Positional specificity of defensin gene expression reveals Paneth cell heterogeneity in mouse small intestine

1996 ◽  
Vol 271 (1) ◽  
pp. G68-G74 ◽  
Author(s):  
D. Darmoul ◽  
A. J. Ouellette
Keyword(s):  
Gene Expression ◽  
Small Intestine ◽  
Small Bowel ◽  
Paneth Cell ◽  
Paneth Cells ◽  
Northern Blot Hybridization ◽  
Positional Specificity ◽  
Defensin Gene ◽  
Proximal Small Bowel ◽  
Mouse Small Intestine

Cryptdins are antimicrobial peptides of the defensin family that are expressed specifically by Paneth cells in small intestinal crypts (M.E. Selsted, S.I. Miller, A.H. Henschen, and A.J. Ouellette. J. Cell Biol. 118: 929-936, 1992), and at least 17 cryptdin isoforms have been reported in mouse small intestine (A.J. Ouellette, M.M. Hsieh, M.T. Nosek, D.F. Cano-Gauci, K.M. Huttner, R.N. Buick, and M.E. Selsted. Infect. Immun. 62: 5040-5047, 1994). Analysis of cryptdin gene expression in adult mouse small bowel revealed that the cryptdin-4 isoform is differentially expressed along the proximal-to-distal intestinal axis. By peptide-specific reverse transcriptase-polymerase chain reaction-based assays, cryptdin-4 mRNA was found to be absent from the proximal small bowel, increasing to maximal levels in the ileum. In contrast, intestinal content of cryptdin-1 and -5 mRNAs was equivalent in duodenum, jejunum, and ileum, and Northern blot hybridization experiments were consistent with both sets of data. Similarly, individual crypts isolated from duodenum contain cryptdin-1 mRNA but not cryptdin-4 mRNA. Taken together, the results show that Paneth cells are heterogeneous, depending on their position along the longitudinal axis of the small bowel. The positional specificity of defensin gene expression suggests that cryptdins may be useful markers for investigating the establishment and maintenance of this epithelial lineage in the mouse small intestine.

scholarly journals Methyl donor deficiency affects small-intestinal differentiation and barrier function in rats

2012 ◽  
Vol 109 (4) ◽  
pp. 667-677 ◽  
Author(s):  
Aude Bressenot ◽  
Shabnam Pooya ◽  
Carine Bossenmeyer-Pourie ◽  
Guillaume Gauchotte ◽  
Adeline Germain ◽  
...  
Keyword(s):  
Small Intestine ◽  
Small Bowel ◽  
Barrier Function ◽  
Paneth Cell ◽  
Cell Number ◽  
Nuclear Antigen ◽  
Methyl Donors ◽  
Distal Small Bowel ◽  
Proximal Small Bowel ◽  
Enterocyte Differentiation

Dietary methyl donors and their genetic determinants are associated with Crohn's disease risk. We investigated whether a methyl-deficient diet (MDD) may affect development and functions of the small intestine in rat pups from dams subjected to the MDD during gestation and lactation. At 1 month before pregnancy, adult females were fed with either a standard food or a diet without vitamin B12, folate and choline. A global wall hypotrophy was observed in the distal small bowel (MDD animals 0·30 mm v. controls 0·58 mm; P< 0·001) with increased crypt apoptosis (3·37 v. 0·4 %; P< 0·001), loss of enterocyte differentiation in the villus and a reduction in intestinal alkaline phosphatase production. Cleaved caspase-3 immunostaining (MDD animals 3·37 % v. controls 0·4 %, P< 0·001) and the Apostain labelling index showed increased crypt apoptosis (3·5 v. 1·4 %; P= 0·018). Decreased proliferation was observed in crypts of the proximal small bowel with a reduced number of minichromosome maintenance 6 (MDD animals 52·83 % v. controls 83·17 %; P= 0·048) and proliferating cell nuclear antigen-positive cells (46·25 v. 59 %; P= 0·05). This lack of enterocyte differentiation in the distal small bowel was associated with an impaired expression of β-catenin and a decreased β-catenin–E-cadherin interaction. The MDD affected the intestinal barrier in the proximal small bowel by decreasing Paneth cell number after immunostaining for lysosyme (MDD animals 8·66 % v. controls 21·66 %) and by reducing goblet cell number and mucus production after immunostaining for mucin-2 (crypts 8·66 v. 15·33 %; villus 7 v. 17 %). The MDD has dual effects on the small intestine by producing dramatic effects on enterocyte differentiation and barrier function in rats.

