title: Permeabilization in a cerebral endothelial barrier model by pertussis toxin involves the PKC effector pathway and is abolished by elevated levels of cAMP.
abstract: Respiratory tract infections caused by Bordetella pertussis are occasionally accompanied by severe neurologic disorders and encephalopathies. For these sequelae to occur the integrity of cerebral barriers needs to be compromised. The influence of pertussis toxin, a decisive virulence factor in the pathogenesis of pertussis disease, on barrier integrity was investigated in model systems for blood-liquor (epithelial) and blood-brain (endothelial) barriers. While pertussis toxin did not influence the barrier function in Plexus chorioideus model systems, the integrity of cerebral endothelial monolayers was severely compromised. Cellular intoxication by pertussis toxin proceeds via ADP-ribosylation of alpha-G(i) proteins, which not only interferes with the homeostatic inhibitory regulation of adenylate cyclase stimulation but also results in a modulation of the membrane receptor coupling. Increasing intra-endothelial cAMP levels by employing cholera toxin or forskolin even inhibited the pertussis toxin-induced permeabilization of endothelial barriers. Therefore, pertussis-toxin-induced permeabilization has to be mediated via a cAMP-independent pathway. To investigate potential signalling pathways we employed several well established cellular drugs activating or inhibiting central effectors of signal transduction pathways, such as phosphatidylinositol 3-kinase, adenylate cyclase, phospholipase C, myosin light chain kinase and protein kinase C. Only inhibitors and activators of protein kinase C and phosphatidylinositol 3-kinase affected the pertussis toxin-induced permeability. In summary, we conclude that permeabilization of cerebral endothelial monolayers by pertussis toxin does not depend on elevated cAMP levels and proceeds via the phosphokinase C pathway.
authors.affiliation: Institut für Infektiologie - Zentrum für Molekularbiologie der Entzündung (ZMBE), Universitätsklinikum Münster, Von-Esmarch-Str. 56, 48149 Münster, Germany.
authors.forename: Kerstin E
authors.initials: KE
authors.lastname: Brückener
authors.forename: Ali
authors.initials: A
authors.lastname: el Bayâ
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Galla
authors.forename: M Alexander
authors.initials: MA
authors.lastname: Schmidt
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects* / Q000502:physiology*; D002199:Capillary Permeability / Q000187:drug effects; D002478:Cells, Cultured; D002831:Choroid Plexus / Q000187:drug effects / Q000502:physiology; D046508:Culture Techniques; D000242:Cyclic AMP / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000503:physiopathology; D008959:Models, Neurological; D009422:Nervous System Diseases / Q000209:etiology; D037342:Pertussis Toxin / Q000633:toxicity*; D011493:Protein Kinase C / Q000378:metabolism*; D012756:Sheep; D015398:Signal Transduction / Q000187:drug effects; D034421:Sus scrofa; D014917:Whooping Cough / Q000150:complications
chemical_list: D000242:Cyclic AMP; D037342:Pertussis Toxin; D011493:Protein Kinase C