Pertussis toxin induces angiogenesis in brain microvascular endothelial cells

J Neurosci Res. 2008 Sep;86(12):2624-40. doi: 10.1002/jnr.21716.

Abstract

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 approximately 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 >or=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 beta1 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.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Brain / blood supply*
  • Brain / drug effects*
  • Brain / physiology
  • Cells, Cultured
  • Cerebrovascular Circulation / drug effects
  • Cerebrovascular Circulation / genetics
  • Cerebrovascular Circulation / physiology
  • Endothelial Cells / drug effects*
  • Endothelial Cells / physiology
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / physiology
  • Gene Expression Regulation / drug effects
  • Inflammation Mediators / pharmacology
  • Mice
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Microcirculation / drug effects
  • Microcirculation / genetics
  • Microcirculation / physiology
  • Neovascularization, Physiologic / drug effects*
  • Neovascularization, Physiologic / genetics
  • Neovascularization, Physiologic / physiology
  • Oligonucleotide Array Sequence Analysis
  • Pertussis Toxin / pharmacology*

Substances

  • Inflammation Mediators
  • Pertussis Toxin