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Updated: May 27, 2026

Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D
Published on: March 16, 2017
In vitro studies on intestinal calcium and phosphate transport in horses
Anja Cehak1, Mirja R Wilkens, Maria Guschlbauer
1Department of Physiology, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173 Hannover, Germany. anja.cehak@tiho-hannover.de
Abstract:
Transepithelial transport mechanisms play a key role in regulating the absorption and secretion of calcium (Ca(2+)) and inorganic phosphate (P(i)) in the gastrointestinal tract. Although intestinal disorders with imbalances in macromineral homeostasis are frequently observed in horses, available data on intestinal Ca(2+) and P(i) transport are limited. The aim of the present study was to characterize the intestinal Ca(2+) and P(i) transport functionally by using the in vitro radioisotope tracer technique with Ussing chambers and to identify components involved in Ca(2+) transport at both mRNA and protein level. Among the different intestinal segments, the duodenum showed significant and highest active Ca(2+) absorption. The findings from RT-PCR and Western blot analysis suggest that the epithelial Ca(2+) channel TRPV6, the cytosolic calcium binding protein calbindin-D(9K) and the plasma membrane calcium ATPase PMCA may be involved in active transcellular Ca(2+) transport. Regarding the P(i) transport, the results indicate significant active P(i) secretion in the jejunum, but the contributing mechanisms remain unclear. A significant inhibiting effect of ouabain as an antagonist of the basolateral Na(+)/K(+)-ATPase on the serosal-to-mucosal P(i) transport suggests a pivotal role of Na(+) in jejunal P(i) transport in the horse.
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