Endotoxin increases intercellular resistance in microvascular endothelial cells by a tyrosine kinase pathway.
Gap junction communication between microvascular endothelial cells has been proposed to contribute to the coordination of microvascular function. Septic shock may attenuate microvascular cell-to-cell communication. We hypothesized that lipopolysaccharide (LPS) attenuates communication between microvascular endothelial cells derived from rat hindlimb skeletal muscle. Endothelial cells grown in monolayers expressed mRNA for connexin 37, 40, and 43. The expression of connexin 43 protein was confirmed, but connexin 40 protein was not detected by immunocytochemistry or immunoblot analysis. Intercellular resistance between cells of the monolayer, calculated using a Bessel function model, was increased from 3.3 to 5.3 MOmega by LPS. The effect was seen after 1 h of exposure and required a minimum concentration of 10 ng/ml. Intercellular resistance returned to normal 1 h following removal of LPS. Neither the response to LPS, nor its reversal, was blocked by the protein synthesis inhibitor cycloheximide (10 microg/ml). Pretreatment of monolayers with the tyrosine kinase inhibitors PP-2 (10 nM), lavendustin-C (1 microM), and geldanamycin (200 nM) prevented this LPS response; geldanamycin was also able to reverse the response. Inhibitors of MAP kinases, PD 98059 (5 microM) and SB 202190 (5 microM), and PKC (500 nM bisindolylmaleimide I) were unable to block the LPS response. We propose that LPS attenuates cell-to-cell communication through a signaling pathway that is tyrosine kinase dependent.
Lidington
D
D
Department of Medical Biophysics and University of Western Ontario, London, Canada.
Ouellette
Y
Y
Tyml
K
K
D000818:Animals; D002450:Cell Communication / Q000502:physiology*; D002478:Cells, Cultured; D017630:Connexins / Q000096:biosynthesis; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology*; D017629:Gap Junctions / Q000502:physiology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008833:Microcirculation; D011505:Protein-Tyrosine Kinases / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D015398:Signal Transduction
D017630:Connexins; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D011505:Protein-Tyrosine Kinases