TY - JOUR
T1 - Interferon-γ Inhibits Intestinal Restitution by Preventing Gap Junction Communication Between Enterocytes
AU - Leaphart, Cynthia L.
AU - Qureshi, Faisal
AU - Cetin, Selma
AU - Li, Jun
AU - Dubowski, Theresa
AU - Batey, Catherine
AU - Beer-Stolz, Donna
AU - Guo, Fengli
AU - Murray, Sandra A.
AU - Hackam, David J.
N1 - Funding Information:
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.
PY - 2007/6
Y1 - 2007/6
N2 - Background & Aims: Necrotizing enterocolitis (NEC) is characterized by interferon-gamma (IFN-γ) release and inadequate intestinal restitution. Because enterocytes migrate together, mucosal healing may require interenterocyte communication via connexin 43-mediated gap junctions. We hypothesize that enterocyte migration requires interenterocyte communication, that IFN impairs migration by impairing connexin 43, and that impaired healing during NEC is associated with reduced gap junctions. Methods: NEC was induced in Swiss-Webster or IFN-/- mice, and restitution was determined in the presence of the gap junction inhibitor oleamide, or via time-lapse microscopy of IEC-6 cells. Connexin 43 expression, trafficking, and localization were detected in cultured or primary enterocytes or mouse or human intestine by confocal microscopy and 35S-labeling, and gap junction communication was assessed using live microscopy with oleamide or connexin 43 siRNA. Results: Enterocytes expressed connexin 43 in vitro and in vivo, and exchanged fluorescent dye via gap junctions. Gap junction inhibition significantly reduced enterocyte migration in vitro and in vivo. NEC was associated with IFN release and loss of enterocyte connexin 43 expression. IFN inhibited enterocyte migration by reducing gap junction communication through the dephosphorylation and internalization of connexin 43. Gap junction inhibition significantly increased NEC severity, whereas reversal of the inhibitory effects of IFN on gap junction communication restored enterocyte migration after IFN exposure. Strikingly, IFN-/- mice were protected from the development of NEC, and showed restored connexin 43 expression and intestinal restitution. Conclusions: IFN inhibits enterocyte migration by preventing interenterocyte gap junction communication. Connexin 43 loss may provide insights into the development of NEC, in which restitution is impaired.
AB - Background & Aims: Necrotizing enterocolitis (NEC) is characterized by interferon-gamma (IFN-γ) release and inadequate intestinal restitution. Because enterocytes migrate together, mucosal healing may require interenterocyte communication via connexin 43-mediated gap junctions. We hypothesize that enterocyte migration requires interenterocyte communication, that IFN impairs migration by impairing connexin 43, and that impaired healing during NEC is associated with reduced gap junctions. Methods: NEC was induced in Swiss-Webster or IFN-/- mice, and restitution was determined in the presence of the gap junction inhibitor oleamide, or via time-lapse microscopy of IEC-6 cells. Connexin 43 expression, trafficking, and localization were detected in cultured or primary enterocytes or mouse or human intestine by confocal microscopy and 35S-labeling, and gap junction communication was assessed using live microscopy with oleamide or connexin 43 siRNA. Results: Enterocytes expressed connexin 43 in vitro and in vivo, and exchanged fluorescent dye via gap junctions. Gap junction inhibition significantly reduced enterocyte migration in vitro and in vivo. NEC was associated with IFN release and loss of enterocyte connexin 43 expression. IFN inhibited enterocyte migration by reducing gap junction communication through the dephosphorylation and internalization of connexin 43. Gap junction inhibition significantly increased NEC severity, whereas reversal of the inhibitory effects of IFN on gap junction communication restored enterocyte migration after IFN exposure. Strikingly, IFN-/- mice were protected from the development of NEC, and showed restored connexin 43 expression and intestinal restitution. Conclusions: IFN inhibits enterocyte migration by preventing interenterocyte gap junction communication. Connexin 43 loss may provide insights into the development of NEC, in which restitution is impaired.
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U2 - 10.1053/j.gastro.2007.03.029
DO - 10.1053/j.gastro.2007.03.029
M3 - Article
C2 - 17570214
AN - SCOPUS:34250003717
SN - 0016-5085
VL - 132
SP - 2395
EP - 2411
JO - Gastroenterology
JF - Gastroenterology
IS - 7
ER -