Differential involvement of thrombin receptors in Ca2+ release from two different intracellular stores in human platelets

被引:41
作者
Jardin, Isaac
Ben Amor, Nidhal
Bartegi, Ahgleb
Pariente, Jose A.
Salido, Gines M.
Rosado, Juan A. [1 ]
机构
[1] Univ Extremadura, Dept Physiol, Cell Physiol Res Grp, Caceres 10071, Spain
[2] Inst Super Biotechnol, Unite Rech Biochim, Monastir, Tunisia
关键词
acidic organelles; calcium; dense tubular system (DST); platelet; protease-activated receptor 1 (PAR-1); thrombin;
D O I
10.1042/BJ20060888
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
Physiological agonists increase cytosolic free Ca2+ concentration to regulate a number of cellular processes. The platelet thrombin receptors, PAR (protease-activated receptor) 1 PAR-4 and GPIb-IX-V (glycoprotein Ib-IX-V) have been described as potential contributors of thrombin-induced platelet aggregation. Platelets present two separate Ca2+ stores, the DTS (dense tubular system) and acidic organelles, differentiated by the distinct sensitivity of their respective SERCAs (sarcoplasmic/endoplasmic-reticulum Ca2+-ATPases) to TG (thapsigargin) and TBHQ [2,5-di-(tert-butyl) -1,4-hydroquinone]. However, the involvement of the thrombin receptors in Ca2+ release front each Ca2+ store remains unknown. Depletion of the DTS using ADP, which releases Ca2+ solely from the DTS, in combination with 10 nM TG, to selectively inhibit SERCA2 located on the DTS reduced Ca2+ release evoked by the PAR-1 agonist, SFLLRN, and the PAR-4 agonist, AYPGKF, by 80 and 50% respectively. Desensitization of PAR-1 and PAR-4 or pre-treatment with the PAR-1 and PAR-4 antagonists SCH 79797 and tcY-NH2 reduced Ca2+ mobilization induced by thrombin, and depletion of the DTS after desensitization or blockade of PAR-1 and PAR-4 had no significant effect on Ca2+ release stimulated by thrombin through the GPIb-IX-V receptor. Converse experiments showed that depletion of the acidic stores using TBHQ reduced Ca2+ release evoked by SFLLRN or AYPGKF, by 20 and 50% respectively, and abolished thrombin-stimulated Ca2+ release through the GPIb-IX-V receptor when PAR-1 and PAR-4 had been desensitized on blocked. Our results indicate that thrombin-induced activation of PAR-1 and PAR-4 evokes Ca2+ release front both Ca2+ stores, while activation of GPIb-IX-V by thrombin releases Ca2+ solely from the acidic compartments in human platelets.
引用
收藏
页码:167 / 174
页数:8
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