Histoplanimetrical Study on the Relationship between Cellular Kinetics of Epithelial Cells and Proliferation of Indigenous Bacteria in the Rat Colon

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ABSTRACT. The relationship between the invasion of indigenous bacteria into intestinal crypts and the proliferation of epithelial cells was histoplanimetrically investigated in the rat ascending colon. Indigenous bacteria preferentially adhered to the intestinal superficial epithelial cells in the mesenterium-attached mucosa (MAM) compared to those in the mesenterium-non-attached mucosa (MNM). Intestinal crypts with indigenous bacteria were also significantly more frequently found in MAM than in MNM. Total epithelial cells, columnar epithelial cells and goblet cells were significantly more abundant in the intestinal crypts with no-indigenous bacteria in MAM (MAM-C) than those in MNM (MNM-C), whereas the columnar epithelial cells were less abundant in MAM-C than in the intestinal crypts with indigenous bacteria in MAM (MAM-C-B). Columnar epithelial cells and goblet cells immuno-positive for proliferating cell nuclear antigen (PCNA) in MAM-C were more abundant than those in MNM-C, but less abundant than those in MAM-C-B. Toll-like receptor (TLR)-2, -4 and -9 were immuno-positive in the striated borders of the intestinal superficial epithelial cells, but their positive intensities were weaker in MAM than in MNM. From these findings, indigenous bacteria were confirmed to preferentially settle on the intestinal superficial epithelium of MAM in the rat ascending colon, and low TLRs-expression might contribute to the preferential settlement of indigenous bacteria in MAM. The increase of proliferating epithelial cells is probably induced by the invasion of indigenous bacteria into the intestinal crypts of MAM. KEY WORDS: ascending colon, epithelial proliferation, indigenous bacteria, rat, Toll-like receptor.doi: 10.1292/jvms.13-0036; J. Vet. Med. Sci. 75(7): [939][940][941][942][943][944][945][946][947] 2013 A large number of indigenous bacteria settle in the animal alimentary tract. The bacterial settlement initiates immediately after birth by bacteria derived from mother, and the number and composition of indigenous bacteria are stable in the adult alimentary tract [4]. The number of indigenous bacteria in the chyme increases toward the caudal alimentary tract, that is, 0-10 5 colony forming unit (CFU)/ml in the stomach to jejunum, 10 3 -10 9 CFU/ml in the ileum and 10 10 -10 12 CFU/ml in the colon of human [28], resulting in total 2 kg of indigenous bacteria in the human alimentary tract [11]. From histoplanimetrical investigation along the longitudinal axis of the rat intestine, indigenous bacteria settle on the intestinal superficial epithelium, whose apical surface expresses special sugars throughout the large intestine, and indigenous bacteria also reside in the intestinal crypts in the cecum and proximal 30% of the colon [34,35]. Additionally, numerous indigenous bacteria exist between the mucosal folds in the rat ascending colon [27]. However, the detailed distribution of the settlement sites of indigenous bacteria has not been investigated in the ascending colon.The settlement of indigenous or exogenous bac...

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confidence: 99%

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confidence: 99%

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Histoplanimetrical Study on the Relationship between Invasion of Indigenous Bacteria into Intestinal Crypts and Proliferation of Epithelial Cells in Rat Ascending Colon

Mantani

Takahara

Takeuchi

et al. 2013

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“…These portions are also epithelial cell-shedding sites and accumulation sites of epithelial cells in the late-apoptotic stage in rats [30,33,34]. These accumulation sites of epithelial cells in the lateapoptotic stage also absorb the luminal food-derived antigens and their antibody complexes in the rat small intestine [43,44].…”

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confidence: 94%

Lectin Histochemical Detection of Special Sugars on the Mucosal Surfaces of the Rat Alimentary Tract

Yamamoto

Yokoo

et al. 2010

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ABSTRACT. Surfaces of the most luminal positions of mucosae are fundamental settlement sites of indigenous bacteria throughout the rat alimentary tract. In these positions, also epithelial cell-shedding sites, the special sugar expression in the glycocalyx is very important as it provides possible ligands of bacterial lectins for attachment to epithelial cells. Therefore, the sugar expression in glycocalyx of epithelial cells was lectin-histochemically surveyed using 21 lectins throughout the rat alimentary tract. From the tongue to the nonglandular part of the stomach, -D-Man, -D-Glc and -D-GalNAc were detected on the surface of the keratinized stratified squamous epithelium. In the glandular part of the stomach,(1-4)GlcNAc and bisected triantennary N-glycans were detected on the surface of gastric superficial epithelial cells. From the duodenum to the ileum, (GlcNAc) 2-4 was expressed exclusively on the epithelial cells in the apical portions of the intestinal villi. From the cecum to the rectum,GalNAc) n and NeuNAc were expressed on the intestinal superficial epithelial cells. These results suggest that special sugars are expressed on the most luminal portions of mucosae as exclusive epithelial cell-shedding sites, and that sugar expression differs among the various segments of the alimentary tract. These site differences might reflect differences in resident bacterial species in the rat alimentary tract.KEY WORDS: alimentary tract, indigenous bacteria, lectin histochemistry, rat, sugar expression.

“…The lifespan and migration speed of columnar epithelial cells are crucial factors in the regulation of the settlement of indigenous bacteria in the rat small and large intestine. Therefore, the stable settlement of indigenous bacteria is considered to be maintained by the delicate balance of the proliferation rate of indigenous bacteria over the exfoliation rate of effete apoptotic epithelial cells [20,21]. In addition, the indigenous bacteria closely attached to the epithelial cell membranes are eliminated by physical and chemical elimination by epithelial cells themselves [10,11].…”

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Immunohistochemical and Histoplanimetrical Study on the Spatial Relationship between the Settlement of Indigenous Bacteria and the Secretion of Bactericidal Peptides in Rat Alimentary Tract

Yokoo

Miyata

Udayanga

et al. 2011

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ABSTRACT. To clarify the regulatory mechanism by bactericidal peptides secretion, the secretion of bactericidal peptides was immunohistochemically and histoplanimetrically compared with the degree of Gram-positive/negative bacterial colonization throughout the rat alimentary tract. In the associated exocrine glands from the oral cavity to the stomach, no comparable differences were observed under the changes of development of indigenous bacterial colonies. In the small intestine, immunopositive granules for lysozyme and secretory phospholipase A2 (sPLA2) were markedly decreased, whereas immunopositive vacuoles in the Paneth cells were more increased at sites with hyper-development of indigenous bacterial colonies in the intervillous spaces than at sites with no or less development. No changes in exocrine glands were observed in the large intestine because of the constant existence of large quantities of bacteria. Gram-positive bacterial colonies on the mucosal surfaces were dominant from the oral cavity to the stomach. Gram-negative bacteria were dominant in the large intestine, and the distributions of both Gram-positive and negative bacteria were intermediate in the small intestine. These findings suggest that lysozyme and sPLA2 secreted from the Paneth cells contribute to the regulation of the proliferation of indigenous bacteria in the intervillous spaces of the small intestine, and that the inversion of distributions of Grampositive and -negative bacteria in the alimentary tract might be caused by the secretion of lysozyme and sPLA2 in the small intestine.KEY WORDS: alimentary tract, bactericidal peptides, digestive organs, indigenous bacteria, Paneth cells.