Genetically-encoded calcium indicator for detecting the dynamics of drug-induced nuclear calcium signals in real time

doi:10.3969/j.issn.1006-7795.2018.03.013

Journal of Capital Medical University ›› 2018, Vol. 39 ›› Issue (3): 378-384.doi: 10.3969/j.issn.1006-7795.2018.03.013

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Genetically-encoded calcium indicator for detecting the dynamics of drug-induced nuclear calcium signals in real time

Gong Mingtao¹, Zhang Chenguang^1,2, Ding Wei^1,2

1. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;
2. Beijing Key Laboratory for Researches in Cancer Invasion and Metastasis, Cancer Institute of Capital Medical University, Beijing 100069, China

Received:2017-05-15 Online:2018-05-21 Published:2018-06-11
Supported by:
This study was supported by National Natural Science Foundation of China (81372284).

Abstract

Abstract: Objective To explore the application of genetically-encoded calcium indicator for detecting the dynamics of drug-induced nuclear calcium signals in real time.Methods We fused tdTomato with red fluorescence as an intramolecular reference to a most commonly used GeCI variant GCaMP6s, and tagged the molecule with the nucleoplasmin nuclear localization sequence (NLS) into an eukaryotic expression cassette. By transfecting HeLa or U87MG cells, and imaged with a live confocal microscope, we were able to quantitatively monitor the dynamic changes of nuclear calcium signals in real time.Results We found that the ability of Paxilline to increase nuclear calcium concentrations varied among different cell types. We also found that various chemical, including artemisinin and several cell stress inducing reagent, were able to elevate nuclear calcium level in a relatively persistent manner.Conclusion Different drugs can increase calcium concentration in non-excitable cells.

Key words: calcium indicator, live fluorescent imaging, nuclear signal, artemisinin, high-content

CLC Number:

Gong Mingtao, Zhang Chenguang, Ding Wei. Genetically-encoded calcium indicator for detecting the dynamics of drug-induced nuclear calcium signals in real time[J]. Journal of Capital Medical University, 2018, 39(3): 378-384.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2018.03.013

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Genetically-encoded calcium indicator for detecting the dynamics of drug-induced nuclear calcium signals in real time

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