A genetically encoded ratiometric calcium sensor enables quantitative measurement of the local calcium microdomain in the endoplasmic reticulum

Chen Luo; Huiyu Wang; Qi Liu; Wenting He; Lin Yuan; Pingyong Xu

doi:10.1007/s41048-019-0082-6

Volume 5 Issue 1

Turn off MathJax

Article Contents

Article Navigation > Biophysics Reports > 2019 > 5(1): 31-42

Chen Luo, Huiyu Wang, Qi Liu, Wenting He, Lin Yuan, Pingyong Xu. A genetically encoded ratiometric calcium sensor enables quantitative measurement of the local calcium microdomain in the endoplasmic reticulum. Biophysics Reports, 2019, 5(1): 31-42. doi: 10.1007/s41048-019-0082-6

Citation:

Chen Luo, Huiyu Wang, Qi Liu, Wenting He, Lin Yuan, Pingyong Xu. A genetically encoded ratiometric calcium sensor enables quantitative measurement of the local calcium microdomain in the endoplasmic reticulum. Biophysics Reports, 2019, 5(1): 31-42. doi: 10.1007/s41048-019-0082-6

Citation:

Chen Luo, Huiyu Wang, Qi Liu, Wenting He, Lin Yuan, Pingyong Xu. A genetically encoded ratiometric calcium sensor enables quantitative measurement of the local calcium microdomain in the endoplasmic reticulum. Biophysics Reports, 2019, 5(1): 31-42. doi: 10.1007/s41048-019-0082-6

PDF (3169 KB)

A genetically encoded ratiometric calcium sensor enables quantitative measurement of the local calcium microdomain in the endoplasmic reticulum

doi: 10.1007/s41048-019-0082-6

Chen Luo^1,2,
Huiyu Wang^1,2,
Qi Liu^1,2,
Wenting He¹,
Lin Yuan^{1
,
,},
Pingyong Xu^{1,2
,}

1 Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
2 College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China

Funds: This project was supported by the National Key R&D Program of China (2016YFA0501500), the National Natural Science Foundation of China (31421002, 31401174 and 21778069), and the Project of the Chinese Academy of Sciences (XDB08030203). We thank Mr. Junying Jia from the Institute of Biophysics, Chinese Academy of Science for his help in FACS data collection.

More Information

Corresponding author: Lin Yuan, Pingyong Xu
Received Date: 06 September 2018
Rev Recd Date: 15 November 2018
Publish Date: 28 February 2019

Abstract

Abstract

The local Ca²⁺ release from the heterogeneously distributed endoplasmic reticulum (ER) calcium store has a critical role in calcium homeostasis and cellular function. However, single fluorescent proteinbased ER calcium probes experience challenges in quantifying the ER calcium store in differing live cells, and intensity-based measurements make it difficult to detect local calcium microdomains in the ER. Here, we developed a genetically encoded ratiometric ER calcium indicator (GCEPIA1-SNAP_ER) that can detect the real-time ER calcium store and local calcium microdomains in live cells. GCEPIA1-SNAP_ER was located in the lumen of the ER and showed a linear, reversible and rapid response to changes in the ER calcium store. The GCEPIA1-SNAPER probe effectively monitored the depletion of the ER calcium store by TG or starvation treatment, and through its use we identified heterogeneously distributed calcium microdomains in the ER which were correlated with the distribution of STIM1 clusters upon ER calcium store depletion. Lastly, GCEPIA1-SNAP_ER can be used to detect the ER calcium store by highthroughput flow cytometry and confers the ability to study the function of calcium microdomains of the ER.
- Ratiometric calcium sensor,
- Local calcium microdomain,
- Endoplasmic reticulum

FullText(HTML)

References(21)

References

Akerboom J, Chen T-W, Wardill TJ, Tian L, Marvin JS, Mutlu S, Calderón NC, Esposti F, Borghuis BG, Sun XR (2012) Optimization of a GCaMP calcium indicator for neural activity imaging. J Neurosci 32:13819-13840

Carreras-Sureda A, Pihán P, Hetz C (2017) Calcium signaling at the endoplasmic reticulum:fine-tuning stress responses. Cell Calcium 70:24-31

