Separation of Intact Intestinal Epithelium from Mesenchyme

Nik, Ali Moussavi; Carlsson, Peter

doi:10.2144/000114055

Cited by 30 publications

(28 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To test whether a 3‐day HFD exposure differentially regulated the gene and protein expression of key metabolic enzymes, we isolated the enterocytes with a novel and very specific method. This method allows for a fast and specific isolation of intestinal epithelial cells without the use of enzymes or EDTA and can be performed in ice cold conditions (Nik and Carlsson, ). We consistently found that 3 and 14 days of HFD feeding reduced the expression of genes related to glucose metabolism.…”

Section: Discussionmentioning

confidence: 99%

Metabolic Adaptation of the Small Intestine to Short- and Medium-Term High-Fat Diet Exposure

et al. 2016

View full text Add to dashboard Cite

The small intestine is the main organ involved in the digestion and absorption of nutrients. It is in an ideal position to sense the availability of energy in the lumen in addition to its absorptive function. Consumption of a high-fat diet (HFD) influences the metabolic characteristics of the small intestine. Therefore, to better understand the metabolic features of the small intestine and their changes in response to dietary fat, we characterized the metabolism of duodenal, jejunal, and hepatic cell lines and assessed the metabolic changes in the enterocytes and the liver after short-term (3 days) or medium-term (14 days) HFD feeding in mice. Experiments with immortalized enterocytes indicated a higher glycolytic capacity in the duodenal cell line compared to the other two cell lines, whereas the jejunal cell line exhibited a high oxidative metabolism. Short-term HFD feeding induced changes in the expression of glucose and lipid metabolism-related genes in the duodenum and the jejunum of mice, but not in the liver. When focusing on fatty acid oxidation both, short- and medium-term HFD feeding induced an upregulation of 3-hydroxy-3-methylglutaryl-coenzyme A, the key enzyme of ketogenesis, at the protein level in the intestinal epithelial cells, but not in the liver. These results suggest that HFD feeding induces an early adaptation of the small intestine rather than the liver in response to a substantial fat load. This highlights the importance of the small intestine in the adaptation of the body to the metabolic changes induced by HFD exposure. J. Cell. Physiol. 232: 167-175, 2017. © 2016 Wiley Periodicals, Inc.

show abstract

Section: Discussionmentioning

confidence: 99%

Metabolic Adaptation of the Small Intestine to Short- and Medium-Term High-Fat Diet Exposure

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Colonic epithelium were isolated using Cell Recovery Solution (Corning) as described (Nik and Carlsson, 2013). Briefly, colons were removed and flushed several times with ice-cold PBS.…”

Section: Star Methodsmentioning

confidence: 99%

IL-22 Upregulates Epithelial Claudin-2 to Drive Diarrhea and Enteric Pathogen Clearance

Tsai

Zhang

et al. 2017

Cell Host & Microbe

209

200

View full text Add to dashboard Cite

SUMMARY Diarrhea is a host response to enteric pathogens, but its impact on pathogenesis remains poorly defined. By infecting mice with the attaching and effacing bacteria Citrobacter rodentium, we defined the mechanisms and contributions of diarrhea and intestinal barrier loss to host defense. Increased permeability occurred within 2 days of infection and coincided with IL-22-dependent upregulation of the epithelial tight junction protein claudin-2. Permeability increases were limited to small molecules, as expected for the paracellular water and Na+ channel formed by claudin-2. Relative to wildtype, claudin-2-deficient mice experienced severe disease, including increased mucosal colonization by C. rodentium, prolonged pathogen shedding, exaggerated cytokine responses, and greater tissue injury. Conversely, transgenic claudin-2 overexpression reduced disease severity. Chemically-induced osmotic diarrhea reduced colitis severity and C. rodentium burden in claudin-2 deficient, but not transgenic, mice, demonstrating that claudin-2-mediated protection is the result of enhanced water efflux. Thus, IL-22-induced claudin-2 upregulation drives diarrhea and pathogen clearance.

show abstract

“…To obtain homogenate from GI tissue, small intestinal epithelium was isolated from mesenchyme following a protocol after Nik and Carlsson70. In brief, small intestine was cut into 4–5 cm long pieces.…”

Section: Methodsmentioning

confidence: 99%

Zinc deficiency and low enterocyte zinc transporter expression in human patients with autism related mutations in SHANK3

Pfaender

Sauer

Hagmeyer

et al. 2017

Sci Rep

View full text Add to dashboard Cite

Phelan McDermid Syndrome (PMDS) is a genetic disorder characterized by features of Autism spectrum disorders. Similar to reports of Zn deficiency in autistic children, we have previously reported high incidence of Zn deficiency in PMDS. However, the underlying mechanisms are currently not well understood. Here, using inductively coupled plasma mass-spectrometry to measure the concentration of Zinc (Zn) and Copper (Cu) in hair samples from individuals with PMDS with 22q13.3 deletion including SHANK3 (SH3 and multiple ankyrin repeat domains 3), we report a high rate of abnormally low Zn/Cu ratios. To investigate possible underlying mechanisms, we generated enterocytes from PMDS patient-derived induced pluripotent stem cells and used Caco-2 cells with knockdown of SHANK3. We detected decreased expression of Zn uptake transporters ZIP2 and ZIP4 on mRNA and protein level correlating with SHANK3 expression levels, and found reduced levels of ZIP4 protein co-localizing with SHANK3 at the plasma membrane. We demonstrated that especially ZIP4 exists in a complex with SHANK3. Furthermore, we performed immunohistochemistry on gut sections from Shank3αβ knockout mice and confirmed a link between enterocytic SHANK3, ZIP2 and ZIP4. We conclude that apart from its well-known role in the CNS, SHANK3 might play a specific role in the GI tract.

show abstract

Separation of Intact Intestinal Epithelium from Mesenchyme

Cited by 30 publications

References 16 publications

Metabolic Adaptation of the Small Intestine to Short- and Medium-Term High-Fat Diet Exposure

Metabolic Adaptation of the Small Intestine to Short- and Medium-Term High-Fat Diet Exposure

IL-22 Upregulates Epithelial Claudin-2 to Drive Diarrhea and Enteric Pathogen Clearance

Zinc deficiency and low enterocyte zinc transporter expression in human patients with autism related mutations in SHANK3

Contact Info

Product

Resources

About