Integrated microsystem for non-invasive electrophysiological measurements on Xenopus oocytes

Détails

ID Serval
serval:BIB_A3846B617EB3
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Integrated microsystem for non-invasive electrophysiological measurements on Xenopus oocytes
Périodique
Biosensors and Bioelectronics
Auteur⸱e⸱s
Dahan  E., Bize  V., Lehnert  T., Horisberger  J. D., Gijs  M. A.
ISSN
0956-5663 (Print)
Statut éditorial
Publié
Date de publication
06/2007
Volume
22
Numéro
12
Pages
3196-202
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jun 15
Résumé
We propose a new non-invasive integrated microsystem for electrophysiological measurements on Xenopus laevis oocytes. Xenopus oocyte is a well-known expression system for various kinds of ion channels, that are potential tools in drug screening. In the traditional "Two Electrode Voltage Clamp" (TEVC) method, delicate micromanipulation is required to impale an oocyte with two microelectrodes. In our system, a non-invasive electrical access to the cytoplasm is provided by permeabilizing the cell membrane with an ionophore (e.g. nystatin). Unlike the classical patch-clamp or "macropatch" techniques, this method does not require removal of the vitelline membrane. Cell handling is significantly simplified, resulting in more robust recordings with increased throughput. Moreover, because only part of the oocyte surface is exposed to reagents, the required volume of reagent solutions could be reduced by an order of magnitude compared to the TEVC method. The fabrication process for this disposable microchip, based on poly-dimethylsiloxane (PDMS) molding and glass/PDMS bonding, is cost-efficient and simple. We tested this new microdevice by recording currents in oocytes expressing the human Epithelial Sodium Channel (hENaC) for membrane potentials between -100 and +50 mV. We recorded benzamil-sensitive currents with a large signal-to-noise ratio and we also obtained a benzamil concentration-inhibition curve displaying an inhibition constant IC(50) of about 50 nM, comparable to previously published values obtained with the TEVC technique.
Mots-clé
Amiloride/analogs & derivatives/pharmacology Animals Dimethylpolysiloxanes/chemistry Dose-Response Relationship, Drug Electrodes Epithelial Sodium Channel/drug effects/physiology Female Humans Membrane Potentials Microchip Analytical Procedures/*instrumentation Oocytes/*physiology Patch-Clamp Techniques/*instrumentation Silicones/chemistry Xenopus
Pubmed
Web of science
Création de la notice
24/01/2008 12:38
Dernière modification de la notice
20/08/2019 15:09
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