Maximal lymphatic triglyceride transport rate from the rat small intestine

1982 ◽  
Vol 242 (4) ◽  
pp. G408-G415 ◽  
Author(s):  
P. Tso ◽  
K. L. Buch ◽  
J. A. Balint ◽  
J. B. Rodgers
Keyword(s):  
Small Intestine ◽  
Small Bowel ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Transport Rate ◽  
Lymphatic Transport ◽  
Onset Of Action ◽  
Triglyceride Fatty Acid ◽  
Proximal Half ◽  
Proximal Small Bowel

In previous studies, we demonstrated that the hydrophobic surfactant Pluronic L-81 blocks lymphatic triglyceride transport from the small intestine and leads to accumulation of triglyceride in the mucosa. The onset of action of Pluronic L-81 is rapid and quickly reversed once its administration is discontinued. We have taken advantage of these effects of Pluronic L-81 on intestinal lipid transport in order to determine the apparent maximal triglyceride transport capacity of the proximal half of the rat small bowel using lymph fistula rats infused intraduodenally with a phosphate-buffered, taurocholate-stabilized emulsion containing 40 mumol [3H]triolein and 0.5 mg Pluronic L-81 at 3 ml/h for 8 h to load the proximal small bowel with lipid. Studies were done in one group of rats in order to be certain that only the proximal half of the small bowel contained 3H-lipid after this period of infusion. In other rats treated similarly, the 8 h of lipid-Pluronic L-81 infusion were followed by infusion of 3 ml/h of 0.15 M salt solution for 5 h. Lymphatic transport of lipid was determined throughout the entire period of infusion. During lipid-Pluronic L-81 infusion, transport of 3H-triglyceride fatty acid into lymph was only 22-27 mumol/h but rose steadily after substitution of saline and reached a maximal transport rate of 109 +/- 6.2 mumol/h (means +/- SE) after 3.5 h. During this 3.5-h period, the amount of 3H-lipid in the proximal mucosa declined from 530 to 263 mumol. While Pluronic L-81 was infused, only very low-density-lipoprotein-sized particles were seen in lymph by electron microscopy, whereas, at the peak of triglyceride transport during saline infusion, chylomicrons of up to 6,000 A were observed in lymph.

scholarly journals Bacterial Overgrowth in the Cystic Fibrosis Transmembrane Conductance Regulator Null Mouse Small Intestine

2004 ◽  
Vol 72 (10) ◽  
pp. 6040-6049 ◽  
Author(s):  
Oxana Norkina ◽  
Tim G. Burnett ◽  
Robert C. De Lisle
Keyword(s):  
Cystic Fibrosis ◽  
Small Intestine ◽  
Intestinal Inflammation ◽  
Bacterial Load ◽  
Paneth Cell ◽  
Bacterial Overgrowth ◽  
Intestinal Lumen ◽  
Cytokine Signaling ◽  
Null Mouse ◽  
Mouse Small Intestine

ABSTRACT We recently reported the inflammation of the cystic fibrosis (CF) mouse small intestine, and we hypothesized bacterial overgrowth as a possible cause. Quantitative PCR of bacterial 16S genomic DNA in the CF mouse small intestine revealed an increase of greater than 40-fold compared to controls. Sequencing of 16S PCR products and Gram staining showed that the majority of bacteria in the CF mouse intestine were gram negative. Bacteria were observed to colonize the mucus that accumulates in the intestinal lumen of mice with CF. Impaired Paneth cell defenses were suggested by observation of partially dispersed Paneth granules in the mucus plugs of CF mouse intestinal crypts, and this mucus was strongly immunoreactive for Paneth cell bactericidal products. The role of bacterial overgrowth in intestinal inflammation in CF was tested by treating mice with oral antibiotics (ciprofloxacin and metronidazole) for 3 weeks, which reduced bacterial load in the CF mouse small intestine over 400-fold. Antibiotic treatment decreased the expression of the inflammation-related genes mast cell protease 2, leucine-rich α2 glycoprotein/leucine-rich high endothelial venule glycoprotein, suppressor of cytokine signaling 3, hematopoietic cell transcript 1, and resistin-like molecule β/found in inflammatory zone 2, all of which were no longer expressed at levels significantly different from control levels. The reduction of intestinal bacteria also significantly improved the growth of CF mice but had no effect on the growth of wild-type mice. These data suggest that bacterial overgrowth in the CF mouse small intestine has a role in inflammation and contributes to the failure to thrive in this mouse model of CF.

