Spatiotemporal laser inactivation of inositol 1,4,5-trisphosphate receptors using synthetic small-molecule probes
Spatiotemporal laser inactivation of inositol 1,4,5-trisphosphate receptors using synthetic small-molecule probes
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DOI：
10.1016/s1074-5521(03)00122-4
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发表时间：
2003-06-01
影响因子：
--
通讯作者：
Nagano, T
中科院分区：
文献类型：
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作者：
Inoue, T;Kikuchi, K;Nagano, T
孔雀石绿共轭肌醇1,4,5-三磷酸（MGIP(3)）在激光照射下诱导组织样品中IP3受体（IP3R）特异性失活。为了验证该方法在细胞Ca2+信号研究中的潜在用途，我们在单细胞水平上进行了激光失活，并表明IP3R在极高的时空分辨率下失活。在MGIP存在的情况下(3)，单个B淋巴瘤细胞IP3R的Ca2+释放功能随着激光照射时间的增加呈指数衰减，时间常数为3.4 s。此外，通过将激光照射限制在分化的PC12细胞的空间不同区域，IP3R的亚细胞失活被实现，正如本地呼叫信号的丢失所揭示的那样。这种仅在亚细胞区域内的IP3R的实时失活可能为研究Ca2+信号的时空动态提供了一种强有力的方法。
A malachite green-conjugated inositol 1,4,5-trisphosphate (MGIP(3)) induces specific inactivation Of IP3 receptor (IP3R) in tissue samples upon laser irradiation. To verify potential usefulness of the method for studies of cellular Ca2+ signaling, we conducted laser inactivation at the single-cell level and show that IP3R was inactivated with extremely high spatiotemporal resolution. In the presence of MGIP(3), the Ca2+ release function Of IP3R in single B lymphoma cells decayed exponentially with increasing duration of laser irradiation with a time constant of 3.4 s. Moreover, by confining laser irradiation to a spatially distinct region of differentiated PC12 cells, subcellular inactivation Of IP3R was attained, as revealed by a loss of local Call signal. Such real-time inactivation Of IP3R only within a subcellular region may provide a powerful method for investigating spatiotemporal dynamics of Ca2+ Signaling.