Use of the conserved Drosophila NPFR1 system for uncovering interacting genes and pathways important in nociception and stress response
First Claim
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1. A method of identifying a composition capable of inhibiting a response to a stressor, comprising:
- exposing a Drosophila melanogaster organism to a medium comprising a compound that elicits an avoidance response in a wild-type Drosophila organism, wherein the Drosophila organism exhibits an avoidance response to the medium; and
contacting the Drosophila organism with a test compound, wherein a reduction in the avoidance response to the medium in the presence of the test compound as compared to in the absence of the test compound indicates that the test compound modulates response to a stressor in a higher organism.
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Abstract
The present disclosure provides methods of identifying compositions capable of inhibiting responses to stressors in an organism, as well as recombinant cells and organisms useful in identifying compositions capable of inhibiting responses to stressors in an organism.
42 Citations
46 Claims
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1. A method of identifying a composition capable of inhibiting a response to a stressor, comprising:
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exposing a Drosophila melanogaster organism to a medium comprising a compound that elicits an avoidance response in a wild-type Drosophila organism, wherein the Drosophila organism exhibits an avoidance response to the medium; and contacting the Drosophila organism with a test compound, wherein a reduction in the avoidance response to the medium in the presence of the test compound as compared to in the absence of the test compound indicates that the test compound modulates response to a stressor in a higher organism. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A method of identifying a composition capable of modulating a response to a stressor, comprising:
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providing a recombinant Drosophila melanogaster organism comprising a heterologous transient receptor potential (TRP) ion-channel polypeptide from a different organism, wherein the TRP ion-channel polypeptide is responsive to a stressor; exposing the larva to the stressor, wherein the Drosophila larva exhibits an avoidance response to the stressor; and contacting the larva with a test compound, wherein a reduction in the avoidance response to the stressor in the presence of the test compound as compared to in the absence of the test compound indicates that the test compound modulates response to a stressor in a higher organism. - View Dependent Claims (9, 10)
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11. A recombinant Drosophila melanogaster organism comprising:
- a heterologous transient receptor potential (TRP) ion-channel polypeptide from a different organism, wherein the TRP ion-channel polypeptide is responsive to a stressor.
- View Dependent Claims (12)
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13. A method of identifying a composition capable of inhibiting a response to a stressor, comprising:
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providing a recombinant Drosophila melanogaster organism comprising neurons comprising a nucleic acid encoding for a transient receptor potential (TRP) ion-channel polypeptide that is responsive to a stressor, wherein the neurons further comprise a heterologous nucleic acid encoding a neuropeptide family receptor; exposing the organism to the stressor and observing the organism'"'"'s response to the stressor; exposing the organism to the stressor in the presence of a test compound; and observing the organism'"'"'s response to the stressor, wherein a change in the organism'"'"'s response to the stressor in the presence of the test compound as compared to the response in the absence of the test compound indicates that the test compound modulates the response to the stressor. - View Dependent Claims (14, 15, 16)
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17. A recombinant Drosophila melanogaster organism comprising a neural cell comprising:
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a nucleic acid encoding for a transient receptor potential (TRP) ion-channel polypeptide that is responsive to a particular stressor, and a heterologous nucleic acid encoding a neuropeptide family receptor that is not present in the neural cell of a wild-type Drosophila organism.
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18. A method of identifying a composition capable of reducing a cellular response to a stressor, comprising:
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exposing a first population of cells to a stressor, wherein the cells comprise a heterologous nucleic acid encoding a transient receptor potential (TRP) ion-channel polypeptide and a heterologous nucleic acid encoding a neuropeptide receptor, and a fluorescent compound capable of intracellularly fluorescing in the presence of calcium, wherein the TRP ion-channel polypeptide transports calcium into the cell in response to a stressor; delivering to a second population of cells a stressor, wherein the cells comprise the heterologous nucleic acid encoding the TRP ion-channel polypeptide, and the heterologous nucleic acid encoding the neuropeptide receptor, the fluorescent compound is capable of intracellularly fluorescing in the presence of calcium, and a test compound; and determining the difference in the level fluorescence of the first and second cell populations, wherein if the level of fluorescence in the first cell population is greater than in the second cell population, the test compound inhibits the uptake of calcium via the transient receptor potential ion-channel polypeptide. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27)
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28. A method of identifying a composition capable of reducing a cellular response to a stressor, comprising:
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exposing a first population of cells to a stressor, wherein the cells comprise a heterologous nucleic acid encoding a transient receptor potential (TRP) ion-channel polypeptide and a heterologous nucleic acid encoding a neuropeptide receptor, and wherein the cells are in contact with a medium comprising a neuropeptide capable of selectively binding to the neuropeptide receptor, and a fluorescent compound capable of intracellularly fluorescing in the presence of calcium, wherein the TRP ion-channel polypeptide transports calcium into the cell in response to a stressor; delivering to a second population of cells a stressor, wherein the cells comprise the heterologous nucleic acid encoding the TRP ion-channel polypeptide, and the heterologous nucleic acid encoding the neuropeptide receptor, and wherein the cells are in contact with a medium comprising the neuropeptide capable of selectively binding to the neuropeptide receptor, the fluorescent compound is capable of intracellularly fluorescing in the presence of calcium, and a test compound; and determining the difference in the level fluorescence of the first and second cell populations, wherein if the level of fluorescence in the first cell population is greater than in the second cell population, the test compound inhibits the uptake of calcium via the transient receptor potential ion-channel polypeptide. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37)
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38. A recombinant eukaryotic cell comprising:
- a heterologous nucleic acid encoding a transient receptor potential (TRP) ion-channel polypeptide, and a heterologous nucleic acid encoding a neuropeptide Y family receptor.
- View Dependent Claims (39, 40, 41, 42, 43, 44, 45, 46)
Specification