Screening assay to identify modulators of the sleep/wake cycle

Abstract

Screening assays for identifying agents that modulate BK channel activity and further modulate the sleep/wake cycle in a subject, circadian regulated locomotor activity in a subject, or both are provided, as are agents identified using such screening assays. Also provided are methods of modulating the sleep/wake cycle in a subject and methods of modulating circadian regulated locomotor activity in a subject by administering an agent that modulates BK channel activity to the subject, for example, an agent identified by a screening assay as disclosed.

1 - 38 . (canceled) 
     
     
         39 . A method of identifying an agent that can modulate circadian regulated locomotor activity in a subject, the method comprising:
 contacting a test system comprising a calcium dependent potassium channel (BK channel) with an agent suspected of having the ability to modulate circadian regulated locomotor activity in the subject;   detecting a change in activity of the BK channel in the presence of the agent as compared to the activity of the BK channel in the absence of the agent, thereby identifying an agent that modulates BK channel activity;   administering the agent that modulates BK channel activity to a test subject; and   thereafter detecting a change in circadian regulated locomotor activity of the test subject, thereby identifying an agent that can modulate circadian regulated locomotor activity in a subject.   
     
     
         40 . The method of  claim 39 , wherein the BK channel comprises a  Drosophila  slowpoke (slo) polypeptide or an ortholog thereof. 
     
     
         41 . The method of  claim 40 , wherein the ortholog is a mammalian ortholog of the slo polypeptide. 
     
     
         42 . The method of  claim 39 , wherein the BK channel comprises a mutant BK channel. 
     
     
         43 . The method of  claim 39 , wherein the test system comprises a substantially purified BK channel polypeptide, and wherein said contacting is performed in vitro. 
     
     
         44 . The method of  claim 39 , wherein the test system comprises a membrane containing the BK channel, and wherein said contacting is performed in vitro. 
     
     
         45 . The method of  claim 44 , wherein the membrane is a muscle cell membrane or a nerve cell membrane. 
     
     
         46 . The method of  claim 39 , wherein the test system comprises a cell delimited by a cell membrane, or a cell membrane isolated from said cell, and wherein the BK channel is expressed in the cell membrane. 
     
     
         47 . The method of  claim 46 , wherein the BK channel is endogenous to the cell. 
     
     
         48 . The method of  claim 46 , wherein the BK channel is expressed from a heterologous nucleic acid molecule. 
     
     
         49 . The method of  claim 46 , wherein the cell is a muscle cell or a nerve cell. 
     
     
         50 . The method of  claim 46 , wherein the cell is a human cell. 
     
     
         51 . The method of  claim 48 , wherein the BK channel is a  Drosophila  slo polypeptide or an ortholog thereof. 
     
     
         52 . The method of  claim 46 , wherein the cell further expresses a BK channel binding protein. 
     
     
         53 . The method of  claim 52 , wherein the BK channel binding protein is a  Drosophila  slob polypeptide or an ortholog thereof. 
     
     
         54 . The method of  claim 52 , wherein the BK channel binding protein is a ∂ subunit of a BK channel. 
     
     
         55 . The method of  claim 39 , wherein the agent is a polynucleotide, a peptide, a peptidomimetic, a peptoid, or a small organic molecule. 
     
     
         56 . An agent identified by the method of  claim 39 . 
     
     
         57 - 67 . (canceled)