scholarly journals A low-temperature method for the isolation of small-intestinal epithelium along the crypt-villus axis

1991 ◽  
Vol 280 (2) ◽  
pp. 331-334 ◽  
Author(s):  
N Flint ◽  
F L Cove ◽  
G S Evans

A variety of enzymic and non-enzymic methods to isolate epithelium from the small intestine have been previously published. Sequential fractionation of cells from the villus to the crypt has been reported in some of these papers, which allows the comparative study of terminally differentiated and proliferative cell phenotypes. However, these methods often involve the incubation of tissues at 37 degrees C, which may affect the structural and biochemical integrity of the cells. We have developed a rapid low-temperature (4 degrees C) method for isolating purified populations of crypt and villus cells from mouse and rat intestines. The fractionated cells have been partially characterized, and the potential value of the procedure has been indicated by the ability to analyse the comparative protein and mRNA expression along the crypt-villus axis.

1997 ◽  
Vol 110 (11) ◽  
pp. 1317-1324 ◽  
Author(s):  
I. Duluc ◽  
O. Lorentz ◽  
C. Fritsch ◽  
C. Leberquier ◽  
M. Kedinger ◽  
...  

In segmented organs, homeobox genes are involved in axial patterning and cell identity. Much less is known about their role in non-segmented endoderm derivatives such as the digestive epithelium. Using a xenograft model of fetal intestinal anlagen implanted under the skin of nude mice, we have investigated whether the expression of five homeobox genes (HoxA-4, HoxA-9, HoxC-8, Cdx-1 and Cdx-2) is modified when intestinal epithelium undergoes normal development or displays heterodifferentiation in association with heterotopic mesenchyme. In homotypic associations of fetal endoderm and mesenchyme that recapitulate normal development, the overall pattern of homeobox gene expression was maintained: HoxA-9 and HoxC-8 were the highest in the colon and ileum, respectively, and HoxA-4 was expressed all along the intestine; Cdx-1 and Cdx-2 exhibited an increasing gradient of expression from small intestine to colon. Yet, grafting per se caused a faint upregulation of HoxA-9 and HoxC-8 in small intestinal regions in which these genes are not normally expressed, while the endoderm-mesenchyme dissociation-association step provoked a decay of Cdx-1 in the colon. In heterotopic associations of colonic endoderm with small intestinal mesenchyme, the colonic epithelium exhibited heterodifferentiation to a small intestinal-like phenotype. In this case, we observed a decay of HoxA-9 expression and an upregulation of HoxC-8. Additionally, heterodifferentiation of the colonic epithelium was accompanied by a downregulation of Cdx-1 and Cdx-2 to a level similar to that found in the normal small intestine. To demonstrate that mesenchyme-derived cells can influence Cdx-1 and Cdx-2 expression in the bowel epithelium, fetal jejunal endoderm was associated with intestinal fibroblastic cell lines that either support small intestinal-like or colonic-like morphogenesis. A lower expression of both homeobox genes was shown in grafts presenting the small intestinal phenotype than in those showing glandular colonic-like differentiation. Taken together, these results suggest that homeobox genes participate in the control of the positional information and/or cell differentiation in the intestinal epithelium. They also indicate that the level of Cdx-1 and Cdx-2 homeobox gene expression is influenced by epithelial-mesenchymal cell interactions in the intestinal mucosa.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Paul Cray ◽  
Breanna J. Sheahan ◽  
Jocsa E. Cortes ◽  
Christopher M. Dekaney

AbstractEnteric bacteria and/or their products are necessary for doxorubicin (DXR)-induced small intestine mucosal damage. While DXR does not induce gross loss of epithelium, others have shown elevated serum endotoxin after DXR administration. However, the mechanism of movement is unknown. We hypothesized that DXR treatment resulted in increased paracellular translocation of bacteria or bacterial products through the small intestinal epithelium. We measured permeability after DXR administration using transepithelial resistance and macromolecular flux and assessed tight junctional gene expression and protein localization both in vitro using T84 cells and ex vivo using murine jejunum. DXR treatment increased flux of 4 kDa dextrans in mouse jejenum, but increased flux of 4, 10 and 20 kDa dextrans in T84 cells. Following DXR, we observed increased permeability, both in vitro and ex vivo, independent of bacteria. DXR induced increased expression of Cldn2 and Cldn4 in murine small intestine but increased only CLDN2 expression in T84 cells. DXR treatment induced disorganization of tight junctional proteins. We conclude that DXR increases paracellular transit of small macromolecules, including bacterial products, through the epithelium, by altering expression of tight junctional components and dynamic loosening of cellular tight junctions.


2021 ◽  
pp. 1422-1428
Author(s):  
Daisuke Inoue ◽  
Shoji Oura ◽  
Tomoya Takami ◽  
Shinichiro Makimoto

A 69-year-old man with abdominal distention was referred to our hospital. The patient had undergone laparoscopic surgery for his Borrmann type 2 rectal cancer 2 years before. In addition to the re-elevation of serum CEA and CA19-9 levels, computed tomography (CT) showed intestinal dilatation, and positron emission CT showed a presumed tumor with abnormal fluorodeoxyglucose accumulation in the small intestine. We judged the small intestinal dilatation was highly due to the solitary recurrent peritoneal dissemination of rectal cancer and performed laparoscopic evaluation of the abdominal cavity followed by laparoscopic resection of the affected small intestine. The small intestinal tumor resembled the rectal cancer both on macroscopical and microscopical findings, that is, Borrmann type 2 phenotype and adenocarcinoma that was well differentiated in the part that protruded into the small intestinal lumen and whose degree of differentiation gradually decreased toward the serosa. In addition, abrupt disruption of the normal small intestinal epithelium and the lymphocytic aggregation, presumed tumor-infiltrating lymphocytes, just between the tumor and the small intestinal epithelium highly suggested the tumor originating from the colon cancer. The patient recovered uneventfully with marked decrease in tumor marker levels 1 month after the operation but did not receive colon cancer-oriented chemotherapy as adjuvant therapy for his financial reasons. Oncologists should note this type of recurrence to properly treat the patients with recurrent colorectal cancer.


