Apoptosis induction-related cytosolic calcium responses revealed by the dual FRET imaging of calcium signals and caspase-3 activation in a single cell
Apoptosis induction-related cytosolic calcium responses revealed by the dual FRET imaging of calcium signals and caspase-3 activation in a single cell
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单细胞中钙信号和 caspase-3 激活的双重 FRET 成像揭示了细胞凋亡诱导相关的胞质钙反应
DOI：
10.1016/j.bbrc.2015.02.045
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Mikoshiba K.
中科院分区：
文献类型：
--
作者：
Miyamoto A;Miyauchi H;Kogure T;Miyawaki A;Michikawa T;Mikoshiba K.
刺激诱导的细胞内Ca 2+浓度的变化控制细胞命运的决定，包括凋亡。然而，与凋亡诱导相关的胞质Ca 2+信号的精确模式仍然未知。我们已经开发了一种新的基因编码的传感器激活的半胱天冬酶-3，可以与基因编码的传感器的Ca 2+浓度的组合应用，并建立了一个双重成像系统，使成像的细胞溶质Ca 2+信号和半胱天冬酶-3激活，这是一个指标，在同一个细胞中。使用这个系统，我们确定了细胞内钙信号的差异凋亡和存活的DT 40 B淋巴细胞后B细胞受体（BCR）的刺激。在存活的细胞中，BCR刺激诱发更大的初始Ca 2+尖峰随后由一个更大的持续升高的Ca 2+浓度比那些在凋亡细胞; BCR刺激也导致重复的瞬时Ca 2+尖峰，这是由Ca 2+从细胞外空间的流入介导的。我们的研究结果表明，在同一个细胞中观察Ca 2+信号和细胞命运是至关重要的，以获得一个准确的理解细胞内Ca 2+信号的功能在凋亡诱导。
Stimulus-induced changes in the intracellular Ca2+concentration control cell fate decision, including apoptosis. However, the precise patterns of the cytosolic Ca2+signals that are associated with apoptotic induction remain unknown. We have developed a novel genetically encoded sensor of activated caspase-3 that can be applied in combination with a genetically encoded sensor of the Ca2+concentration and have established a dual imaging system that enables the imaging of both cytosolic Ca2+signals and caspase-3 activation, which is an indicator of apoptosis, in the same cell. Using this system, we identified differences in the cytosolic Ca2+signals of apoptotic and surviving DT40 B lymphocytes after B cell receptor (BCR) stimulation. In surviving cells, BCR stimulation evoked larger initial Ca2+spikes followed by a larger sustained elevation of the Ca2+concentration than those in apoptotic cells; BCR stimulation also resulted in repetitive transient Ca2+spikes, which were mediated by the influx of Ca2+from the extracellular space. Our results indicate that the observation of both Ca2+signals and cells fate in same cell is crucial to gain an accurate understanding of the function of intracellular Ca2+signals in apoptotic induction.