Patent ID: 6808873
Filing Date: 2004-10-26
Classification: G01N

Abstract:
A method of identifying an agent that alters mitochondrial function, comprising:(a) contacting, in each of a plurality of reaction vessels in a high throughput screening array, (i) a biological sample comprising a cell containing cytosol, a mitochondrion and a calcium indicator molecule, under conditions that permit maintenance of mitochondrial membrane potential, with (ii) a source of calcium cations, wherein the calcium indicator molecule is capable of generating a detectable signal that is proportional to the level of calcium in the cytosol; (b) detecting in each reaction vessel the signal generated by the calcium indicator molecule at a plurality of time points, under conditions that permit identification of (i) mitochondrial calcium uniporter activity, wherein a decrease over time in the signal that is proportional to the level of calcium in the cytosol indicates mitochondrial calcium uniporter activity, and (ii) mitochondrial uncoupler or respiratory inhibitor activity, wherein an increase over time in the signal that is proportional to the level of calcium in the cytosol without repeating step (a) indicates mitochondrial uncoupler or respiratory activity; and (c) comparing the signal generated by the calcium indicator molecule at one or more of said time points in the absence of a candidate agent, to the signal generated by the calcium indicator molecule at one or more of said time points in the presence of the candidate agent, and therefrom identifying an agent that alters mitochondrial function.