2002 ◽  
Vol 2002 ◽  
pp. 104-104
Author(s):  
J. A. N. Mills ◽  
E. Kebreab ◽  
L. A. Crompton ◽  
J. Dijkstra ◽  
J. France

The high contribution of postruminal starch digestion (>50%) to total tract starch digestion on certain energy dense diets (Mills et al. 1999) demands that limitations to small intestinal starch digestion are identified. Therefore, a dynamic mechanistic model of the small intestine was constructed and evaluated against published experimental data for abomasal carbohydrate infusions in the dairy cow. The mechanistic structure of the model allowed the current biological knowledge to be integrated into a system capable of identifying restrictions to dietary energy recovery from postruminal starch delivery.


2001 ◽  
Vol 280 (3) ◽  
pp. G368-G380 ◽  
Author(s):  
Einar Husebye ◽  
Per M. Hellström ◽  
Frank Sundler ◽  
Jie Chen ◽  
Tore Midtvedt

The effect of an intestinal microflora consisting of selected microbial species on myoelectric activity of small intestine was studied using germ-free rat models, with recording before and after specific intestinal colonization, in the unanesthetized state. Intestinal transit, neuropeptides in blood (RIA), and neuromessengers in the intestinal wall were determined. Clostridium tabificum vp 04 promoted regular spike burst activity, shown by a reduction of the migrating myoelectric complex (MMC) period from 30.5 ± 3.9 min in the germ-free state to 21.2 ± 0.14 min ( P < 0.01). Lactobacillus acidophilus A10 and Bifidobacterium bifidum B11 reduced the MMC period from 27.9 ± 4.5 to 21.5 ± 2.1 min ( P < 0.02) and accelerated small intestinal transit ( P < 0.05). Micrococcus luteus showed an inhibitory effect, with an MMC period of 35.9 ± 9.3 min compared with 27.7 ± 6.3 min in germ-free rats ( P < 0.01). Inhibition was indicated also for Escherichia coli X7gnotobiotic rats. No consistent changes in slow wave frequency were observed. The concentration of neuropeptide Y in blood decreased after introduction of conventional intestinal microflora, suggesting reduced inhibitory control. Intestinal bacteria promote or suppress the initiation and aboral migration of the MMC depending on the species involved. Bacteria with primitive fermenting metabolism (anaerobes) emerge as important promoters of regular spike burst activity in small intestine.


2017 ◽  
Vol 85 (5) ◽  
pp. AB316
Author(s):  
Ryoichi Sawada ◽  
Ryosuke Miyazaki ◽  
Ayako Ishii ◽  
Yusuke Nagata ◽  
Makio Ogawa ◽  
...  

2016 ◽  
Vol 19 (1) ◽  
pp. 48-56 ◽  
Author(s):  
Maria C Jugan ◽  
John R August

Objectives The aim of the study was to evaluate ultrasonographic changes in the small intestine of cats with clinical signs of gastrointestinal disease and low or low–normal serum cobalamin concentrations. Methods Records for client-owned cats presenting to the small animal hospital with signs of gastrointestinal disease and in which serum cobalamin concentrations were measured from 2000–2013 were reviewed. Inclusion criteria were cobalamin concentrations <500 ng/l, abdominal ultrasound within 1 month of cobalamin testing and definitive diagnosis. Results Of 751 serum cobalamin measurements, hypocobalaminemia or low–normal cobalamin was identified in 270 cats, abdominal ultrasound was performed in 207 of those cats and a diagnosis was available for 75 of them. Small intestinal ultrasound changes were detected in 49/75 (65%) cats. Abnormalities included thickening, loss of wall layer definition, echogenicity alterations and discrete masses. Serum cobalamin concentrations <500 ng/l were observed with diagnoses of inflammatory disease, neoplasia, infectious disease and normal histopathology. Cobalamin concentration was significantly lower in cats with lymphoma or inflammatory bowel disease compared with other gastrointestinal neoplasia ( P = 0.031). No difference was found between cobalamin concentration and the presence of ultrasound abnormalities, specific ultrasound changes or albumin concentration. Conclusions and relevance One-third of symptomatic cats with hypocobalaminemia or low–normal cobalamin concentrations may have an ultrasonographically normal small intestine. For the majority of cats in this study, histopathologic abnormalities were observed in the small intestine, regardless of ultrasound changes. These findings suggest gastrointestinal disease should not be excluded based on low–normal cobalamin concentrations, even with a concurrent normal ultrasound examination. Additional studies are needed in cats with low–normal serum cobalamin concentrations, as a definitive diagnosis was not pursued consistently in those cats. However, data from this study suggest that careful monitoring, histopathologic evaluation and future cobalamin supplementation may be warranted.


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