Moccia F, Nusco G A, Lim D, Ercolano E, Gragnaniello G, Brown E R, Santella L
Laboratory of Cell Biology, Stazione Zoologica Anton Dohrn, Villa Comunale, 80121 Naples, Italy.
Pflugers Arch. 2003 Aug;446(5):541-52. doi: 10.1007/s00424-003-1076-1. Epub 2003 May 16.
Cyclic ADP-ribose (cADPr) is a second messenger that regulates intracellular free [Ca2+] (Ca2+) in a variety of cell types, including immature oocytes from the starfish Astropecten auranciacus. In this study, we employed confocal laser scanning microscopy and voltage clamp techniques to investigate the source of the cADPr-elicited Ca2+ wave originating from the cortical Ca2+ patches we have described previously. The Ca2+ swing was accompanied by a membrane current with a reversal potential of approximately +20 mV. Decreasing external Na+ almost abolished the current without affecting the Ca2+ response. Removal of extracellular Ca2+ altered neither the Ca2+ transient nor the ionic current, nor did the holding potential exert any effect on the Ca2+ wave. Both the Ca2+ response and the membrane current were abolished when BAPTA, ruthenium red or 8-NH(2)-cADPr were preinjected into the oocytes, while perfusion with ADPr did not elicit any Ca2+ increase or ionic current. However, elevating Ca2+ by uncaging Ca2+ from nitrophenyl- (NP-EGTA) or by photoliberating inositol 1,4,5-trisphosphate (InsP(3)) induced an ionic current with biophysical properties similar to that elicited by cADPr. These results suggest that cADPr activates a Ca2+ wave by releasing Ca2+ from intracellular ryanodine receptors and that the rise in Ca2+ triggers a non-selective monovalent cation current that does not seem to contribute to the global Ca2+ elevation.
环磷酸腺苷核糖(cADPr)是一种第二信使,可调节多种细胞类型中的细胞内游离[Ca2+]([Ca2+]i),包括来自海星橙黄疣海星的未成熟卵母细胞。在本研究中,我们采用共聚焦激光扫描显微镜和电压钳技术,研究了先前描述的源自皮质Ca2+斑的cADPr引发的Ca2+波的来源。Ca2+波动伴随着膜电流,其反转电位约为+20 mV。降低细胞外Na+几乎消除了电流,而不影响Ca2+反应。去除细胞外Ca2+既不改变Ca2+瞬变也不改变离子电流,保持电位对Ca2+波也没有任何影响。当将BAPTA、钌红或8-NH(2)-cADPr预先注射到卵母细胞中时,Ca2+反应和膜电流均被消除,而用ADPr灌注未引起任何[Ca2+]i增加或离子电流。然而,通过从硝基苯基-(NP-EGTA)中释放Ca2+或通过光释放肌醇1,4,5-三磷酸(InsP(3))来升高[Ca2+]i会诱导出一种离子电流,其生物物理特性与cADPr引发的离子电流相似。这些结果表明,cADPr通过从细胞内兰尼碱受体释放Ca2+来激活Ca2+波,并且[Ca2+]i的升高触发了一种非选择性单价阳离子电流,该电流似乎对整体Ca2+升高没有贡献。