Genetically encoded chloride indicator with improved sensitivity.

Abstract : Chloride (Cl) is the most abundant physiological anion. Abnormalities in Cl regulation are instrumental in the development of several important diseases including motor disorders and epilepsy. Because of difficulties in the spectroscopic measurement of Cl in live tissues there is little knowledge available regarding the mechanisms of regulation of intracellular Cl concentration. Several years ago, a CFP-YFP based ratiometric Cl indicator (Clomeleon) was introduced [Kuner, T., Augustine, G.J. A genetically encoded ratiometric indicator for chloride: capturing chloride transients in cultured hippocampal neurons. Neuron 2000; 27: 447-59]. This construct with relatively low sensitivity to Cl (K(app) approximately 160 mM) allows ratiometric monitoring of Cl using fluorescence emission ratio. Here, we propose a new CFP-YFP-based construct (Cl-sensor) with relatively high sensitivity to Cl (K(app) approximately 30 mM) due to triple YFP mutant. The construct also exhibits good pH sensitivity with pK(alpha) ranging from 7.1 to 8.0 pH units at different Cl concentrations. Using Cl-sensor we determined non-invasively the distribution of [Cl](i) in cultured CHO cells, in neurons of primary hippocampal cultures and in photoreceptors of rat retina. This genetically encoded indicator offers a means for monitoring Cl and pH under different physiological conditions and high-throughput screening of pharmacological agents.
Type de document :
Article dans une revue
Journal of Neuroscience Methods, Elsevier, 2008, 170 (1), pp.67-76. 〈10.1016/j.jneumeth.2007.12.016〉
Liste complète des métadonnées

Littérature citée [62 références]  Voir  Masquer  Télécharger

http://www.hal.inserm.fr/inserm-00483494
Contributeur : Roman Tyzio <>
Soumis le : vendredi 14 mai 2010 - 13:27:03
Dernière modification le : mardi 30 janvier 2018 - 01:00:43
Document(s) archivé(s) le : mercredi 30 novembre 2016 - 22:59:09

Fichier

 Accès restreint
Fichier visible le : jamais

Connectez-vous pour demander l'accès au fichier

Identifiants

Collections

Citation

Olga Markova, Marat Mukhtarov, Eleonore Real, Yves Jacob, Piotr Bregestovski. Genetically encoded chloride indicator with improved sensitivity.. Journal of Neuroscience Methods, Elsevier, 2008, 170 (1), pp.67-76. 〈10.1016/j.jneumeth.2007.12.016〉. 〈inserm-00483494〉

Partager

Métriques

Consultations de la notice

103