Interferon-γ Inhibits Intestinal Restitution by Preventing Gap Junction Communication Between Enterocytes

Cynthia L. Leaphart, Faisal Qureshi, Selma Cetin, Jun Li, Theresa Dubowski, Catherine Batey, Donna Beer-Stolz, Fengli Guo, Sandra A. Murray, David J. Hackam

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2395-2411
Number of pages17
JournalGastroenterology
Volume132
Issue number7
DOIs
StatePublished - Jun 1 2007

Fingerprint

Enterocytes
Gap Junctions
Connexin 43
Interferons
Necrotizing Enterocolitis
Communication
Interferon-gamma
Microscopy
Fluorescent Dyes
Confocal Microscopy
Small Interfering RNA
Intestines

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Interferon-γ Inhibits Intestinal Restitution by Preventing Gap Junction Communication Between Enterocytes. / Leaphart, Cynthia L.; Qureshi, Faisal; Cetin, Selma; Li, Jun; Dubowski, Theresa; Batey, Catherine; Beer-Stolz, Donna; Guo, Fengli; Murray, Sandra A.; Hackam, David J.

In: Gastroenterology, Vol. 132, No. 7, 01.06.2007, p. 2395-2411.

Research output: Contribution to journalArticle

Leaphart, CL, Qureshi, F, Cetin, S, Li, J, Dubowski, T, Batey, C, Beer-Stolz, D, Guo, F, Murray, SA & Hackam, DJ 2007, 'Interferon-γ Inhibits Intestinal Restitution by Preventing Gap Junction Communication Between Enterocytes', Gastroenterology, vol. 132, no. 7, pp. 2395-2411. https://doi.org/10.1053/j.gastro.2007.03.029
Leaphart, Cynthia L. ; Qureshi, Faisal ; Cetin, Selma ; Li, Jun ; Dubowski, Theresa ; Batey, Catherine ; Beer-Stolz, Donna ; Guo, Fengli ; Murray, Sandra A. ; Hackam, David J. / Interferon-γ Inhibits Intestinal Restitution by Preventing Gap Junction Communication Between Enterocytes. In: Gastroenterology. 2007 ; Vol. 132, No. 7. pp. 2395-2411.
@article{eb01200d90cb4dff93802d0205c1830c,
title = "Interferon-γ Inhibits Intestinal Restitution by Preventing Gap Junction Communication Between Enterocytes",
abstract = "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.",
author = "Leaphart, {Cynthia L.} and Faisal Qureshi and Selma Cetin and Jun Li and Theresa Dubowski and Catherine Batey and Donna Beer-Stolz and Fengli Guo and Murray, {Sandra A.} and Hackam, {David J.}",
year = "2007",
month = "6",
day = "1",
doi = "10.1053/j.gastro.2007.03.029",
language = "English (US)",
volume = "132",
pages = "2395--2411",
journal = "Gastroenterology",
issn = "0016-5085",
publisher = "W.B. Saunders Ltd",
number = "7",

}

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.

PY - 2007/6/1

Y1 - 2007/6/1

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.

UR - http://www.scopus.com/inward/record.url?scp=34250003717&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34250003717&partnerID=8YFLogxK

U2 - 10.1053/j.gastro.2007.03.029

DO - 10.1053/j.gastro.2007.03.029

M3 - Article

C2 - 17570214

AN - SCOPUS:34250003717

VL - 132

SP - 2395

EP - 2411

JO - Gastroenterology

JF - Gastroenterology

SN - 0016-5085

IS - 7

ER -