Methods of screening agents for activity using teleosts

Abstract

The present invention provides methods of screening an agent for activity using teleosts. Methods of screening an agent for angiogenesis activity, toxic activity and an effect cell death activity in teleosts are provided. Methods of screening an agent for an activity in the brain or central nervous system in zebrafish are provided. The invention further provides high throughput methods of screening agents in multi-well plates.

1 . A method of screening an agent for an effect on a brain or central nervous system (CNS) disease or disorder in a zebrafish in vivo, the method comprising:
 (a) administering a compound to the zebrafish, wherein the compound induces a phenotype in the zebrafish that mimics the brain or CNS disease or disorder;   (b) administering the agent to the zebrafish;   (c) detecting a response in the brain or CNS of the zebrafish; and   (d) comparing the response in the brain or CNS of the agent-treated zebrafish with a response in the brain or CNS of the compound-treated zebrafish administered with no agent or a control agent, wherein a difference in the response in the brain or CNS of the agent-treated zebrafish compared to the response in the brain or CNS of the untreated or control agent-treated zebrafish indicates the agent has an effect on the brain or CNS disease or disorder.   
     
     
         2 . A method of screening an agent for an ability to affect blood brain barrier (BBB) permeability in a zebrafish in vivo, the method comprising:
 (a) administering the agent to the zebrafish of age at least 3 days post-fertilization;   (b) detecting a response in the zebrafish; and   (c) comparing the response in the agent-treated zebrafish with a response in a zebrafish administered with no agent or a control agent, wherein a difference in the response in the agent-treated zebrafish compared the untreated or control agent-treated zebrafish indicates the agent has the ability to affect BBB permeability.   
     
     
         3 .- 10 . (canceled) 
     
     
         11 . The method of  claim 2 , wherein the agent is administered to the zebrafish by dissolving the agent in the media containing the zebrafish. 
     
     
         12 . The method of  claim 2 , wherein the agent is injected into the zebrafish. 
     
     
         13 . The method of  claim 2 , wherein the agent is administered by direct delivery to the brain or CNS of the zebrafish. 
     
     
         14 - 18 . (canceled) 
     
     
         19 . The method of  claim 2 , wherein the agent is a small molecule and a library of small molecules is screened for permeability of the BBB. 
     
     
         20 . The method of  claim 2 , wherein the agent is a nucleic acid, peptide, protein, glycoprotein, carbohydrate, lipid, or glycolipid. 
     
     
         21 . The method of  claim 20 , wherein the nucleic acid is DNA or RNA. 
     
     
         22 - 24 . (canceled) 
     
     
         25 . The method of  claim 2 , wherein the zebrafish is an embryo, larva, or adult. 
     
     
         26 . The method of  claim 25 , wherein the zebrafish is a zebrafish embryo. 
     
     
         27 . The method of  claim 2 , wherein the zebrafish is a wild-type strain. 
     
     
         28 . The method of  claim 2 , wherein the zebrafish is a transgenic zebrafish. 
     
     
         29 . The method of  claim 2 , wherein the zebrafish has a heritable mutation. 
     
     
         30 . The method of  claim 29 , wherein the heritable mutation is associated with a brain or CNS disease or disorder. 
     
     
         31 . A method of screening an agent for a first activity in the brain or central nervous system (CNS), and for a second activity in at least one other tissue or organ, of a zebrafish in vivo, the method comprising:
 (a) administering the agent to the zebrafish;   (b) detecting a first response in the brain or CNS;   (c) comparing the first response in the brain or CNS of the agent-treated zebrafish with the first response in the brain or CNS of a zebrafish administered with no agent or a control agent,   wherein a difference in the first response in the brain or CNS of the agent-treated zebrafish compared to the first response in the brain or CNS of the untreated or control agent-treated zebrafish indicates the agent has the first activity in the brain or CNS;   (d) detecting a second response in the at least one other tissue or organ of the zebrafish; and   (e) comparing the second response in the at least one other tissue or organ of the agent-treated zebrafish with the second response in the at least one other tissue or organ of the zebrafish administered with no agent a control agent,   wherein a difference in the second response in the at least one other tissue or organ of the agent-treated zebrafish compared to the second response in the at least one other tissue or organ of the untreated or control agent-treated zebrafish indicates the agent has the second activity in the at least one other tissue or organ.   
     
     
         32 . The method of  claim 31 , wherein the first activity and the second activity are the same. 
     
     
         33 . The method of  claim 31 , wherein the first activity and the second activity are different. 
     
     
         34 . A method of screening an agent for an ability to affect blood brain barrier (BBB) permeability, and for an activity in at least one other tissue or organ, of a zebrafish in vivo, the method comprising
 (a) administering an agent in vivo to a zebrafish;   (b) detecting a first response in the zebrafish;   (c) comparing the first response in the agent-treated zebrafish with the first response in a zebrafish administered with no agent or a control agent, wherein a difference in the first response in the agent-treated zebrafish compared to the first response in the untreated or control agent-treated zebrafish indicates the agent has the ability to affect BBB permeability;   (d) detecting a second response in the at least one other tissue or organ; and   (e) comparing the second response in the at least one other tissue or organ of the agent-treated zebrafish with the second response in the at least one other tissue or organ of a zebrafish administered with no agent or a control agent, wherein a difference in the second response in the agent-treated zebrafish compared to the second response in the at least one other tissue or organ of the untreated or control agent-treated zebrafish indicates the agent has the activity in the at least one other tissue or organ.   
     
     
         35 . The method of  claim 34 , wherein the at least one other tissue or organ is the brain or CNS. 
     
     
         36 . The method of  claim 34 , wherein the at least one other tissue or organ is other than the brain or CNS.