Compositions and Methods For Modulating Dopamine Nerutransmission

Abstract

The present invention provides for diagnosis or treatment of neurological or neuropsyciatric disorders involving abnormal dopamine neurotransmission. Methods and agents are provided for modulating dopamine transporter activity and modulating dopaminergic neurotransmission. Agents of the present invention include fragments of D2 receptor or dopamine transporter (DAT) that can disrupt D2-DAT coupling.

1 - 34 . (canceled) 
     
     
         35 . A method for increasing dopaminergic neurotransmission in a mammal in need of such treatment comprising administering a therapeutically effective amount of an agent that disrupts D2-DAT coupling in a mammal. 
     
     
         36 . The method of  claim 35 , wherein the agent is an antibody that binds to an amino acid sequence that is at least 80% identical to the sequence of D2[IL3-2-5] (SEQ ID NO: 1) or DAT[NT1-I] (SEQ ID NO:2). 
     
     
         37 . The method of  claim 36 , wherein the amino acid sequence is identical to the sequence of D2[IL3-2-5] (SEQ ID NO: 1) or the sequence DAT[N1-1] (SEQ ID NO: 1), wherein the antibody is optionally fused to a protein transduction domain. 
     
     
         38 . The method of  claim 35 , wherein the agent is a nucleic acid encoding a polypeptide of between about 7 and about 110 amino acids comprising an amino acid sequence that is at least 80% identical to the sequence of D2[IL3-2-5] (SEQ ID NO:1) or the sequence DAT[N1-I] (SEQ ID NO:2). 
     
     
         39 . The method of  claim 38 , wherein the polypeptide comprises an amino acid sequence that is identical to a sequence selected from the group consisting of D2[IL3-2] (SEQ ID NO:3), D2[IL3-2-3] (SEQ ID NO:4), D2[IL3-2-5] (SEQ ID NO:1), DAT[NT] (SEQ ID NO:6), DAT[NT1] (SEQ ID NO:7), and DAT[N1-I] (SEQ ID NO:2), wherein the antibody is optionally fused to a protein transduction domain. 
     
     
         40 . The method of  claim 39 , wherein the nucleic acid further encodes a protein transduction domain and the protein transduction domain is fused to the polypeptide. 
     
     
         41 . The method of  claim 35 , wherein the agent is a polypeptide of between about 7 and about 10 amino acids comprising an amino acid sequence that is between about 80% and about 100% identical to the sequence of D2[IL3-2-5] (SEQ ID NO: 1) or the sequence of DAT[N1-1] (SEQ ID NO:2), and the polypeptide is optionally fused to a protein transduction domain. 
     
     
         42 . The method of  claim 41 , wherein the polypeptide comprises an amino acid sequence that is identical to a sequence selected from the group consisting of D2[IL3-2] (SEQ ID NO:3), D2[IL3-2-3] (SEQ ID NO:4), D2[IL3-2-5] (SEQ ID NO: 1), DAT[NT] (SEQ ID NO:6), DAT[NT1] (SEQ ID NO:7), and DAT[N1-I] (SEQ ID NO:1). 
     
     
         43 . The method of  claim 35 , wherein the agent is administered for treating a disease selected from the group consisting of psychostimulant addiction including cocaine addiction or amphetamine addiction, depression, obesity, ADHD, narcolepsy and Parkinson's disease. 
     
     
         44 . A method for reducing dopaminergic neurotransmission in a mammal in need of such treatment comprising administering a therapeutically effective amount of an agent that localizes DAT at the cell surface. 
     
     
         45 . The method of  claim 44 , wherein the agent is a nucleic acid encoding a D2 receptor or a derivative thereof that is between about 70% and 100% identical to the D2 receptor and that maintains DAT-coupling activity, wherein the nucleic acid is optionally fused to a protein transduction domain. 
     
     
         46 . The method of  claim 45 , wherein the agent is a polypeptide comprising a D2 receptor or a derivative thereof that is between about 70% and 100% identical to the D2 receptor and that maintains DAT-coupling activity, wherein the polypeptide is optionally fused to a protein transduction domain. 
     
