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Pertussis toxin induces angiogenesis in brain microvascular endothelial cells
Author(s) -
Lu Changming,
Pelech Steven,
Zhang Hong,
Bond Jeffrey,
Spach Karen,
Noubade Rajkumar,
Blankenhorn Elizabeth P.,
Teuscher Cory
Publication year - 2008
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.21716
Subject(s) - angiogenesis , pertussis toxin , phosphorylation , biology , signal transduction , phospholipase c , kinase , protein kinase a , microbiology and biotechnology , chemistry , g protein , cancer research
Pertussis toxin (PTX) is an ancillary adjuvant used to elicit experimental allergic encephalomyelitis (EAE), the principal autoimmune model of multiple sclerosis. One mechanism whereby PTX potentiates EAE is to increase blood–brain barrier (BBB) permeability. To elucidate further the mechanism of action of PTX on the BBB, we investigated the genomic and proteomic responses of isolated mouse brain endothelial cells (MBEC) following intoxication. Among ∼14,000 mouse genes tracked by cDNA microarray, 34 showed altered expression in response to PTX. More than one‐third of these genes have roles in angiogenesis. Accordingly, we show that intoxication of MBEC induces tube formation in vitro and angiogenesis in vivo. The global effect of PTX on signaling protein levels and phosphorylation in MBEC was investigated by using Kinex antibody microarrays. In total, 113 of 372 pan‐specific and 58 of 258 phospho‐site‐specific antibodies revealed changes ≥25% following intoxication. Increased STAT1 Tyr‐701 and Ser‐727 phosphorylation; reduced phosphorylation of the activating phospho‐sites in Erk1, Erk2, and MAPKAPK2; and decreased phosphorylation of arrestin β1 Ser‐412 and Hsp27 Ser‐82 were confirmed by Kinetworks multi‐immunoblotting. The importance of signal transduction pathways on PTX‐induced MBEC tube formation was evaluated pharmacologically. Inhibition of phospholipase C, MEK1, and p38 MAP kinase had little effect, whereas inhibition of cAMP‐dependent protein kinase, protein kinase C, and phosphatidylinositol 3‐kinase partially blocked tube formation. Taken together, these findings are consistent with the concept that PTX may lead to increased BBB permeability by altering endothelial plasticity and angiogenesis. © 2008 Wiley‐Liss, Inc.