Cho J-H, Swanson CJ, Chen J, Li A, Lippert LG, Boye SE, Rose K, Sivaramakrishnan S, Chuong C-M, Chow RH (2017) The GCaMP-R family of genetically encoded ratiometric calcium indicators. ACS Chem Biol 12:1066-1074

Clapham DE (2007) Calcium signaling. Cell 131:1047-1058

Cole NB (2013) Site-specific protein labeling with SNAP-tags. Curr Protoc Protein Sci. https://doi.org/10.1002/0471140864.ps3001s73

Ghislat G, Knecht E (2012) New Ca²⁺-dependent regulators of autophagosome maturation. Commun Integr Biol 5:308-311

Icha J, Weber M, Waters JC, Norden C (2017) Phototoxicity in live fluorescence microscopy, and how to avoid it. BioEssays 39:1700003

Koldenkova VP, Nagai T (2013) Genetically encoded Ca²⁺ indicators:properties and evaluation. Biochim Biophys Acta Mol Cell Res 1833:1787-1797

Liou J, Kim ML, Do Heo W, Jones JT, Myers JW, Ferrell JE Jr, Meyer T (2005) STIM is a Ca²⁺ sensor essential for Ca²⁺-storedepletion-triggered Ca²⁺ influx. Curr Biol 15:1235-1241

Mao T, O'Connor DH, Scheuss V, Nakai J, Svoboda K (2008) Characterization and subcellular targeting of GCaMP-type genetically-encoded calcium indicators. PLoS ONE 3:e1796

Park CY, Hoover PJ, Mullins FM, Bachhawat P, Covington ED, Raunser S, Walz T, Garcia KC, Dolmetsch RE, Lewis RS (2009) STIM1 clusters and activates CRAC channels via direct binding of a cytosolic domain to Orai1. Cell 136:876-890

Peinelt C, Vig M, Koomoa DL, Beck A, Nadler MJ, Koblan-Huberson M, Lis A, Fleig A, Penner R, Kinet J-P (2006) Amplification of CRAC current by STIM1 and CRACM1(Orai1). Nat Cell Biol 8:771

Prakriya M, Feske S, Gwack Y, Srikanth S, Rao A, Hogan PG (2006) Orai1 is an essential pore subunit of the CRAC channel. Nature 443:230

Rizzuto R, Pinton P, Carrington W, Fay FS, Fogarty KE, Lifshitz LM, Tuft RA, Pozzan T (1998) Close contacts with the endoplasmic reticulum as determinants of mitochondrial Ca²⁺ responses. Science 280:1763-1766

Shigetomi E, Kracun S, Khakh BS (2010) Monitoring astrocyte calcium microdomains with improved membrane targeted GCaMP reporters. Neuron Glia Biol 6:183-191

Stutzmann GE (2005) Calcium dysregulation, IP3 signaling, and Alzheimer's disease. The Neuroscientist 11:110-115

Suzuki J, Kanemaru K, Ishii K, Ohkura M, Okubo Y, Iino M (2014) Imaging intraorganellar Ca²⁺ at subcellular resolution using CEPIA. Nat Commun 5:4153

Tian L, Hires SA, Mao T, Huber D, Chiappe ME, Chalasani SH, Petreanu L, Akerboom J, McKinney SA, Schreiter ER (2009) Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators. Nat Methods 6:875

Wäldchen S, Lehmann J, Klein T, Van De Linde S, Sauer M (2015) Light-induced cell damage in live-cell super-resolution microscopy. Sci Rep 5:15348

Zhang SL, Yu Y, Roos J, Kozak JA, Deerinck TJ, Ellisman MH, Stauderman KA, Cahalan MD (2005) STIM1 is a Ca²⁺ sensor that activates CRAC channels and migrates from the Ca²⁺ store to the plasma membrane. Nature 437:902

Zhao Y, Araki S, Wu J, Teramoto T, Chang Y-F, Nakano M, Abdelfattah AS, Fujiwara M, Ishihara T, Nagai T (2011) An expanded palette of genetically encoded Ca²⁺ indicators. Science. https://doi.org/10.1126/science.1208592

Supplements(0)

Relative Articles

Cited By

Proportional views