Excitation of mitochondria-Endoplasmic reticulum Ca2+ coupling by femtosecond-Laser photostimulation
Excitation of mitochondria-Endoplasmic reticulum Ca2+ coupling by femtosecond-Laser photostimulation
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DOI：
10.1109/jphot.2020.3004837
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发表时间：
2020-06
影响因子：
2.4
通讯作者：
Fan Shi;Shaoyang Wang;Yujie Zhu;Hao He
中科院分区：
文献类型：
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作者：
Fan Shi;Shaoyang Wang;Yujie Zhu;Hao He
线粒体位于内质网（ER）附近，与ER合作编码细胞内钙信号，参与细胞的过程和功能。然而，线粒体和ER之间的钙耦合（mito-ER）仍然难以捉摸，由于难以精确激发局部mito-ER钙耦合，对其他细胞器的干扰很小。为了研究线粒体-内质网钙偶联动力学，我们报道了一种非侵入性的飞秒激光光刺激方法，在单光子水平上激发线粒体-内质网钙偶联。我们建立了一个显微镜和光刺激系统，同时刺激和观察单个突触在真实的时间连续，通过一个长期的线粒体-ER钙耦合观察后，一个单一的飞秒激光短闪光刺激的管状结构的目标突触在预先选定的活细胞。线粒体-内质网钙偶联中胞浆钙离子参与，但线粒体钠钙交换体和线粒体-内质网距离对线粒体-内质网钙偶联的影响很小，线粒体-内质网钙偶联的动力学与生化药物激发的线粒体-内质网钙偶联动力学完全不同。本研究为线粒体及线粒体-内质网钙偶联的研究提供了一种在单个细胞器水平上激发钙动力学的精确、无创的方法。
Mitochondria locate close to endoplasmic reticulum (ER) and cooperate with ER to encode intracellular calcium signaling for cell processes and functions. However, the calcium coupling between mitochondria and ER (mito-ER) remains elusive due to the difficulty of precise excitation of localized mito-ER calcium coupling with little interference to other organelles. To study the mito-ER calcium coupling dynamics, we report a noninvasive method of photostimulation by femtosecond laser to excite the mito-ER calcium coupling at single-mitochondrion level. We establish a microscopy and photostimulation system to simultaneously stimulate and observe single mitochondrion continuously in real time, by which a long-term mito-ER calcium coupling is observed after a single short flash of femtosecond-laser stimulation to a tubular structure of a target mitochondrion in a pre-selected live cell. Cytosolic calcium is involved in the mito-ER calcium coupling but the mitochondrial sodium calcium exchanger and the mito-ER distance contribute little to it. Single mito-ER calcium coupling exhibits different dynamics from it excited by biochemical drugs globally. This study provides a precise and noninvasive method to excite calcium dynamics at single organelle level for the study of mitochondria and mito-ER calcium coupling.