Visualisation of an nsPEF induced calcium wave using the genetically encoded calcium indicator GCaMP in U87 human glioblastoma cells

被引：9

作者：

Carr, Lynn ^{[1
]}

Bardet, Sylvia M. ^{[1
]}

Arnaud-Cormos, Delia ^{[1
]}

Leveque, Philippe ^{[1
]}

O'Connor, Rodney P. ^{[2
]}

机构：

[1] Univ Limoges, CNRS, XLIM, UMR 7252, F-87000 Limoges, France

[2] IMT Mines St Etienne, Ctr CMP, Dept Bioelect, F-13541 Gardanne, France

来源：

BIOELECTROCHEMISTRY | 2018年 / 119卷

关键词：

Genetically encoded calcium indicator; nsPEF; Plasma membrane calcium indicators; GCaMP; Calcium wave; PULSED ELECTRIC-FIELDS; INTRACELLULAR CA2+; CHROMAFFIN CELLS; MEMBRANE CA2+; NANOSECOND; RELEASE; VOLTAGE; ELECTROPERMEABILIZATION; TEMPERATURE; REVEALS;

D O I：

10.1016/j.bioelechem.2017.09.003

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Cytosolic, synthetic chemical calcium indicators are typically used to visualise the rapid increase in intracellular calcium ion concentration that follows nanosecond pulsed electric field (nsPEF) application. This study looks at the application of genetically encoded calcium indicators (GECIs) to investigate the spatiotemporal nature of nsPEF-induced calcium signals using fluorescent live cell imaging. Calcium responses to 44 kV/cm, 10 ns pulses were observed in U87-MG cells expressing either a plasma membrane targeted GECI (GCaMP5-G), or one cytosolically expressed (GCaMP6-S), and compared to the response of cells loaded with cytosolic or plasma membrane targeted chemical calcium indicators. Application of 100 pulses, to cells containing plasma membrane targeted indicators, revealed a wave of calcium across the cell initiating at the cathode side. A similar spatial wave was not observed with cytosolic indicators with mobile calcium buffering properties. The speed of the wave was related to pulse application frequency and it was not propagated by calcium induced calcium release. (C) 2017 Elsevier B.V. All rights reserved.

引用

页码：68 / 75

页数：8

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