Claims
- 1. A method for identifying a compound that increases or decreases neuritic growth, said method comprising the steps of:
(a) contacting a culture comprising neurons with a test compound; and (b) measuring the amount of a protein or RNA that is preferentially found in the neurites of said neurons, wherein an increase in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that increases neuritic growth and a decrease in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that decreases neuritic growth.
- 2. The method of claim 1, wherein said protein is selected from the group consisting of alpha-tubulin, beta-tubulin, GAP-43, neurofilament light chain, neurofilament medium chain, neurofilament heavy chain, microtubule-associated protein (MAP) 1B, MAP4, MAP2C, and MAP2D.
- 3. The method of claim 1, wherein said neurons comprise sympathetic neurons, cholinergic neurons, sensory neurons, cortical neurons, motor neurons, and/or dorsal root ganglion neurons.
- 4. The method of claim 1, wherein said neurons comprise primary neurons, neurons derived from a cell line, and/or neurons derived from a stem cell.
- 5. The method of claim 1, wherein the test compound is selected from the group consisting of a peptide or protein, a carbohydrate, a lipid, a nucleic acid molecule, a natural organic molecule, and a synthetically derived organic molecule.
- 6. A method for identifying a compound that increases or decreases dendritic growth, said method comprising the steps of:
(a) contacting a culture comprising neurons with a test compound; and (b) measuring the amount of a protein or RNA that is preferentially found in the dendrites of said neurons, wherein an increase in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that increases dendritic growth and a decrease in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that decreases dendritic growth.
- 7. The method of claim 6, wherein said protein is selected from the group consisting of MAP2A, MAP2B, dendrin, type II calcium-calmodulin-dependent protein (CamIIK), metabotropic glutamate receptor (mGluR) 1A, mGluR2, and type 1A serotonin receptor.
- 8. The method of claim 6, wherein said RNA is selected from the group consisting of MAP2A RNA, MAP2B RNA, dendrin RNA, and CamIIK RNA.
- 9. A method for identifying a compound that increases or decreases axonal growth, said method comprising the steps of:
(a) contacting a culture comprising neurons with a test compound; and (b) measuring the amount of a protein or RNA that is preferentially found in the axons of said neurons, wherein an increase in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that increases axonal growth and a decrease in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that decreases axonal growth.
- 10. The method of claim 9, wherein said protein is selected from the group consisting of phosphorylated MAP1B, phosphorylated neurofilament heavy chain, and type 1B serotonin receptor.
- 11. A method of identifying a compound that increases or decreases neuronal growth, said method comprising the steps of:
(a) contacting a culture comprising neurons with NGF at levels sufficient to support cell survival; (b) contacting said culture with a neuronal growth-inhibiting compound; (c) contacting said culture with a test compound; and (d) measuring the amount of a protein or RNA that is preferentially found in the axons, dendrites, or neurites of said neurons, wherein an increase in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that increases neuronal growth and a decrease in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that decreases neuronal growth.
- 12. The method of claim 11, wherein said neuronal growth-inhibiting compound is BDNF or an inhibitor of MEK signaling.
- 13. The method of claim 12, wherein said inhibitor of MEK signaling is a dominant-negative MEK.
- 14. The method of claim 12, wherein said inhibitor of MEK signaling is U0126 or PD98059.
- 15. A method of identifying a compound that increases or decreases neuronal growth, said method comprising the steps of:
(a) inhibiting MEK signaling in a culture comprising neurons; (b) contacting said culture with a test compound; and (c) measuring the amount of a protein or RNA that is preferentially found in the axons, dendrites, or neurites of said neurons, wherein an increase in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that increases neuronal growth and a decrease in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that decreases neuronal growth.
- 16. The method of claim 15, wherein said MEK signaling is inhibited by contacting said neuron with an inhibitor of MEK signaling.
- 17. The method of claim 16, wherein said inhibitor of MEK signaling is a dominant-negative MEK.
- 18. The method of claim 16, wherein said inhibitor of MEK signaling is U0126 or PD98059.
- 19. A method of identifying a compound that increases neuronal growth, said method comprising:
(a) activating p75NTR signaling in a culture comprising neurons; (b) contacting said culture with a test compound; and (c) measuring the amount of a protein or RNA that is preferentially found in the axons, dendrites, or neurites of said neurons, wherein an increase in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that increases neuronal growth and a decrease in the amount of said protein or RNA relative to the amount of said protein or RNA in said culture not contacted with said test compound identifies said test compound as a compound that decreases neuronal growth.
- 20. The method of claim 19, wherein said p75NTR signaling is activated by introducing ceramide into said neuron.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims benefit from U.S. Provisional Application Serial No. 60/203,560, filed May 11, 2000 (now pending), which is hereby incorporated by reference.
Provisional Applications (1)
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Number |
Date |
Country |
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60203560 |
May 2000 |
US |