Cav3.1 (alpha1G) controls von Willebrand factor secretion in rat pulmonary microvascular endothelial cells.
The T-type Ca2+ channel Cav3.1 subunit is present in pulmonary microvascular endothelial cells (PMVECs), but not in pulmonary artery endothelial cells (PAECs). The present study sought to assess the role of Cav3.1 in thrombin-induced Weibel-Palade body exocytosis and consequent von Willebrand factor (VWF) release. In PMVECs and PAECs transduced with a green fluorescent protein (GFP)-tagged VWF chimera, we examined the real-time dynamics and secretory process of VWF-GFP-containing vesicles in response to thrombin and the cAMP-elevating agent isoproterenol. Whereas thrombin stimulated a progressive decrease in the number of VWF-GFP-containing vesicles in both cell types, isoproterenol only decreased the number of VWF-GFP-containing vesicles in PAECs. In PMVECs, thrombin-induced decrease in the number of VWF-GFP-containing vesicles was nearly abolished by the T-type Ca2+ channel blocker mibefradil as well as by Cav3.1 gene silencing with small hairpin RNA. Expression of recombinant Cav3.1 subunit in PAECs resulted in pronounced increase in thrombin-stimulated Ca2+ entry, which is sensitive to mibefradil. Together, these data indicate that VWF secretion from lung endothelial cells is regulated by two distinct pathways involving Ca2+ or cAMP, and support the hypothesis that activation of Cav3.1 T-type Ca2+ channels in PMVECs provides a unique cytosolic Ca2+ source important for Gq-linked agonist-induced VWF release.
Zhou
Chun
C
Center for Lung Biology, University of South Alabama College of Medicine, Mobile, AL 36688-0002, USA.
Chen
Hairu
H
Lu
Fengmin
F
Sellak
Hassan
H
Daigle
Jonathan A
JA
Alexeyev
Mikhail F
MF
Xi
Yaguang
Y
Ju
Jingfang
J
van Mourik
Jan A
JA
Wu
Songwei
S
D000818:Animals; D020747:Calcium Channels, T-Type / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D005089:Exocytosis / Q000187:drug effects; D008168:Lung / Q000098:blood supply* / Q000166:cytology; D020748:Mibefradil / Q000494:pharmacology; D008833:Microcirculation / Q000502:physiology; D034622:RNA Interference; D051381:Rats; D013917:Thrombin / Q000502:physiology; D020302:Weibel-Palade Bodies / Q000502:physiology; D014841:von Willebrand Factor / Q000378:metabolism*
D020747:Calcium Channels, T-Type; D014841:von Willebrand Factor; D020748:Mibefradil; D013917:Thrombin