Effects of bile and pancreatic secretions on intestinal mucosa after proximal small bowel resection in rats

1980 ◽  
Vol 58 (9) ◽  
pp. 1117-1123 ◽  
Author(s):  
Monique D. Gélinas ◽  
Claude L. Morin
Keyword(s):  
Small Intestine ◽  
Small Bowel ◽  
Pancreatic Juice ◽  
Bowel Resection ◽  
Intestinal Resection ◽  
Small Bowel Resection ◽  
Sprague Dawley ◽  
Ileal Mucosa ◽  
Distal Small Intestine ◽  
Proximal Small Bowel

After proximal small bowel resection the remaining small intestine undergoes adaptive hyperplasia. In the present study, the relative contributions of bile and (or) pancreatic juice to adaptive intestinal hyperplasia following proximal resection was studied. Using male Sprague–Dawley rats a 50% proximal intestinal resection was done starting 10 cm distal to the beginning of the jejunum. The animals were also subjected to diversion of bile and (or) pancreatic secretions to the distal ileum at 18 cm proximal to the ileocecal junction. After 8 days gut and mucosal weights, mucosal proteins, and DNA were measured in the duodenojejunum (gut segment proximal to the resection anastomosis) and in the ileum (first half of the small bowel segment distal to the diversion site). The results indicate that (1) in rats fed either chow (Purina rat chow) or a chemically defined diet diversion of pancreaticobiliary secretions to the ileum significantly stimulated ileal mucosa growth whereas no changes were observed in the duodenojejunum, (2) in rats fed a chemically defined diet neither bile nor pancreatic juice affected ileal mucosa when separately diverted to the ileum, and (3) pancreatic juice draining into the duodenum while bile was diverted to the ileum induced hypoplastic changes in the duodenojejunum. The present study suggests that following jejunectomy the regulation of mucosal growth by pancreatic and bile secretions is different in the proximal and distal small intestine. Pancreaticobiliary secretions are trophic for the ileum. However, in the proximal gut bile offers protection against a direct or indirect catabolic action of pancreatic juice.

scholarly journals Activation of Paneth Cell α-Defensins in Mouse Small Intestine

2001 ◽  
Vol 277 (7) ◽  
pp. 5219-5228 ◽  
Author(s):  
Tokiyoshi Ayabe ◽  
Donald P. Satchell ◽  
Patrizia Pesendorfer ◽  
Hiroki Tanabe ◽  
Carole L. Wilson ◽  
...  
Keyword(s):  
Small Intestine ◽  
Paneth Cell ◽  
Mouse Small Intestine

T1248 Regulated Roles of Paneth Cell α-Defensin in Mouse Small Intestine

2008 ◽  
Vol 134 (4) ◽  
pp. A-515
Author(s):  
Rie Fukaya ◽  
Naoki Sakai ◽  
Tokiyoshi Ayabe
Keyword(s):  
Small Intestine ◽  
Paneth Cell ◽  
Mouse Small Intestine

Inflammation of the cystic fibrosis mouse small intestine

2004 ◽  
Vol 286 (6) ◽  
pp. G1032-G1041 ◽  
Author(s):  
Oxana Norkina ◽  
Simran Kaur ◽  
Donna Ziemer ◽  
Robert C. De Lisle
Keyword(s):  
Gene Expression ◽  
Cystic Fibrosis ◽  
Small Intestine ◽  
Immune System ◽  
Mast Cells ◽  
Innate Immune System ◽  
Innate Immune ◽  
Serum Amyloid ◽  
Mouse Small Intestine ◽  
Cytochrome P 450

