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Measurements of Dynamic [Cl‐]i responses in stably C1C2 expressed cells with quantum dots(QD) chloride nanobiosensor, Cl‐QD
Author(s) -
Mao Hua,
Wang Yuchi,
Wang Ling,
Wong Lid B
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.lb760
Subject(s) - dids , chloride channel , chloride , glibenclamide , chemistry , transfection , biophysics , cell culture , microbiology and biotechnology , biochemistry , biology , membrane , gene , endocrinology , genetics , organic chemistry , diabetes mellitus
C1C‐2 channel is one of the drug targets for the treatments of chloride channelopathies and imbalanced fluid secretions. Herein we describe the results of the [Cl − ]i in a stably expressed ClC‐2 transfected HEK293F and T84 human cell lines measured by Cl‐ QD™. We have evaluated the role of C1C‐2 on [Cl − ]i homeostasis with different putative chloride channel inhibitors and activators in both HEK293F/HEK293F C1C2 and T84/T84 C1C2 . Cl‐QDs were loaded into the cells using a cell‐penetrating TAT peptide protocol. [Cl − ]i responses were measured using a photon counting fluorescence microscope. Five experiments were performed per condition. All four tested chloride channel inhibitors, DIDS, DCPIB, DPC and glibenclamide, caused dose‐dependent increases of [Cl − ]i in C1C‐2 expressed cells compared to their respective wild types. Conversely, both tested chloride channel activators, lubriprostone and genistein, caused a lesser decrease of [Cl − ]i in the C1C‐2 expressed cells compared to their respective wild types. These data demonstrate that cells stably expressed with ClC‐2 channels enhanced the responses of many putative chloride inhibitors. These data support the hypothesis that C1C‐2 channels could be a potential drug target for the treatment of diarrhea, an alternate option to the conventional cAMP‐ and Ca‐dependent chloride pathways. (Supported by NIDDK R44‐DK084600)