Novel ex Vivo Experimental Setup to Assay the Vectorial Transepithelial Enteroendocrine Secretions of Different Intestinal Segments

J Agric Food Chem. 2018 Nov 7;66(44):11622-11629. doi: 10.1021/acs.jafc.8b03046. Epub 2018 Oct 26.

Abstract

The enteroendocrine system coordinates gastrointestinal (GI) tract functionality and the whole organism. However, the scarcity of enteroendocrine cells and their scattered distribution make them difficult to study. Here, we glued segments of the GI wall of pigs to a silicon tube, keeping the apical and the basolateral sides separate. The fact that there was less than 1% of 70-kDa fluorescein isothiocyanate (FITC)-dextran on the basolateral side proved that the gluing was efficient. Since the lactate dehydrogenase leakage at basolateral side was lower than 0.1% (1.40 ± 0.17 nKatals) it proved that the tissue was viable. The intestinal barrier function was maintained as it is in segments mounted in Ussing chambers (the amount of Lucifer Yellow crossing it, was similar between them; respectively, % LY, 0.48 ± 0.13; 0.52 ± 0.09; p > 0.05). Finally, apical treatments with two different extract produced differential basolateral enterohormone secretions (basolateral PYY secretion vs control; animal extract, 0.35 ± 0.16; plant extract, 2.5 ± 0.74; p < 0.05). In conclusion, we report an ex vivo system called "Ap-to-Bas" for assaying vectorial transepithelial processes that makes it possible to work with several samples at the same time. It is an optimal device for enterohormone studies in the intestine.

Keywords: enteroendocrine system; ex vivo model; food bioactives; gut hormone secretion; transepithelial activity.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Biological Assay / instrumentation
  • Biological Assay / methods*
  • Enteroendocrine Cells / metabolism*
  • Female
  • In Vitro Techniques / instrumentation
  • In Vitro Techniques / methods*
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / metabolism
  • Intestines / cytology*
  • Peptide YY / metabolism
  • Swine

Substances

  • Peptide YY