Characterization of G protein-coupled receptors expressed by ECV304 human endothelial cells.
The aims of this study were to characterize G protein-coupled receptors endogenously expressed by ECV304 human endothelial cells, and to determine the utility of this transformed cell line as a vehicle for the expression of cloned receptors. Cellular responses to a broad range of agonists were determined by measuring changes in the intracellular content of second messengers (inositol phosphates and cyclic adenosine monophosphate). These studies identified H1 histamine receptors, P2U-purinoceptors and lysophosphatidic acid receptors which are functionally coupled to phosphoinositidase C. G protein-coupled receptors which bind adenosine (A2 receptor), calcitonin, and adrenaline (beta-adrenoceptor), and markedly stimulate adenylyl cyclase, are also endogenously expressed by ECV304. Agonists which did not stimulate ECV304 cells are: angiotensin II, angiotensin1-7, bombesin, bradykinin, desArg9-bradykinin, carbachol, endothelin-1, neurotensin, serotonin, substance K, substance P, thrombin and vasopressin. The rat Via vasopressin receptor was expressed by lipofection in two antibiotic-resistant clonal lines and expression confirmed by measuring agonist-induced changes in inostol phosphate production. We conclude that the ECV304 cell line is a suitable in vitro system to study the signal transduction pathways of some endogenous G protein-coupled receptors known to modulate endothelial function in vivo. ECV304 is also appropriate for the expression and functional characterization of cloned receptor proteins.
Howl
J
J
University of Wolverhampton, School of Health Sciences, UK. in5707@wlv.ac.uk
Mondszein
R M
RM
Wheatley
M
M
D000818:Animals; D002461:Cell Line, Transformed; D004730:Endothelium, Vascular / Q000502:physiology*; D019204:GTP-Binding Proteins / Q000502:physiology*; D018014:Gene Transfer Techniques; D006801:Humans; D051381:Rats; D011956:Receptors, Cell Surface / Q000819:agonists / Q000502:physiology*; D015290:Second Messenger Systems / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*
D011956:Receptors, Cell Surface; D019204:GTP-Binding Proteins