     
         47 . The method of  claim 44 , wherein the agent is administered for treating a disease selected from the group consisting of schizophrenia, and Tourette's syndrome. 
     
     
         48 . A method of diagnosing schizophrenia or depression in a patient comprising determining whether cell surface localization of DAT in a test sample from the patient is less than in control samples. 
     
     
         49 . The method of  claim 48 , wherein cell surface localization of DAT is determined by quantifying dopamine uptake. 
     
     
         50 . The method of  claim 48 , wherein cell surface localization of DAT is determined by immunolabeling with an antibody specific for DAT. 
     
     
         51 . The method of  claim 48 , wherein cell surface localization of DAT is determined by coimmunoprecipitation of DAT with an antibody specific for D2 receptor. 
     
     
         52 . The method of  claim 48 , wherein cell surface localization of DAT is determined by affinity precipitation of DAT with a D2 receptor. 
     
     
         53 . The method of  claim 48 , wherein cell surface localization of DAT is determined by quantifying DAT ligand binding. 
     
     
         54 . A polypeptide of between about 7 and about 110 amino acids comprising an amino acid sequence that is at between about 80% and about 100% identical to the sequence of D2[IL3-2-5] (SEQ ID NO: 1), DAT[N1-I] (SEQ ED NO:2) or DAT[NT1-2] (SEQ ID NO: 11), and optionally comprising a protein transduction domain, wherein the protein transduction domain is selected from the group consisting of TAT, and SynB1/3Cit. 
     
     
         55 . The polypeptide of  claim 54 , comprising an amino acid sequence that is between about 80% and 100% identical to a sequence selected from the group consisting of D2[IL3-2] (SEQ ID NO:3), D2[IL3-2-3] (SEQ ID NO:4), D2[IL3-2-5] (SEQ ID NO:1), DAT[NT] (SEQ ID NO:6), DAT[NT1] (SEQ ID NO:7), DAT[N1-I] (SEQ ID NO:2) and DAT[NT1-2] (SEQ ID NO:1). 
     
     
         56 . The polypeptide of  claim 54 , comprising an amino acid sequence that is identical to a sequence selected from the group consisting of D2[IL3-2] (SEQ ID NO:3), D2[IL3-2-3] (SEQ ID NO:4), D2[IL3-2-5] (SEQ ID NO:1), DAT[NT] (SEQ ID NO:6), DAT[NT1] (SEQ ID NO:7), DAT[N1-I] (SEQ ID NO:2) and DAT[NT1-2] (SEQ ID NO:11). 
     
     
         57 . A nucleic acid encoding a polypeptide of between 7 and 110 amino acids comprising an amino acid sequence that is between about 80% identical and 100% identical to the sequence of D2[IL3-2-5] (SEQ ID NO: 1) or the sequence of DAT[N1-I] (SEQ ID NO:2), wherein the nucleic acid optionally encodes a protein transduction domain and the protein transduction domain is optionally fused to the polypeptide, wherein the protein transduction domain, when present, is selected from the group consisting of TAT, and SynB1/3Cit. 
     
     
         58 . The nucleic acid of  claim 57 , wherein the polypeptide comprises an amino acid sequence that is between 80% and 100% identical to a sequence selected from the group consisting of D2[IL3-2] (SEQ ID NO:3), D2[IL3-2-3] (SEQ ID NO:4), D2[IL3-2-5] (SEQ ID NO:1), DAT[NT] (SEQ ID NO:6), DAT[NT1] (SEQ ID NO:7), DAT[N1-I] (SEQ ID NO:2) and DAT[NT1-2] (SEQ ID NO:11). 
     
     
         59 . The nucleic acid of  claim 57 , wherein the polypeptide comprises an amino acid sequence that is identical to a sequence selected from the group consisting of D2[IL3-2] (SEQ ID NO:3), D2[IL3-2-3] (SEQ ID NO:4), D2[IL3-2-5] (SEQ ID NO: 1), DAT[NT] (SEQ ID NO:6), DAT[NT1] (SEQ ID NO:7), DAT[N1-I] (SEQ ID NO:2) and DAT[NT1-2] (SEQ ID NO: 11).