Methods of modulating cold sensory perception via a cold- and menthol-sensitive receptor (CMR1)
The present invention relates to regulation of cold sensation and pain. More particularly, the present invention is directed to nucleic acids encoding a member of the transient regulatory protein family, CMR1, which is involved in modulation of the perception of cold sensations and pain. The invention further relates to methods for identifying and using agents that modulate cold responses and pain responses stimulated by cold via modulation of CMR1 and CMR1-related signal transduction.
1. A method for identifying a compound that modulates a cool or cold sensation in a mammalian subject, the method comprising:
(i) contacting a cell comprising a cold- and menthol-sensitive receptor (CMR1) polypeptide with the compound, wherein the CMR1polypeptide comprises a sequence having at least 90% identity to SEQ ID NO:3 and forms a cation channel; and
(ii) determining whether said compound that modulates the activity of said CMR1polypeptide based on an effect in said cell that correlates to CMR1activity, and
(iii) if said compound elicits the effect in said cell, further determining whether said compound modulates a cool/cold sensation in a mammalian subject, thereby identifying a compound that modulates cool or cold-sensation.
2. The method of claim 1 , wherein the effect is a change in intracellular calcium.
3. The method of claim 2 , wherein the effect is measured using a fluorescent compound.
4. The method of claim 1 , wherein the effect is a change in membrane potential.
5. The method of claim 1 , wherein the cell is a non-neuronal cell.
6. The method of claim 1 , wherein the CMR1is recombinant.
7. The method of claim 1 , wherein the CMR1polypeptide is encoded by a nucleic acid having at least 95% identity to SEQ ID NO: 4.
8. The method of claim 1 , wherein the CMR1polypeptide is encoded by a nucleic acid having at least 98% identity to SEQ ID NO: 4.
9. The method of claim 1 , wherein the CMR1polypeptide is encoded by a nucleic acid having the sequence set forth in SEQ ID NO: 4.
10. The method of claim 1 , further comprising a step of stimulating the cell with cold temperature.
11. The method of claim 1 , wherein the identified compound enhances cool or cold sensation.
12. The method of claim 1 , wherein the identified compound inhibits cool or cold sensation.
13. The method of claim 1 , wherein the cell is derived from mammalian cell lines 293 or CHO.
14. The method of claim 1 , wherein the CMR1polypeptide has at least 95% identity to SEQ ID NO: 3.
15. The method of claim 1 , wherein the CMR1polypeptide has at least 98% identity to SEQ ID NO: 3.
16. The method of claim 1 , wherein the CMR1polypeptide has the amino acid sequence set forth in SEQ ID NO: 3.
17. The method of claim 1 , wherein said mammal is a human.
18. The method of claim 1 , wherein the cation channel responds to cool or cold temperatures.
19. The method of claim 1 , wherein the cation channel responds to a menthol compound.