Astrocyte-endothelial cell calcium signals conveyed by two signalling pathways.
Astrocytes and endothelial cells are in close contact with each other at the blood-brain barrier, where important molecular transports take place. Despite these key morphological and functional properties, little is known regarding the dynamic signalling processes that occur between these two cell types. We investigated astrocyte-endothelial cell calcium signalling mechanisms in a coculture model prepared from primary rat cortical astrocytes and ECV304 cells. We used flash photolysis of caged inositol-trisphosphate (IP3) and gentle mechanical stimulation to trigger astrocyte-endothelial cell calcium signals and to investigate the underlying propagation mechanisms. Photolytically releasing IP3 in a single cell triggered increases in cytoplasmic calcium concentration that propagated between astrocytes and endothelial cells in either direction. These propagating calcium signals did not cross cell-free zones and were not affected by fast superfusion or by the purinergic inhibitors apyrase and suramin, indicating that they are communicated through an intracellular pathway in conjunction with gap junctions. Electrophysiological experiments confirmed a low degree of astrocyte-endothelial cell electrical cell-to-cell coupling. Mechanical stimulation of a single cell also triggered astrocyte-endothelial cell calcium signals but, in contrast to the former triggering mode, these signals crossed cell-free zones and were significantly inhibited by apyrase, thus indicating the involvement of an extracellular and purinergic messenger. Astrocyte-endothelial cell calcium signalling also occurred in cocultures prepared with astrocytes and primary rat brain capillary endothelial cells. We conclude that astrocytes and endothelial cells can exchange fast-acting calcium signals (time scale of seconds) that can be communicated through an intracellular/gap junctional pathway and an extracellular purinergic pathway.
Braet
K
K
Department of Physiology and Pathophysiology, Ghent University, De Pintelaan 185 (Blok B), B-9000 Ghent, Belgium.
Paemeleire
K
K
D'Herde
K
K
Sanderson
M J
MJ
Leybaert
L
L
D000818:Animals; D001253:Astrocytes / Q000502:physiology* / Q000648:ultrastructure; D001921:Brain / Q000166:cytology; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D002196:Capillaries / Q000166:cytology / Q000502:physiology; D002560:Cerebrovascular Circulation; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D005110:Extracellular Space / Q000378:metabolism; D017629:Gap Junctions / Q000502:physiology; D015544:Inositol 1,4,5-Trisphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D007425:Intracellular Membranes / Q000378:metabolism; D008854:Microscopy, Electron; D010782:Photolysis; D010812:Physical Stimulation; D011687:Purines / Q000378:metabolism; D051381:Rats; D015290:Second Messenger Systems / Q000502:physiology
D011687:Purines; C061868:inositol 1,4,5-trisphosphate 1-(2-nitrophenyl)ethyl ester; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium