Investigation into the difference in mitochondrial-cytosolic calcium coupling between adult cardiomyocyte and hiPSC-CM using a novel multifunctional genetic probe.
Investigation into the difference in mitochondrial-cytosolic calcium coupling between adult cardiomyocyte and hiPSC-CM using a novel multifunctional genetic probe.
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用新型多功能遗传探针研究成年心肌细胞与HiPSC-CM线粒体-胞浆钙偶联的差异。
DOI：
10.1007/s00424-021-02524-3
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发表时间：
2021-03
期刊：
影响因子：
--
通讯作者：
Zhou L
中科院分区：
文献类型：
--
作者：
Ernst P;Chen K;Tang Y;Kim S;Guan J;He J;Xie M;Zhang JJ;Liu XM;Zhou L
在生理和病理条件下，Ca2+循环在调节心肌细胞（CM）功能中起关键作用。线粒体参与成人心肌细胞（ACMs）的Ca2+处理。然而，关于它们在人诱导多能干细胞衍生心肌细胞（hiPSC-CMs）中Ca2+动力学调节中的作用知之甚少。在本研究中，我们开发了一种多功能的遗传编码Ca2+探针，能够同时实时测量细胞质和线粒体Ca2+。利用这种新型探针，我们测定并比较了hiPSC-CMs和ACMs中线粒体Ca2+活性及其与细胞质Ca2+动力学的耦合。我们的数据显示，虽然ACMs在线粒体Ca2+中显示出与细胞质Ca2+同步的高度协调的节奏反应，但hiPSC-CMs在线粒体Ca2+活性中显示出高度的细胞范围变异性，与细胞质Ca2+协调不良。然后我们发现，与ACM相比，hiPSC-CM中的线粒体-肌浆网（SR）系结以及线粒体间网络连接不发达，这可能是细胞质和线粒体Ca2+动力学之间观察到的时空解耦的基础。最后，我们发现，mitofusin-2（一种连接线粒体和SR的蛋白质）的敲低导致ACMs中细胞质-线粒体Ca2+偶联减少，尽管与hiPSC-CMs相比程度较低，这表明Mfn2是改善hiPSC-CMs中线粒体-细胞质Ca2+偶联的潜在工程靶点。本研究将促进我们对线粒体在CMs中Ca2+处理和循环中的作用的理解，并指导hiPSC-CMs治疗损伤心脏的发展。
Ca2+ cycling plays a critical role in regulating cardiomyocyte (CM) function under both physiological and pathological conditions. Mitochondria have been implicated in Ca2+ handling in adult cardiomyocytes (ACMs). However, little is known about their role in the regulation of Ca2+ dynamics in human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs). In the present study, we developed a multifunctional genetically-encoded Ca2+ probe capable of simultaneously measuring cytosolic and mitochondrial Ca2+ in real time. Using this novel probe, we determined and compared mitochondrial Ca2+ activity and the coupling with cytosolic Ca2+ dynamics in hiPSC-CMs and ACMs. Our data showed that while ACMs displayed a highly coordinated beat-by-beat response in mitochondrial Ca2+ in sync with cytosolic Ca2+,whereas hiPSC-CMs showed high cell-wide variability in mitochondrial Ca2+ activity that is poorly coordinated with cytosolic Ca2+. We then revealed that mitochondrial-sarcoplasmic reticulum (SR) tethering, as well as the inter-mitochondrial network connection, are underdeveloped in hiPSC-CM compared to ACM, which may underlie the observed spatiotemporal decoupling between cytosolic and mitochondrial Ca2+ dynamics. Finally, we showed that knockdown of mitofusin-2 (Mfn2), a protein tethering mitochondria and SR, led to reduced cytosolic-mitochondrial Ca2+ coupling in ACMs, albeit to a lesser degree compared to hiPSC-CMs, suggesting that Mfn2 is a potential engineering target for improving mitochondrial-cytosolic Ca2+ coupling in hiPSC-CMs. The present study will advance our understanding of the role of mitochondria in Ca2+ handling and cycling in CMs, and guide the development of hiPSC-CMs for healing injured hearts.
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影响因子：
5
作者：
Leonard A;Bertero A;Powers JD;Beussman KM;Bhandari S;Regnier M;Murry CE;Sniadecki NJ
通讯作者：
Sniadecki NJ
影响因子：
--
作者：
Guo L;Eldridge S;Furniss M;Mussio J;Davis M
通讯作者：
Davis M
影响因子：
10.8
作者：
Goh, Kah Yong;Qu, Jing;Zhou, Lufang
通讯作者：
Zhou, Lufang
DOI：
10.1007/978-1-61779-382-0_14
发表时间：
2012-01-01
期刊：
MITOCHONDRIAL BIOENERGETICS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Davidson, Sean M.;Duchen, Michael R.
通讯作者：
Duchen, Michael R.
影响因子：
4.6
作者：
Cortes, David;McTiernan, Christopher D.;Alarcon, Emilio, I
通讯作者：
Alarcon, Emilio, I