Extracellular ATP signaling and P2X nucleotide receptors in monolayers of primary human vascular endothelial cells

• DOI: 10.1152/ajpcell.01387.2000
• Publication Date: 2013-05-14T00:06:53Z
• Authors: Lisa M. Schwiebert; William C. Rice; Brian A. Kudlow; Amanda L. Taylor; Erik M. Schwiebert
• Keywords: Purinergic Signalling; P2Y receptor; Thapsigargin

ATP and its metabolites regulate vascular tone; however, the sources of released in beds are ill defined. As such, we tested hypothesis that all limbs an extracellular purinergic signaling system present endothelial cells: release, receptors, receptor-triggered signal transduction. Primary cultures human cells derived from multiple blood vessels were grown as monolayers studied using a bioluminescence detection assay for into medium. is constitutively exclusively across apical membrane under basal conditions. Hypotonic challenge or calcium agonists ionomycin thapsigargin stimulate release reversible regulated manner. To assess expression P2X receptor channel subtypes (P2XRs), performed degenerate RT-PCR, sequencing P2XR product, immunoblotting with subtype-specific antibodies. Results revealed 4 5 expressed abundantly by cell primary vessels. Together, these results suggest components autocrine loop exist microvasculature may allow “self-regulation” function modulation tone.