Imaging elemental events of store-operated Ca2+ entry in invading cancer cells with plasmalemmal targeted sensors

Fujian Lu, Jianwei Sun, Qiaoxia Zheng, Jinghang Li, Yuanzhao Hu, Peng Yu, Huifang He, Yan Zhao, Xianhua Wang, Shengyu Yang, Heping Cheng

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

STIM1- and Orai1-mediated store-operated Ca2+ entry (SOCE) constitutes the major Ca2+ influx in almost all electrically nonexcitable cells. However, little is known about the spatiotemporal organization at the elementary level. Here, we developed Orai1-tethered or palmitoylated biosensor GCaMP6f to report subplasmalemmal Ca 2+ signals. We visualized spontaneous discrete and long-lasting transients (Ca 2+ glows') arising from STIM1-Orai1 in invading melanoma cells. Ca 2+ glows occurred preferentially in single invadopodia and at sites near the cell periphery under resting conditions. Re-addition of external Ca2+ after store depletion elicited spatially synchronous Ca2+ glows, followed by high-rate discharge of asynchronous local events. Knockout of STIM1 or expression of the dominant-negative Orai1-E106A mutant markedly decreased Ca2+ glow frequency, diminished global SOCE and attenuated invadopodial formation. Functionally, invadopodial Ca2+ glows provided high Ca2+ microdomains to locally activate Ca2+ /calmodulin-dependent Pyk2 (also known as PTK2B), which initiates the SOCE-Pyk2-Src signaling cascade required for invasion.Overall, the discovery of elemental Ca2+ signals of SOCE not only unveils a previously unappreciated gating mode of STIM1-Orai1 channels in situ, but also underscores a critical role of the spatiotemporal dynamics of SOCE in orchestrating complex cell behaviors such as invasion. This article has an associated First Person interview with the first author of the paper.

Original languageEnglish (US)
Article number224923
JournalJournal of Cell Science
Volume132
Issue number6
DOIs
StatePublished - Mar 1 2019

Fingerprint

Neoplasms
Biosensing Techniques
Calmodulin
Melanoma
Interviews
Podosomes

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Lu, Fujian ; Sun, Jianwei ; Zheng, Qiaoxia ; Li, Jinghang ; Hu, Yuanzhao ; Yu, Peng ; He, Huifang ; Zhao, Yan ; Wang, Xianhua ; Yang, Shengyu ; Cheng, Heping. / Imaging elemental events of store-operated Ca2+ entry in invading cancer cells with plasmalemmal targeted sensors. In: Journal of Cell Science. 2019 ; Vol. 132, No. 6.
@article{b01dac88395a4053937e1fee9e023a23,
title = "Imaging elemental events of store-operated Ca2+ entry in invading cancer cells with plasmalemmal targeted sensors",
abstract = "STIM1- and Orai1-mediated store-operated Ca2+ entry (SOCE) constitutes the major Ca2+ influx in almost all electrically nonexcitable cells. However, little is known about the spatiotemporal organization at the elementary level. Here, we developed Orai1-tethered or palmitoylated biosensor GCaMP6f to report subplasmalemmal Ca 2+ signals. We visualized spontaneous discrete and long-lasting transients (Ca 2+ glows') arising from STIM1-Orai1 in invading melanoma cells. Ca 2+ glows occurred preferentially in single invadopodia and at sites near the cell periphery under resting conditions. Re-addition of external Ca2+ after store depletion elicited spatially synchronous Ca2+ glows, followed by high-rate discharge of asynchronous local events. Knockout of STIM1 or expression of the dominant-negative Orai1-E106A mutant markedly decreased Ca2+ glow frequency, diminished global SOCE and attenuated invadopodial formation. Functionally, invadopodial Ca2+ glows provided high Ca2+ microdomains to locally activate Ca2+ /calmodulin-dependent Pyk2 (also known as PTK2B), which initiates the SOCE-Pyk2-Src signaling cascade required for invasion.Overall, the discovery of elemental Ca2+ signals of SOCE not only unveils a previously unappreciated gating mode of STIM1-Orai1 channels in situ, but also underscores a critical role of the spatiotemporal dynamics of SOCE in orchestrating complex cell behaviors such as invasion. This article has an associated First Person interview with the first author of the paper.",
author = "Fujian Lu and Jianwei Sun and Qiaoxia Zheng and Jinghang Li and Yuanzhao Hu and Peng Yu and Huifang He and Yan Zhao and Xianhua Wang and Shengyu Yang and Heping Cheng",
year = "2019",
month = "3",
day = "1",
doi = "10.1242/jcs.224923",
language = "English (US)",
volume = "132",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "6",

}

Imaging elemental events of store-operated Ca2+ entry in invading cancer cells with plasmalemmal targeted sensors. / Lu, Fujian; Sun, Jianwei; Zheng, Qiaoxia; Li, Jinghang; Hu, Yuanzhao; Yu, Peng; He, Huifang; Zhao, Yan; Wang, Xianhua; Yang, Shengyu; Cheng, Heping.

