Spatio-temporal propagation of Ca2+ signals by cyclic ADP-ribose in 3T3 cells stimulated via purinergic P2Y receptors

被引:29
作者
Bruzzone, S
Kunerth, S
Zocchi, E
De Flora, A
Guse, AH
机构
[1] Univ Hosp Hamburg Eppendorf, Inst Biochem & Mol Biol 1, Ctr Med Expt, D-20246 Hamburg, Germany
[2] Univ Genoa, Dept Expt Med, Biochem Sect, I-16132 Genoa, Italy
[3] Univ Genoa, Ctr Excellence Biomed Res, I-16132 Genoa, Italy
基金
英国惠康基金;
关键词
cyclic ADP-ribose; Ca2+ signaling; 3T3; cell; CD38; signal transduction;
D O I
10.1083/jcb.200307016
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The role of cyclic ADP-ribose in the amplification of subcellular and global Ca2+ signaling upon stimulation of P2Y purinergic receptors was studied in 3T3 fibroblasts. Either (1) 3T3 fibroblasts (CD38(-) cells), (2) 3T3 fibroblasts preloaded by incubation with extracellular cyclic ADP-ribose (cADPR), (3) 3T3 fibroblasts microinjected with ryanodine, or (4) 3T3 fibroblasts transfected to express the ADP-ribosyl cyclase CD38 (CD38(+) cells) were used. Both preincubation with cADPR and CD38 expression resulted in comparable intracellular amounts of cyclic ADP-ribose (42.3 +/- 5.2 and 50.5 +/- 8.0 pmol/mg protein). P2Y receptor stimulation of CD38(-) cells yielded a small increase of intracellular Ca2+ concentration and a much higher Ca2+ signal in CD38-transfected cells, in cADPR-preloaded cells, or in cells microinjected with ryanodine. Confocal Ca2+ imaging revealed that stimulation of ryanodine receptors by cADPR or ryanodine amplified localized pacemaker Ca2+ signals with properties resembling Ca2+ quarks and triggered the propagation of such localized signals from the plasma membrane toward the internal environment, thereby initiating a global Ca2+ wave.
引用
收藏
页码:837 / 845
页数:9
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