Modification of intracellular Ca2+ dynamics by laser inactivation of inositol 1,4,5-trisphosphate receptor using membrane-permeant probes
Modification of intracellular Ca2+ dynamics by laser inactivation of inositol 1,4,5-trisphosphate receptor using membrane-permeant probes
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DOI：
10.1016/j.chembiol.2004.05.012
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发表时间：
2004-08-01
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：
Yogo, T;Kikuchi, K;Nagano, T
合成了一种具有膜渗透性的孔雀石绿偶联的IP 3类似物（MGIP（3）/PM）作为小分子卡利（smCALI）的探针，并在完整的鸡B细胞中检测了其对Ca 2+信号的影响。在用smCALI探针处理的DT 40 B细胞中，激光照射抑制IP 3诱导的响应于B细胞受体刺激的Ca 2+振荡，表明IP 3 R被急性灭活。然后，我们应用smCALI澄清的电容性Ca 2+内流（CCE）的机制，其中IP 3 R的参与已被建议。尽管smCALI使IP 3R失活，但毒胡萝卜素诱导的CCE仍不受影响，这提供了功能性IP 3R不是DT 40细胞中CCE所需的证据。这些结果证明了smCALI技术的潜力的IP 3R在复杂的细胞内Ca 2+动力学的作用的研究。
A membrane-permeant malachite green-conjugated IP3 analog (MGIP(3)/PM) was synthesized as a probe for small molecule-based CALI (smCALI), and its effect on the Ca2+ signaling in intact DT40 chicken B cells was examined. In DT40 B cells treated with the smCALI probe, laser irradiation inhibited IP3-induced Ca2+ oscillations in response to B cell receptor stimulation, demonstrating that IP3R was acutely inactivated. We then applied smCALI to clarify the mechanism of capacitative Ca2+ entry (CCE), in which involvement of IP3R has been suggested. Despite the inactivation of IP3R by smCALI, thapsigargin-induced CCE remained unaffected, providing evidence that functional IP3R is not required for CCE in DT40 cells. These results demonstrate the potency of the smCALI technique for the study of the roles of IP3R in complex intracellular Ca2+ dynamics.