In: Journal of Cell Science, Vol. 132, No. 6, 224923, 01.03.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Imaging elemental events of store-operated Ca2+ entry in invading cancer cells with plasmalemmal targeted sensors

AU - Lu, Fujian

AU - Sun, Jianwei

AU - Zheng, Qiaoxia

AU - Li, Jinghang

AU - Hu, Yuanzhao

AU - Yu, Peng

AU - He, Huifang

AU - Zhao, Yan

AU - Wang, Xianhua

AU - Yang, Shengyu

AU - Cheng, Heping

PY - 2019/3/1

Y1 - 2019/3/1

N2 - STIM1- and Orai1-mediated store-operated Ca2+ entry (SOCE) constitutes the major Ca2+ influx in almost all electrically nonexcitable cells. However, little is known about the spatiotemporal organization at the elementary level. Here, we developed Orai1-tethered or palmitoylated biosensor GCaMP6f to report subplasmalemmal Ca 2+ signals. We visualized spontaneous discrete and long-lasting transients (Ca 2+ glows') arising from STIM1-Orai1 in invading melanoma cells. Ca 2+ glows occurred preferentially in single invadopodia and at sites near the cell periphery under resting conditions. Re-addition of external Ca2+ after store depletion elicited spatially synchronous Ca2+ glows, followed by high-rate discharge of asynchronous local events. Knockout of STIM1 or expression of the dominant-negative Orai1-E106A mutant markedly decreased Ca2+ glow frequency, diminished global SOCE and attenuated invadopodial formation. Functionally, invadopodial Ca2+ glows provided high Ca2+ microdomains to locally activate Ca2+ /calmodulin-dependent Pyk2 (also known as PTK2B), which initiates the SOCE-Pyk2-Src signaling cascade required for invasion.Overall, the discovery of elemental Ca2+ signals of SOCE not only unveils a previously unappreciated gating mode of STIM1-Orai1 channels in situ, but also underscores a critical role of the spatiotemporal dynamics of SOCE in orchestrating complex cell behaviors such as invasion. This article has an associated First Person interview with the first author of the paper.

AB - STIM1- and Orai1-mediated store-operated Ca2+ entry (SOCE) constitutes the major Ca2+ influx in almost all electrically nonexcitable cells. However, little is known about the spatiotemporal organization at the elementary level. Here, we developed Orai1-tethered or palmitoylated biosensor GCaMP6f to report subplasmalemmal Ca 2+ signals. We visualized spontaneous discrete and long-lasting transients (Ca 2+ glows') arising from STIM1-Orai1 in invading melanoma cells. Ca 2+ glows occurred preferentially in single invadopodia and at sites near the cell periphery under resting conditions. Re-addition of external Ca2+ after store depletion elicited spatially synchronous Ca2+ glows, followed by high-rate discharge of asynchronous local events. Knockout of STIM1 or expression of the dominant-negative Orai1-E106A mutant markedly decreased Ca2+ glow frequency, diminished global SOCE and attenuated invadopodial formation. Functionally, invadopodial Ca2+ glows provided high Ca2+ microdomains to locally activate Ca2+ /calmodulin-dependent Pyk2 (also known as PTK2B), which initiates the SOCE-Pyk2-Src signaling cascade required for invasion.Overall, the discovery of elemental Ca2+ signals of SOCE not only unveils a previously unappreciated gating mode of STIM1-Orai1 channels in situ, but also underscores a critical role of the spatiotemporal dynamics of SOCE in orchestrating complex cell behaviors such as invasion. This article has an associated First Person interview with the first author of the paper.

UR - http://www.scopus.com/inward/record.url?scp=85063712131&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85063712131&partnerID=8YFLogxK

U2 - 10.1242/jcs.224923

DO - 10.1242/jcs.224923

M3 - Article

C2 - 30814332

AN - SCOPUS:85063712131

VL - 132

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 6

M1 - 224923

ER -