Low extracellular Ca2+ conditions induce an increase in brain endothelial permeability that involves intercellular Ca2+ waves.
The intracellular calcium concentration ([Ca(2+)](i)) is an important factor determining the permeability of endothelial barriers including the blood-brain barrier (BBB). However, nothing is known concerning the effect of spatially propagated intercellular Ca(2+) waves (ICWs). The propagation of ICWs relies in large part on channels formed by connexins that are present in endothelia. We hypothesized that ICWs may result in a strong disturbance of endothelial function, because the [Ca(2+)](i) changes are coordinated and involve multiple cells. Thus, we aimed to investigate the effect of ICWs on endothelial permeability. ICW activity was triggered in immortalized and primary brain endothelial cells by lowering the extracellular Ca(2+) concentration. Low extracellular Ca(2+) increased the endothelial permeability and this was significantly suppressed by buffering [Ca(2+)](i) with BAPTA-AM, indicating a central role of [Ca(2+)](i) changes. The endothelial permeability increase was furthermore inhibited by the connexin channel blocking peptide Gap27, which also blocked the ICWs, and by inhibiting protein kinase C (PKC), Ca(2+)/calmodulin-dependent kinase II (CaMKII) and actomyosin contraction. We compared these observations with the [Ca(2+)](i) changes and permeability alterations provoked by the inflammatory agent bradykinin (BK), which triggers oscillatory [Ca(2+)](i) changes without wave activity. BK-associated [Ca(2+)](i) changes and the endothelial permeability increase were significantly smaller than those associated with ICWs, and the permeability increase was not influenced by inhibition of PKC, CaMKII or actomyosin contraction. We conclude that ICWs significantly increase endothelial permeability and therefore, the connexins that underlie wave propagation form an interesting target to limit BBB alterations. This article is part of a Special Issue entitled Electrical Synapses.
Department of Basic Medical Sciences, Physiology Group, Ghent University, De Pintelaan 185, Building B, 9000 Ghent, Belgium.
Marijke
M
De Bock
Maxime
M
Culot
Nan
N
Wang
Anaelle
A
da Costa
Elke
E
Decrock
Mélissa
M
Bol
Geert
G
Bultynck
Romeo
R
Cecchelli
Luc
L
Leybaert
D000205:Actomyosin / Q000502:physiology; D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D015153:Blotting, Western; D001920:Bradykinin / Q000494:pharmacology; D001921:Brain / Q000502:physiology*; D002118:Calcium / Q000502:physiology*; D020013:Calcium Signaling / Q000502:physiology*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000502:physiology; D002478:Cells, Cultured; D017630:Connexins / Q000494:pharmacology; D003599:Cytoskeleton / Q000502:physiology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000502:physiology*; D005110:Extracellular Space / Q000187:drug effects / Q000502:physiology; D017629:Gap Junctions / Q000187:drug effects; D059906:Neuroimaging; D009842:Oligopeptides; D010539:Permeability; D011493:Protein Kinase C / Q000502:physiology; D051381:Rats
D017630:Connexins; D009842:Oligopeptides; C455880:gap 27 peptide; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D000205:Actomyosin; D011493:Protein Kinase C; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D001920:Bradykinin; D002118:Calcium