Imaging intraorganellar Ca2+ at subcellular resolution using CEPIA

Junji Suzuki, Kazunori Kanemaru, Kuniaki Ishii, Masamichi Ohkura, Yohei Okubo and Masamitsu Iino ()
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Junji Suzuki: Graduate School of Medicine, The University of Tokyo
Kazunori Kanemaru: Graduate School of Medicine, The University of Tokyo
Kuniaki Ishii: School of Medicine, Yamagata University
Masamichi Ohkura: Brain Science Institute, Saitama University
Yohei Okubo: Graduate School of Medicine, The University of Tokyo
Masamitsu Iino: Graduate School of Medicine, The University of Tokyo

Nature Communications, 2014, vol. 5, issue 1, 1-13

Abstract: Abstract The endoplasmic reticulum (ER) and mitochondria accumulate Ca2+ within their lumens to regulate numerous cell functions. However, determining the dynamics of intraorganellar Ca2+ has proven to be difficult. Here we describe a family of genetically encoded Ca2+ indicators, named calcium-measuring organelle-entrapped protein indicators (CEPIA), which can be utilized for intraorganellar Ca2+ imaging. CEPIA, which emit green, red or blue/green fluorescence, are engineered to bind Ca2+ at intraorganellar Ca2+ concentrations. They can be targeted to different organelles and may be used alongside other fluorescent molecular markers, expanding the range of cell functions that can be simultaneously analysed. The spatiotemporal resolution of CEPIA makes it possible to resolve Ca2+ import into individual mitochondria while simultaneously measuring ER and cytosolic Ca2+. We have used these imaging capabilities to reveal differential Ca2+ handling in individual mitochondria. CEPIA imaging is a useful new tool to further the understanding of organellar functions.

Date: 2014
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DOI: 10.1038/ncomms5153

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