Alpha1G T-type calcium channel selectively regulates P-selectin surface expression in pulmonary capillary endothelium.
Regulated P-selectin surface expression provides a rapid measure for endothelial transition to a proinflammatory phenotype. In general, P-selectin surface expression results from Weibel-Palade body (WPb) exocytosis. Yet, it is unclear whether pulmonary capillary endothelium possesses WPbs or regulated P-selectin surface expression and, if so, how inflammatory stimuli initiate exocytosis. We used immunohistochemistry, immunofluorescence labeling, ultrastructural assessment, and an isolated perfused lung model to demonstrate that capillary endothelium lacks WPbs but possesses P-selectin. Thrombin stimulated P-selectin surface expression in both extra-alveolar vessel and alveolar capillary endothelium. Only in capillaries was the thrombin-stimulated P-selectin surface expression considerably mitigated by pharmacologic blockade of the T-type channel or genetic knockout of the T-type channel alpha(1G)-subunit. Depolarization of endothelial plasma membrane via high K(+) perfusion capable of eliciting cytosolic Ca(2+) transients also provoked P-selectin surface expression in alveolar capillaries that was abolished by T-type channel blockade or alpha(1G) knockout. Our findings reveal an intracellular WPb-independent P-selectin pool in pulmonary capillary endothelium, where the regulated P-selectin surface expression is triggered by Ca(2+) transients evoked through activation of the alpha(1G) T-type channel.
Zhou
Chun
C
Center for Lung Biology, University of South Alabama College of Medicine, Mobile, Alabama 36688-0002, USA.
Chen
Hairu
H
King
Judy A
JA
Sellak
Hassan
H
Kuebler
Wolfgang M
WM
Yin
Jun
J
Townsley
Mary I
MI
Shin
Hee-Sup
HS
Wu
Songwei
S
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000378:metabolism; D020747:Calcium Channels, T-Type / Q000235:genetics / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology; D042783:Endothelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000648:ultrastructure; D005089:Exocytosis / Q000502:physiology; D006801:Humans; D008168:Lung / Q000098:blood supply* / Q000648:ultrastructure; D008297:Male; D020748:Mibefradil / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020302:Weibel-Palade Bodies / Q000378:metabolism / Q000648:ultrastructure; D014841:von Willebrand Factor / Q000235:genetics / Q000378:metabolism
C121855:CACNA1G protein, human; C498197:Cacna1g protein, mouse; D002121:Calcium Channel Blockers; D020747:Calcium Channels, T-Type; D019007:P-Selectin; D021122:Protein Subunits; D014841:von Willebrand Factor; D020748:Mibefradil; D002118:Calcium