METHODS AND KITS FOR IDENTIFICATION OF ANTI-EXCITOTOXIC COMPOUNDS
First Claim
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1. A method of identifying compounds reducing excitotoxicity in neurons, the method comprising:
- a) providing a library of compounds suspected of reducing excitotoxicity in neurons;
b) providing a plurality of samples of glial cells;
c) contacting at least one member of the plurality of the samples with at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons;
d) determining the amount of EAAT-1 protein in said at least one member of the plurality of the samples of glial cells contacted with the at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons;
e) comparing the amount of the EAAT-1 protein from step (d) with the amount of EAAT protein from at least one member of the plurality of samples of glial cells not contacted with the at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons; and
f) selecting the members of the library of compounds suspected of reducing excitotoxicity in neurons which increase the amount of the EAAT protein in the respective members of the plurality of samples of glial cells.
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Abstract
The instant invention provides methods and kits for screening for anti-excitotoxic compounds which increase the total amount of EAAT-I protein or the surface amount of EAAT-I protein.
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Citations
20 Claims
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1. A method of identifying compounds reducing excitotoxicity in neurons, the method comprising:
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a) providing a library of compounds suspected of reducing excitotoxicity in neurons; b) providing a plurality of samples of glial cells; c) contacting at least one member of the plurality of the samples with at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons; d) determining the amount of EAAT-1 protein in said at least one member of the plurality of the samples of glial cells contacted with the at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons; e) comparing the amount of the EAAT-1 protein from step (d) with the amount of EAAT protein from at least one member of the plurality of samples of glial cells not contacted with the at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons; and f) selecting the members of the library of compounds suspected of reducing excitotoxicity in neurons which increase the amount of the EAAT protein in the respective members of the plurality of samples of glial cells. - View Dependent Claims (2, 5, 6, 7, 13, 17)
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3. A method of identifying compounds reducing excitotoxicity in neurons, the method comprising:
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a) providing a library of compounds suspected of reducing excitotoxicity in neurons; b) providing a plurality of samples of glial cells; c) contacting at least one member of the plurality of the samples with at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons; d) determining the amount of EAAT-1 protein present on a cell surface of said at least one member of the plurality of the samples of glial cells contacted with the at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons; e) comparing the amount of the EAAT-1 protein from step (d) with the amount of EAAT protein present on a cell surface of at least one member of the plurality of samples of glial cells not contacted with the at least one member of the library of the compounds suspected of reducing excitotoxicity in neurons; and f) selecting the members of the library of compounds suspected of reducing excitotoxicity in neurons which increase the amount of the EAAT protein on the respective cell surfaces of the respective members of the plurality of samples of glial cells. - View Dependent Claims (4, 14, 15)
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8. A kit for of identifying compounds reducing excitotoxicity in neurons, the method comprising:
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a) a plurality of samples of glial cells; b) a means for determining the amount of EAAT-1 protein present on a cell surface; and c) a set of instructions. - View Dependent Claims (10, 11, 12, 16, 19, 20)
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9. A kit for of identifying compounds reducing excitotoxicity in neurons, the method comprising:
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a) a plurality of samples of glial cells; b) a means for determining the amount of EAAT-1 protein present in a cell; and c) a set of instructions. - View Dependent Claims (18)
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Specification