title: Critical role of TRPP2 and TRPC1 channels in stretch-induced injury of blood-brain barrier endothelial cells.
abstract: The microvessels of the brain are very sensitive to mechanical stresses such as observed in traumatic brain injury (TBI). Such stresses can quickly lead to dysfunction of the microvessel endothelial cells, including disruption of blood-brain barrier (BBB). It is now evident that elevation of cytosolic calcium levels ([Ca2+]i) can compromise the BBB integrity, however the mechanism by which mechanical injury can produce a [Ca2+]i increase in brain endothelial cells is unclear. To assess the effects of mechanical/stretch injury on [Ca2+]i signaling, mouse brain microvessel endothelial cells (bEnd3) were grown to confluency on elasticized membranes and [Ca2+]i monitored using fura 2 fluorescence imaging. Application of an injury, using a pressure/stretch pulse of 50 ms, induced a rapid transient increase in [Ca2+]i. In the absence of extracellular Ca2+, the injury-induced [Ca2+]i transient was greatly reduced, but not fully eliminated, while unloading of Ca2+ stores by thapsigargin treatment in the absence of extracellular Ca2+ abolished the injury transient. Application of LOE-908 and amiloride, TRPC and TRPP2 channel blockers, respectively, both reduced the transient [Ca2+]i increase. Further, siRNA knockdown assays directed at TRPC1 and TRPP2 expression also resulted in a reduction of the injury-induced [Ca2+]i response. In addition, stretch injury induced increases of NO production and actin stress fiber formation, both of which were markedly reduced upon treatment with LOE908 and/or amiloride. We conclude that mechanical injury of brain endothelial cells induces a rapid influx of calcium, mediated by TRPC1 and TRPP2 channels, which leads to NO synthesis and actin cytoskeletal rearrangement.
authors.affiliation: Department of Integrative Biology & Pharmacology, The University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Berrout
authors.forename: Min
authors.initials: M
authors.lastname: Jin
authors.forename: Roger G
authors.initials: RG
authors.lastname: O'Neil
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D001812:Blood-Brain Barrier / Q000166:cytology* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D013314:Stress, Mechanical; D050052:TRPC Cation Channels / Q000378:metabolism*; D050396:TRPP Cation Channels / Q000378:metabolism*
chemical_list: D002118:Calcium; D009569:Nitric Oxide; D050052:TRPC Cation Channels; D050396:TRPP Cation Channels; C096351:transient receptor potential cation channel, subfamily C, member 1