Characterization of the P2 receptors on the human umbilical vein endothelial cell line ECV304

被引:25
作者
Conant, AR
Fisher, MJ
McLennan, AG
Simpson, AWM [1 ]
机构
[1] Univ Liverpool, Dept Human Anat & Cell Biol, Liverpool L69 3GE, Merseyside, England
[2] Univ Liverpool, Sch Biol Sci, Liverpool L69 3GE, Merseyside, England
基金
英国惠康基金;
关键词
adenine dinucleotides; diadenosine polyphosphate; ATP; UTP; Ca2+; human; endothelial cells; P2; receptor;
D O I
10.1038/sj.bjp.0702082
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
1 To characterize the P2 receptors present on the human umbilical vein endothelial-derived cell line, ECV304, cytosolic Ca2+, ([Ca2+](c)), responses were recorded in single cells and in cell suspensions to a series of nucleotides and nucleotide agonists. 2 Concentration response curves were obtained in fura-2-loaded ECV304 cell suspensions, with EC50 values of 4.2 mu M for ATP, 2.5 mu M for UTP and 14 mu M for adenosine-5'-O-(3-thio)triphosphate (ATP gamma S). EC50 values for 2-methylthioATP, ADP, adenosine-5'-O-(2-thio)diphosphate (ADP beta S) and AMP were 0.5 mu M, 3.5 mu M, 15 mu M and 4.7 mu M respectively, but maximal [Ca2+](c) responses were less than those produced by a maximal addition of ATP/UTP. ECV304 cells were unresponsive to UDP and beta,gamma,methyleneATP. 3 Cross-desensitization studies on ECV304 cells suggested that ATP and UTP recognized the same receptor. However, ADP recognized a receptor distinct from the UTP-sensitive receptor and AMP recognized a third distinct receptor. 4 ECV304 [Ca2+](c) responses to 2-methylthioATP were inhibited in the presence of 30 mu M pyridoxalphosphate-6-azophenyl-2',4'-dis acid (PPADS), whereas [Ca2+](c) responses to UTP were unaffected by this treatment. 5 ECV304 cells responded to the diadenosine polyphosphate Ap,A with rises in [Ca2+](c). Apparent responses to Ap(4)A, Ap(5)A and Ap(6)A, were shown to be due to a minor nucleotide contaminant that could be removed by pre-treatment of the diadenosine samples with either alkaline phosphatase or apyrase. 6 ECV304 cells display a pharmacology consistent with the presence of at least two P2 receptors; a P2Y(2) receptor insensitive to the diadenosine polyphosphates and a P2Y(1) receptor sensitive to Ap(3)A. In addition, ECV304 cells respond to AMP with increases in [Ca2+](c) via an as yet uncharacterized receptor.
引用
收藏
页码:357 / 364
页数:8
相关论文
共 41 条
[1]   The regulation of vascular function by P2 receptors: multiple sites and multiple receptors [J].
Boarder, MR ;
Hourani, SMO .
TRENDS IN PHARMACOLOGICAL SCIENCES, 1998, 19 (03) :99-107
[2]   PPADS - AN ANTAGONIST AT ENDOTHELIAL P-2Y-PURINOCEPTORS BUT NOT P-2U-PURINOCEPTORS [J].
BROWN, C ;
TANNA, B ;
BOARDER, MR .
BRITISH JOURNAL OF PHARMACOLOGY, 1995, 116 (05) :2413-2416
[3]  
Burnstock Geoffrey, 1996, V198, P1
[4]   VASOMOTOR ACTIVITY OF DIADENOSINE TRIPHOSPHATE AND DIADENOSINE TETRAPHOSPHATE IN ISOLATED ARTERIES [J].
BUSSE, R ;
OGILVIE, A ;
POHL, U .
AMERICAN JOURNAL OF PHYSIOLOGY, 1988, 254 (05) :H828-H832
[5]   SINGLE-CELL FURA-2 MICROFLUOROMETRY REVEALS DIFFERENT PURINOCEPTOR SUBTYPES COUPLED TO CA2+ INFLUX AND INTRACELLULAR CA2+ RELEASE IN BOVINE ADRENAL CHROMAFFIN AND ENDOTHELIAL-CELLS [J].
CASTRO, E ;
TOME, AR ;
MIRASPORTUGAL, MT ;
ROSARIO, LM .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1994, 426 (06) :524-533
[6]   MOLECULAR-CLONING AND FUNCTIONAL-ANALYSIS OF A NOVEL P-2 NUCLEOTIDE RECEPTOR [J].
CHANG, KG ;
HANAOKA, K ;
KUMADA, M ;
TAKUWA, Y .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (44) :26152-26158
[7]   PPADS and suramin as antagonists at cloned P-2Y- and P-2U-purinoceptors [J].
Charlton, SJ ;
Brown, CA ;
Weisman, GA ;
Turner, JT ;
Erb, L ;
Boarder, MR .
BRITISH JOURNAL OF PHARMACOLOGY, 1996, 118 (03) :704-710
[8]   Cloned and transfected P2Y(4) receptors: Characterization of a suramin and PPADS-insensitive response to UTP [J].
Charlton, SJ ;
Brown, CA ;
Weisman, GA ;
Turner, JT ;
Erb, L ;
Boarder, MR .
BRITISH JOURNAL OF PHARMACOLOGY, 1996, 119 (07) :1301-1303
[9]   Characterization of signaling pathways of P-2Y and P-2U purinoceptors in bovine pulmonary artery endothelial cells [J].
Chen, BC ;
Lee, CM ;
Lee, YT ;
Lin, WW .
JOURNAL OF CARDIOVASCULAR PHARMACOLOGY, 1996, 28 (02) :192-199
[10]   Cloning, functional expression and tissue distribution of the human P2Y(6) receptor [J].
Communi, D ;
Parmentier, M ;
Boeynaems, JM .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1996, 222 (02) :303-308