with ketotifen (10 mg/day) or saline (Placebo) for 2 days. Villus and crypts cells were isolated from the rabbit intestine using the Ca++ chelation technique. Na/K-ATPase activity was measured as Pi from cellular homogenate. BBM vesicles (BBMV) were prepared from villus and crypt cells and uptake studies were performed using rapid filtration technique with 3H-Glutamine. Western blot analyses were done using B0AT1 and SN2 specific antibodies.ResultsIn villus cells, Na-glutamine co-transport inhibition observed during inflammation was completely reversed by ketotifen, a mast cell stabilizer. In contrast, in crypt cells, Na-glutamine co-transport stimulation was reversed to normal levels by ketotifen. Kinetic studies demonstrated that ketotifen reversed the inhibition of B0AT1 in villus cells by restoring co-transporter numbers in the BBM, whereas the stimulation of SN2/SNAT5 in crypts cells was reversed secondary to restoration of affinity of the co-transporter. Western blot analysis showed that ketotifen restored immune-reactive levels of B0AT1 in villus cells, while SN2/SNAT5 levels from crypts cell remained unchanged.ConclusionIn the present study we demonstrate that mast cells likely function as a common upstream immune pathway regulator of the Na-dependent glutamine co-transporters, B0AT1 in villus cells and SN2 in crypts cells that are uniquely altered in the chronically inflamed small intestine.