Visible light excited ratiometric-GECIs for long-term in-cellulo monitoring of calcium signals

Jia Li; Liuqing Wang; Yilan Chen; Yang Yang; Jin Liu; Kai Liu; Yi-Tsang Lee; Ningjia He; Yubin Zhou; Youjun Wang

doi:10.1016/j.ceca.2020.102165

Visible light excited ratiometric-GECIs for long-term in-cellulo monitoring of calcium signals

Cell Calcium. 2020 May:87:102165. doi: 10.1016/j.ceca.2020.102165. Epub 2020 Jan 23.

Authors

Jia Li¹, Liuqing Wang¹, Yilan Chen¹, Yang Yang², Jin Liu¹, Kai Liu¹, Yi-Tsang Lee³, Ningjia He⁴, Yubin Zhou³, Youjun Wang⁵

Affiliations

¹ Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, China.
² School of Civil Engineering and Transportation, South China University of Technology, Guangdong, 510000, China.
³ Institute of Biosciences and Technology, Texas A&M University, Houston, TX, 77030, USA.
⁴ State Key Laboratory of Silkworm Genome Biology, Southwest University, Beibei, Chongqing, 400715, China.
⁵ Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, China; Key Laboratory of Cell Proliferation and Regulation Biology, Ministry of Education, Institute of Cell Biology, College of Life Sciences, Beijing Normal University, Beijing, 100875, China. Electronic address: wyoujun@bnu.edu.cn.

PMID: 32004817
DOI: 10.1016/j.ceca.2020.102165

Abstract

Genetically Encoded Calcium Indicators (GECIs) are powerful molecular tools for monitoring calcium (Ca²⁺) signaling in the cytosol and organellar compartments. However, currently available ratiometric indicators that allow measurements of resting Ca²⁺ levels have limitations in long-term Ca²⁺ imaging. They either are ultraviolet (UV)-excited ones with strong photo-toxicity, or have poor performance. To overcome this hurdle, we developed a set of visible light excited ratiometric-GECIs (VR-GECIs) based on existing mono-colored GECIs. With performance comparable to their corresponding mono-color prototypes, this set of VR-GECIs enables long-term measurements of intra-cellular or intra-organellar Ca²⁺ signals. Using these VR-GECIs together with a newly developed off-line analysis tool, we achieved long-term measurements of Ca²⁺ homeostasis of moving or dividing cells. Our tools may find broad applications in decoding Ca²⁺-modulated physiological or pathological processes.

Keywords: Calcium; ER; GECI; Homeostasis; Long-term; Mitochondria; Mitosis; Ratiometric.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Calcium / metabolism*
Calcium Signaling / radiation effects*
Cell Shape
Endoplasmic Reticulum / metabolism
Fluorescence
HEK293 Cells
HeLa Cells
Humans
Light*
Mitosis

Substances

Calcium