Lipopolisakkarid enterositlerdeki ara baǧlanti proteini konneksin-43′ü etkileyerek hücreler arasi iletişimi bozmaktadir

Translated title of the contribution: Lipopolysaccharide impairs cell-to-cell communication by affecting the gap junction protein connexin-43 in enterocytes

Orkan Ergün, Faisal G. Qureshi, Catherine Baty, Jun Li, Henri Ford, David J. Hackam

Research output: Contribution to journalArticle

Abstract

Aim: Communication between enterocytes is likely to be essential for the maintenance of gut barrier integrity. In many cells such as neurons, intercellular communication occurs through the gap junction (GJ) protein connexin-43 (Cx43), and phosphorylation ofCx43 (pCx43) disrupts GJ function. However, the in vivo effects of LPS on GJ expression and phosphorylation in enterocytes are undefined. We hypothesized that LPS modulates connexin-43 expression and phosphorylation in enterocytes in-vitro. Methods: Inter-enterocyte communication was measured by microinjecting intestinal epithelial cells (IEC-6) with lucifer-yellow (LY) and a larger molecule "rhodamine-dextrane (Rd-D). The specificity of GJ function was assessed by pretreatment of cells with "oleamide", a specific inhibitor of GJ function. Images were obtained by laser confocal microscopy. The expression and distribution of Cx-43 and its phosphorylation (pCx-43) were assessed in IEC-6 cells following treatment with LPS (50 μg/ml 24h) by Western blot and confocal microscopy. Results: Enterocyte injections allowed passage of LY but not Rd-dextran to adjoining cells, suggesting GJ function. Oleamide treatment inhibited the passage of LY to the adjacent cells. LPS significantly increased enterocyte pCx-43 (density rel to actin ctrl: 0.3±0.1 vs LPS 0.9±0.2, p<0.05) but not total Cx-43. LPS significantly increased accumulation of pCX-43 between cells. Conclusion: LPS impairs the function of GJ by increasing the phosphorilation of Cx-43. This may provide insights into maintanence of barrier function by interenterocyte function.

Original languageTurkish
Pages (from-to)53-58
Number of pages6
JournalCocuk Cerrahisi Dergisi
Volume19
Issue number2
StatePublished - Dec 1 2005
Externally publishedYes

Fingerprint

Connexin 43
Connexins
Enterocytes
Gap Junctions
Cell Communication
Lipopolysaccharides
Phosphorylation
Confocal Microscopy
Rhodamines
Dextrans
Actins
Western Blotting
Epithelial Cells
Maintenance
Neurons
Injections
Therapeutics
lucifer yellow

Keywords

  • Connexin-43
  • Gap junctions
  • Mucosal barrier
  • Necrotizing enterocolitis

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health
  • Surgery

Cite this

Lipopolisakkarid enterositlerdeki ara baǧlanti proteini konneksin-43′ü etkileyerek hücreler arasi iletişimi bozmaktadir. / Ergün, Orkan; Qureshi, Faisal G.; Baty, Catherine; Li, Jun; Ford, Henri; Hackam, David J.

In: Cocuk Cerrahisi Dergisi, Vol. 19, No. 2, 01.12.2005, p. 53-58.

Research output: Contribution to journalArticle

Ergün, Orkan ; Qureshi, Faisal G. ; Baty, Catherine ; Li, Jun ; Ford, Henri ; Hackam, David J. / Lipopolisakkarid enterositlerdeki ara baǧlanti proteini konneksin-43′ü etkileyerek hücreler arasi iletişimi bozmaktadir. In: Cocuk Cerrahisi Dergisi. 2005 ; Vol. 19, No. 2. pp. 53-58.
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T1 - Lipopolisakkarid enterositlerdeki ara baǧlanti proteini konneksin-43′ü etkileyerek hücreler arasi iletişimi bozmaktadir

AU - Ergün, Orkan

AU - Qureshi, Faisal G.

AU - Baty, Catherine

AU - Li, Jun

AU - Ford, Henri

AU - Hackam, David J.

PY - 2005/12/1

Y1 - 2005/12/1

N2 - Aim: Communication between enterocytes is likely to be essential for the maintenance of gut barrier integrity. In many cells such as neurons, intercellular communication occurs through the gap junction (GJ) protein connexin-43 (Cx43), and phosphorylation ofCx43 (pCx43) disrupts GJ function. However, the in vivo effects of LPS on GJ expression and phosphorylation in enterocytes are undefined. We hypothesized that LPS modulates connexin-43 expression and phosphorylation in enterocytes in-vitro. Methods: Inter-enterocyte communication was measured by microinjecting intestinal epithelial cells (IEC-6) with lucifer-yellow (LY) and a larger molecule "rhodamine-dextrane (Rd-D). The specificity of GJ function was assessed by pretreatment of cells with "oleamide", a specific inhibitor of GJ function. Images were obtained by laser confocal microscopy. The expression and distribution of Cx-43 and its phosphorylation (pCx-43) were assessed in IEC-6 cells following treatment with LPS (50 μg/ml 24h) by Western blot and confocal microscopy. Results: Enterocyte injections allowed passage of LY but not Rd-dextran to adjoining cells, suggesting GJ function. Oleamide treatment inhibited the passage of LY to the adjacent cells. LPS significantly increased enterocyte pCx-43 (density rel to actin ctrl: 0.3±0.1 vs LPS 0.9±0.2, p<0.05) but not total Cx-43. LPS significantly increased accumulation of pCX-43 between cells. Conclusion: LPS impairs the function of GJ by increasing the phosphorilation of Cx-43. This may provide insights into maintanence of barrier function by interenterocyte function.

AB - Aim: Communication between enterocytes is likely to be essential for the maintenance of gut barrier integrity. In many cells such as neurons, intercellular communication occurs through the gap junction (GJ) protein connexin-43 (Cx43), and phosphorylation ofCx43 (pCx43) disrupts GJ function. However, the in vivo effects of LPS on GJ expression and phosphorylation in enterocytes are undefined. We hypothesized that LPS modulates connexin-43 expression and phosphorylation in enterocytes in-vitro. Methods: Inter-enterocyte communication was measured by microinjecting intestinal epithelial cells (IEC-6) with lucifer-yellow (LY) and a larger molecule "rhodamine-dextrane (Rd-D). The specificity of GJ function was assessed by pretreatment of cells with "oleamide", a specific inhibitor of GJ function. Images were obtained by laser confocal microscopy. The expression and distribution of Cx-43 and its phosphorylation (pCx-43) were assessed in IEC-6 cells following treatment with LPS (50 μg/ml 24h) by Western blot and confocal microscopy. Results: Enterocyte injections allowed passage of LY but not Rd-dextran to adjoining cells, suggesting GJ function. Oleamide treatment inhibited the passage of LY to the adjacent cells. LPS significantly increased enterocyte pCx-43 (density rel to actin ctrl: 0.3±0.1 vs LPS 0.9±0.2, p<0.05) but not total Cx-43. LPS significantly increased accumulation of pCX-43 between cells. Conclusion: LPS impairs the function of GJ by increasing the phosphorilation of Cx-43. This may provide insights into maintanence of barrier function by interenterocyte function.

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KW - Necrotizing enterocolitis

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