EFFECT OF P2Y-AGONISTS ON ADENOSINE TRANSPORT IN CULTURED CHROMAFFIN CELLS

被引:40
作者
SEN, RP [1 ]
DELICADO, EG [1 ]
CASTRO, E [1 ]
MIRASPORTUGAL, MT [1 ]
机构
[1] UNIV COMPLUTENSE MADRID,FAC VET,DEPT BIOQUIM,E-28040 MADRID,SPAIN
关键词
ADENOSINE TRANSPORT; CHROMAFFIN CELLS; DIADENOSINE TETRAPHOSPHATE; NUCLEOSIDE TRANSPORT; P2Y-AGONISTS; PURINERGIC RECEPTORS;
D O I
10.1111/j.1471-4159.1993.tb03192.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Adenosine transport in cultured chromaffin cells was inhibited by purinergic P2y-receptor agonists without significant changes in the affinity constant, the values being between 1 +/- 0.4 and 1.6 +/- 0.6 muM. The V(max) parameter was modified significantly, being 40 +/- 1.0, 26 +/- 5.0, 32 +/- 3.0, and 22 +/- 4.7 pmol/10(6) cells/min for control, adenosine-5'O-(2-thiodiphosphate), 5'-adenylylimidodiphosphate, and p1,p4 -di(adenosine-5'-) tetraphosphate (Ap4A) (100 muM for every effector), respectively. AP4A, a physiological ligand for P2y receptors in chromaffin cells, showed the highest inhibitory effect (45%). This transport inhibition is explained by an increase in the cytosolic Ca2+ concentration ([Ca2+]i) and the activation of protein kinase C (PKC). Experiments of [Ca2+]i measurement with the fura-2 technique showed that P2y agonists, as well as bradykinin, were able to increase [Ca2+]i, this effect being independent of the presence of extracellular Ca2+. The peptide bradykinin, determined to be coupled to phosphatidylinositol hydrolysis and internal Ca2+ mobilization in chromaffin cells, exhibited a behavior similar to that of P2y agonists in adenosine transport inhibition (39%). P2y agonists and bradykinin increased PKC activity associated with the membrane fraction (about 50% increase in particulate PKC activity with respect to controls). The present studies suggest that adenosine transport is regulated by P2y-purinergic receptors mediated via Ca2+ mobilization and PKC activation.
引用
收藏
页码:613 / 619
页数:7
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