Methods of screening agents for activity using teleosts

• US 7,951,989 B2
• Filed: 09/01/2006
• Issued: 05/31/2011
• Est. Priority Date: 02/23/1998
• Status: Expired due to Fees

• Current Assignee: Phylonix Pharmaceuticals Incorporated
• Original Assignee: Phylonix Pharmaceuticals Incorporated
• Inventors: Parng, Chuenlei, Serbedzija, George N., McGrath, Patricia
• Primary Examiner(s): BERTOGLIO, VALARIE E
• Application Number:
  US11/515,355
• Publication Number:
  US 20070143865A1
• Time in Patent Office: 1,733 Days
• Field of Search: 800/20
• US Class Current: 800/20
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/40 Fish
• CPC Class Codes: A01K 2267/0393 Animal model comprising a reporter system for screening tests
• CPC Class Codes: A61K 49/0008 Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
• CPC Class Codes: A61K 49/006 Biological staining of tissues in vivo, e.g. methylene blue or toluidine blue O administered in the buccal area to detect epithelial cancer cells, dyes used for delineating tissues during surgery
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: G01N 2333/4603 from fish
• CPC Class Codes: G01N 2510/00 Detection of programmed cell death, i.e. apoptosis
• CPC Class Codes: G01N 33/5014 for testing toxicity
• CPC Class Codes: G01N 33/5088 of vertebrates

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.

Claims

1. A method of screening an agent in vivo for a toxic activity in the brain or central nervous system (CNS), the method comprising:

(a) administering the agent to a teleost;

(b) detecting cell death, if any, in the brain or CNS of the teleost; and

(c) comparing the cell death, if any, in the brain or CNS of the agent-treated teleost with cell death, if any in the brain or CNS of an untreated or control teleost, wherein an increase in cell death in the agent-treated teleost compared to the untreated or control teleost identifies the agent as having toxic activity in the brain or CNS.

2. The method of claim 1, wherein the agent is administered to the teleost by dissolving the agent in the media containing the teleost.

3. The method of claim 1, wherein the agent is injected into the teleost.

4. The method of claim 1, wherein the agent is administered by direct delivery to the brain or CNS of the teleost.

5. The method of claim 4, wherein the direct delivery to the brain or CNS is microinjection of the agent into the brain or CNS of the teleost.

6. The method of claim 1, wherein the agent is administered to the teleost in conjunction with a carrier.

7. The method of claim 6, wherein the carrier is a solvent, lipid, or peptide.

8. The method of claim 1, wherein the agent is administered to the teleost in conjunction with at least one other compound.

9. The method of claim 1, wherein the agent is fluorescent or radioactive.

10. The method of claim 1, wherein the agent is a small molecule and a library of small molecules is screened for toxic activity in the brain or CNS.

11. The method of claim 1, wherein the agent is a nucleic acid, peptide, protein, glycoprotein, carbohydrate, lipid, or glycolipid.

12. The method of claim 1, wherein the teleost is an embryo, larva, or adult.

13. The method of claim 12, wherein the teleost is an embryo.

14. The method of claim 1, wherein the teleost is a wild-type strain.

15. The method of claim 1, wherein the teleost is transgenic.

16. The method of claim 1, wherein the teleost has a heritable mutation.

17. The method of claim 16, wherein the heritable mutation is associated with a brain or CNS disease or disorder.

18. The method of claim 1, wherein the teleost is a zebrafish.

19. The method of claim 1, wherein the agent is administered to the teleost within 24 hours post fertilization.

20. The method of claim 1, wherein the agent is administered to the teleost within 48 hours post fertilization.

21. The method of claim 1, wherein the agent is administered to the teleost within 72 hours post fertilization.

22. The method of claim 1, wherein the agent is a nucleic acid analogue.

23. The method of claim 1, wherein the cell death is apoptosis.