Development of Small Intestinal Preparation with Intact Epithelial Polarity for Patch-Clamp Study

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The epithelial cells of the mammalian small intestine partakes in both secretion of Cl-and absorption of Na+as well as organic solutes.It is generally thought that the intestinal secretion is carried out by the crypt epithelial cell whereas the absorption is by the villus epithelial cell. However, little direct level evidence has been provided for the hypothesis. To directly examine the hypothesis at the single cell level, we had started experiments using single villus epithelial cells isolated from the guinea pig small intestine.The brush-border structure of singly isolated enterocytes, however, became rapidly lost, and therefore no glucose-dependent ionic currents could be recorded from them by the patch-clamp technique. The present study is aimed at developing the method to prepare the villus and crypt specimen with the intact epithelial polarity from the small intestine of the adult guinea pig. The preparations could be separately obtained by isolating the epithelia after vibration in the EDTA-containing saline solution and by slicing the tissue. By applying the patch-clamp technique, numbers of ionic currents could be recorded from these isolated epithelia, especially glucose-dependent ionic currents from isolated villi and cyclic AMP-activated (CFTR-typed)Cl-channel currents from isolated crypts.