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

Authors
  • Carr, Lynn1
  • Bardet, Sylvia M1
  • Arnaud-Cormos, Delia1
  • Leveque, Philippe1
  • O'Connor, Rodney P2
  • 1 Univ. Limoges, CNRS, XLIM, UMR 7252, F-87000 Limoges, France. , (France)
  • 2 IMT Mines Saint-Etienne, Centre CMP, Department of Bioelectronics, F - 13541 Gardanne, France. Electronic address: [email protected] , (France)
Type
Published Article
Journal
Bioelectrochemistry (Amsterdam, Netherlands)
Publication Date
Feb 01, 2018
Volume
119
Pages
68–75
Identifiers
DOI: 10.1016/j.bioelechem.2017.09.003
PMID: 28917183
Source
Medline
Keywords
License
Unknown

Abstract

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 44kV/cm, 10ns 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.

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