Title:
DETECTION OF TRANSMEMBRANE POTENTIALS BY OPTICAL METHODS
Document Type and Number:
WIPO Patent Application WO2001042211
Kind Code:
A3
Abstract:
Methods and compositions are provided for detecting changes in membrane potential in membranes biological systems. In one aspect, the method comprises: a) providing a living cell with a first reagent comprising a charged hydrophobic molecule which is typically a fluorescence resonance energy transfer (FRET) acceptor or donor, or is a quencher and is capable of redistributing within the membrane of a biological membrane in response to changes in the potential across the membrane; b) providing the cell with a second reagent that can label the first face or the second face of a biological membrane within the cell; c) detecting light emission from the first reagent or the second reagent. One aspect of this method involves monitoring membrane potential changes in subcellular organelle membranes in a living cell. Another aspect of the invention is the use of certain embodiments of the method for the screening of test chemicals for activity to modulate the activity of a target ion channel. Another aspect of the present invention is a transgenic organism comprising a first reagent that comprises a charged hydrophobic fluorescent molecule, and a second reagent comprising a bioluminescent or naturally fluorescent protein.
Inventors:
TSIEN ROGER Y
GONZALEZ JESUS E III
GONZALEZ JESUS E III
Application Number:
PCT/US2000/033739
Publication Date:
January 17, 2002
Filing Date:
December 12, 2000
Export Citation:
Assignee:
UNIV CALIFORNIA (US)
International Classes:
C07D209/08; C07D213/53; C07D213/74; C07D215/22; C07D215/227; C07D215/48; C07D215/50; C07D215/54; C07D239/60; C07D239/62; C07D239/66; C07D239/80; C07D241/44; C07D265/36; C07D279/16; G01N21/78; C07D307/14; C07D311/08; C07D311/16; C07D335/06; C07D487/04; C07F5/00; C07F5/02; C07F5/05; C07F9/10; C07F9/572; C07F9/6512; C09B23/02; C09K11/06; C12N5/10; C12N15/09; C12Q1/02; C12Q1/66; G01N21/64; G01N21/76; G01N27/416; G01N33/15; G01N33/483; G01N33/50; G01N33/52; G01N33/542; G01N33/58; G01N33/68; G01N33/92; (IPC1-7): C07D209/08; C07D213/74; C07D215/22; C07D215/48; C07D215/50; C07D215/54; C07D239/60; C07D311/08; C07D311/16; C07D335/06; C07F5/00; C07F5/02; C07K14/435; C09K11/06; C12N13/00
Domestic Patent References:
| WO1996041166A2 | 1996-12-19 |
Other References:
GONZALEZ J E ET AL: "IMPROVED INDICATORS OF CELL MEMBRANE POTENTIAL THAT USE FLUORESCENCE RESONANCE ENERGY TRANSFER", CHEMISTRY AND BIOLOGY,CURRENT BIOLOGY, LONDON,GB, vol. 4, no. 4, 1997, pages 269 - 277, XP000961796, ISSN: 1074-5521
CACCIATORE T W ET AL: "IDENTIFICATION OF NEURAL CIRCUITS BY IMAGING COHERENT ELECTRICAL ACTIVITY WITH FRET-BASED DYES", NEURON,CAMBRIDGE, MA,US, vol. 23, no. 3, July 1999 (1999-07-01), pages 449 - 459, XP000961663
GONZALEZ J E ET AL: "VOLTAGE SENSING BY FLUORESCENCE RESONANCE ENERGY TRANSFER IN SINGLECELLS", BIOPHYSICAL JOURNAL,NEW YORK, US,US, vol. 69, no. 4, 1 October 1995 (1995-10-01), pages 1272 - 1280, XP000608002, ISSN: 0006-3495
CACCIATORE T W ET AL: "IDENTIFICATION OF NEURAL CIRCUITS BY IMAGING COHERENT ELECTRICAL ACTIVITY WITH FRET-BASED DYES", NEURON,CAMBRIDGE, MA,US, vol. 23, no. 3, July 1999 (1999-07-01), pages 449 - 459, XP000961663
GONZALEZ J E ET AL: "VOLTAGE SENSING BY FLUORESCENCE RESONANCE ENERGY TRANSFER IN SINGLECELLS", BIOPHYSICAL JOURNAL,NEW YORK, US,US, vol. 69, no. 4, 1 October 1995 (1995-10-01), pages 1272 - 1280, XP000608002, ISSN: 0006-3495
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