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Simultaneous detection of L-glutamate and nitric oxide from adherently growing cells at known distance using disk shaped dual electrodes

Isik, Sonnur; Castillo Leon, Jaime LU ; Bloechl, Andrea; Csöregi, Elisabeth LU and Schuhmann, Wolfgang (2007) In Bioelectrochemistry 70(1). p.173-179
Abstract
An ex vivo system for simultaneous detection of nitric oxide (NO) and L-glutamate using integrated dual 250 gin platinum disk electrodes modified individually with suitable sensing chemistries has been developed. One of the sensors was coated with an electrocatalytic layer of Ni tetrasulfonate phthalocyanine tetrasodium salt (Ni-TSPe) covered by second layer of Nation, which stabilises on the one hand the primary oxidation product NO+ and prevents interferences from negatively charged compounds such as NO2-. For glutamate determination, the second electrode was modified with a crosslinked redox hydrogel consisting of Os complex modified poly(vinylimidazol), glutamate oxidase and peroxidase. A manual x-y-z micromanipulator on top of an... (More)
An ex vivo system for simultaneous detection of nitric oxide (NO) and L-glutamate using integrated dual 250 gin platinum disk electrodes modified individually with suitable sensing chemistries has been developed. One of the sensors was coated with an electrocatalytic layer of Ni tetrasulfonate phthalocyanine tetrasodium salt (Ni-TSPe) covered by second layer of Nation, which stabilises on the one hand the primary oxidation product NO+ and prevents interferences from negatively charged compounds such as NO2-. For glutamate determination, the second electrode was modified with a crosslinked redox hydrogel consisting of Os complex modified poly(vinylimidazol), glutamate oxidase and peroxidase. A manual x-y-z micromanipulator on top of an inverted optical microscope was used to position the dual electrode sensor at a defined distance of 5 mu m from a cell population under visual control. C6 glioma cells were stimulated simultaneously with bradykinin or VEGF to release NO while KCl was used to invoke glutamate release. For evaluation of the glutamate sensors, in some experiments HN10 cells were used. To investigate the sensitivity and reliability of the system, several drugs were applied to the cells, e.g. Ca2+-channel inhibitors for testing Ca2+-dependence of the release of NO and glutamate, rotenone for inducing oxidative stress and glutamate antagonists for analysing glutamate release. With these drugs the NO and glutamate release was modulated in a similar way then expected from previously described systems or even in-vivo measurements. We therefore conclude that our system is suitable to analyse stress-induced mechanisms in cell lines. (c) 2006 Elsevier B.V All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
antagonists, glutamate, nitric oxide, dual electrode, simultaneous detection, drugs
in
Bioelectrochemistry
volume
70
issue
1
pages
173 - 179
publisher
Elsevier
external identifiers
  • wos:000243708600029
  • scopus:33845679296
ISSN
1878-562X
DOI
10.1016/j.bioelechem.2006.03.037
language
English
LU publication?
yes
id
64e97671-6f81-4c91-a001-fd92f920b715 (old id 676425)
date added to LUP
2007-12-13 09:20:09
date last changed
2017-07-30 03:38:35
@article{64e97671-6f81-4c91-a001-fd92f920b715,
  abstract     = {An ex vivo system for simultaneous detection of nitric oxide (NO) and L-glutamate using integrated dual 250 gin platinum disk electrodes modified individually with suitable sensing chemistries has been developed. One of the sensors was coated with an electrocatalytic layer of Ni tetrasulfonate phthalocyanine tetrasodium salt (Ni-TSPe) covered by second layer of Nation, which stabilises on the one hand the primary oxidation product NO+ and prevents interferences from negatively charged compounds such as NO2-. For glutamate determination, the second electrode was modified with a crosslinked redox hydrogel consisting of Os complex modified poly(vinylimidazol), glutamate oxidase and peroxidase. A manual x-y-z micromanipulator on top of an inverted optical microscope was used to position the dual electrode sensor at a defined distance of 5 mu m from a cell population under visual control. C6 glioma cells were stimulated simultaneously with bradykinin or VEGF to release NO while KCl was used to invoke glutamate release. For evaluation of the glutamate sensors, in some experiments HN10 cells were used. To investigate the sensitivity and reliability of the system, several drugs were applied to the cells, e.g. Ca2+-channel inhibitors for testing Ca2+-dependence of the release of NO and glutamate, rotenone for inducing oxidative stress and glutamate antagonists for analysing glutamate release. With these drugs the NO and glutamate release was modulated in a similar way then expected from previously described systems or even in-vivo measurements. We therefore conclude that our system is suitable to analyse stress-induced mechanisms in cell lines. (c) 2006 Elsevier B.V All rights reserved.},
  author       = {Isik, Sonnur and Castillo Leon, Jaime and Bloechl, Andrea and Csöregi, Elisabeth and Schuhmann, Wolfgang},
  issn         = {1878-562X},
  keyword      = {antagonists,glutamate,nitric oxide,dual electrode,simultaneous detection,drugs},
  language     = {eng},
  number       = {1},
  pages        = {173--179},
  publisher    = {Elsevier},
  series       = {Bioelectrochemistry},
  title        = {Simultaneous detection of L-glutamate and nitric oxide from adherently growing cells at known distance using disk shaped dual electrodes},
  url          = {http://dx.doi.org/10.1016/j.bioelechem.2006.03.037},
  volume       = {70},
  year         = {2007},
}