Basic–alimentary tractInterferon-γ Inhibits Intestinal Restitution by Preventing Gap Junction Communication Between Enterocytes
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Cell Culture, Transfection, and Treatment
Cultured intestinal cell lines (small intestine: IEC-6 and large intestine: HT-29, T84, and CaCO-2) were obtained from the American Type Culture Collection (Manassas, VA) and maintained as described.19, 20, 21 Where indicated, cells were treated with IFN-gamma (IFN, Sigma-Aldrich Corp., St. Louis, MO, 1000 IU/mL for 1–14 hours, 37°C), or the following phosphatase inhibitors 20 minutes to 1 hour prior to IFN treatment: calyculin A (2 μmol/L), okadaic acid (100 nmol/L, Calbiochem, San Diego, CA),
Enterocytes Express Functional Gap Junctions
We first sought to determine whether enterocytes express the gap junction protein Cx43 (Cx43), and whether gap junctions are capable of exchanging molecules between adjacent enterocytes. As shown in Figure 1, Cx43, which is the most widely expressed connexin isoform, was detected in several cultured enterocyte cell lines, including the small intestinal epithelial cell line IEC-6, the colonic epithelial lines CaCO-2, HT-29, and T84, and in mucosal scrapings from the terminal ilea of
Discussion
NEC is characterized by damage to the mucosal barrier and a profound inhibition in the rate and extent of enterocyte migration.9 Repair of the damaged intestine occurs through the process of intestinal restitution, in which healthy enterocytes adjacent to the site of injury migrate to areas of mucosal disruption to reconstitute an intact barrier.37 Despite excellent work from a variety of authors studying the regulation of enterocyte migration,38, 39 few studies have distinguished the
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D. J. H. was supported by 1R01-GM078238-01 from the National Institutes of Health and the State of Pennsylvania Tobacco Settlement Fund. C. L. L. was supported in part by the Loan Repayment Program for Pediatric Research of the National Institutes of Health.
There are no conflicts of interest to disclose with respect to this manuscript.