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BMC Gastroenterol. 2015 Apr 15;15:47. doi: 10.1186/s12876-015-0275-5.

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

Author information

1
Department of Clinical and Translational Sciences, Joan C. Edwards School of Medicine, Marshall University, 1600 Hal Greer Blvd., Huntington, WV, 25701, USA. singhs@marshall.edu.
2
Department of Clinical and Translational Sciences, Joan C. Edwards School of Medicine, Marshall University, 1600 Hal Greer Blvd., Huntington, WV, 25701, USA. arthursu@marshall.edu.
3
Department of Biology, LeMoyne-Owen College, Memphis, TN, 38126, USA. Jamil-Talukder@loc.edu.
4
Department of Clinical and Translational Sciences, Joan C. Edwards School of Medicine, Marshall University, 1600 Hal Greer Blvd., Huntington, WV, 25701, USA. palaniappan@marshall.edu.
5
Boston University School of Medicine, Boston, MA, 02118, USA. coons@bu.edu.
6
Department of Clinical and Translational Sciences, Joan C. Edwards School of Medicine, Marshall University, 1600 Hal Greer Blvd., Huntington, WV, 25701, USA. sundaramu@marshall.edu.

Abstract

BACKGROUND:

In 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.

METHODS:

Chronic 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 (3)H-Glutamine. Western blot analyses were done using B0AT1 and SN2 specific antibodies.

RESULTS:

In 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.

CONCLUSION:

In 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.

PMID:
25884559
PMCID:
PMC4405831
DOI:
10.1186/s12876-015-0275-5
[Indexed for MEDLINE]
Free PMC Article

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