The CFTR null mouse [cystic fibrosis (CF) mouse] has a severe intestinal phenotype that serves as a model for CF-related growth deficiency, meconium ileus, and distal intestinal obstructive syndrome. DNA microarray analysis was used to investigate gene expression in the CF mouse small intestine. Sixty-one genes exhibited a statistically significant twofold or greater increase in expression, and 98 genes were downregulated twofold or greater. Of the upregulated genes, most were associated with inflammation and included markers for cells of the innate immune system (mast cells and neutrophils) and for acute-phase genes (serum amyloid A and complement factors). The downregulated genes include 10 cytochrome P-450 genes; several are involved in lipid metabolism, and several are involved in various transport processes. Confirmation by quantitative RT-PCR showed gene expression was significantly increased for mast cell protease 2 (27-fold), hematopoietic cell transcript 1 (17-fold), serum amyloid A3 (2.9-fold), suppressor of cytokine signaling 3 (2.0-fold), leucine-rich α2-glycoprotein (21-fold), resistin-like molecule-β (49-fold), and Muclin (2.5-fold) and was significantly decreased for cytochrome P-450 4a10 (28-fold) and cubilin (114-fold). Immune cell infiltration was confirmed histologically by staining for mast cells and neutrophils. These data demonstrate that the CF intestine exhibits an inflammatory state with upregulation of components of the innate immune system.

scholarly journals Immunohistochemical characterization of a crypt cell-specific plasma membrane protein in rat small intestine epithelium using a monoclonal antibody.

1991 ◽  
Vol 39 (7) ◽  
pp. 973-979 ◽  
Author(s):  
H Schiechl
Keyword(s):  
Monoclonal Antibody ◽  
Small Intestine ◽  
Protein Pattern ◽  
Post Partum ◽  
Paneth Cell ◽  
Paneth Cells ◽  
Crypt Cell ◽  
Fluorescence Labeling ◽  
Cell Protein ◽  
Undifferentiated Cells

Proteins of the basolateral membrane (BLM) of small intestine epithelial cells of adult rats, in the MW ranges of 50-65 KD, 85-100 KD, and over 100 KD, were obtained as follows. After isolation of the BLM and subsequent SDS-PAGE and transblotting of the proteins on nitrocellulose sheets, the bands in these MW ranges were cut out of the nitrocellulose sheet and extracted. Balb/C mice were immunized with these protein fractions and a monoclonal antibody (MAb) was then produced. MAb SI/CC1 obtained via immunization with the 50-65 KD protein fraction shows specificity for the crypt epithelium of the small intestine. It can be used to characterize, by light and electron microscopic immunohistochemical methods, a crypt cell protein (SI/CC1-Ag) with a very specific localization. Fluorescence labeling shows that the SI/CC1-Ag can be found only in the epithelium of small intestine crypts (except for the granules in eosinophilic granulocytes). The epithelium of the colon, as well as the epithelia of other organs, could not be labeled. In the small intestine crypts, SI/CC1-Ag is found only in the Paneth cells located in the basal crypt section, and in the undifferentiated cells in the middle crypt section; it is lacking in the cells of the upper crypt section. Gold labeling shows that SI/CC1-Ag in the undifferentiated cells is localized exclusively in the basolateral PM domain. On the Paneth cells, the content of the secretory granules is labeled, along with the basolateral PM domain; the labeling sometimes present on their luminal part is probably due to passively absorbed secretion from these cells. The SI/CC1-Ag in the BLM of undifferentiated and Paneth cells is found only on Days 21-23 post partum, whereas the Paneth cell granules could be labeled as early as the Day 16 post partum. With immunodetection with SI/CC1, one band at about 55 KD is specifically labeled in the protein pattern of the isolated small intestine cell BLM. In the protein pattern of the isolated crypt cells two bands were labeled, again one at 55 KD and one at about 120 KD. These findings indicate that SI/CC1-Ag is a 55 KD protein that appears on Days 21-23 post partum in the BLM of undifferentiated cells and of Paneth cells.

Changes in goblet cell homeostasis, mucin structure and gene expression in the mouse small intestine during rotavirus infection

2001 ◽  
Vol 120 (5) ◽  
pp. A125-A125
Author(s):  
J BOSHUIZEN ◽  
J REIMERINK ◽  
V HAM ◽  
R BINDA ◽  
M KOOPMANS ◽  
...  
Keyword(s):  
Gene Expression ◽  
Small Intestine ◽  
Goblet Cell ◽  
Cell Homeostasis ◽  
Rotavirus Infection ◽  
Mouse Small Intestine
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