Hypothalamus-specific polypeptides

Abstract

Peptides and polypeptides found in the hypothalamus region of the mammalian brain are described, particularly hypocretin polypeptides and their uses. Hypocretin polypeptides are biologically active and produce electrical changes in neurons, lower body temperature, and reduce food intake.

We claim:  
     
         1 . An isolated polynucleotide selected from the group consisting of: 
 (a) a polynucleotide encoding the polypeptide comprising the amino acid sequence of SEQ ID NO: 1;    (b) a polynucleotide encoding the polypeptide comprising the amino acid sequence of SEQ ID NO: 2;    (c) a polynucleotide capable of hybridizing to and which is at least 95% homologous to the polynucleotide of (a) or (b).    
     
     
         2 . The polynucleotide of  claim 1  comprising the sequence of the group consisting of the second line of FIG. 5 and SEQ ID NO: 3.  
     
     
         3 . The polynucleotide of  claim 1  comprising the sequence of the group consisting of the third line of FIG. 5 and SEQ ID NO: 4.  
     
     
         4 . An isolated polynucleotide selected from the group consisting of: 
 (a) a polynucleotide encoding a polypeptide comprising the sequence of SEQ ID NO: 6;    (b) a polynucleotide encoding a polypeptide comprising amino acids 28 to 130 of SEQ ID NO: 2;    (c) a polynucleotide capable of hybridizing to the polynucleotide of (a);    (d) a polynucleotide capable of hybridizing to the polynucleotide of (b);    (e) a polynucleotide that is at least 95% homologous to the polynucleotide of (a); and    (f) a polynucleotide that is at least 95% homologous to the polynucleotide of (b).    
     
     
         5 . An isolated polynucleotide selected from the group consisting of: 
 (a) a polynucleotide encoding a polypeptide comprising the sequence of SEQ ID NO: 7;    (b) a polynucleotide encoding a polypeptide comprising the sequence of SEQ ID NO: 8;    (c) a polynucleotide encoding a polypeptide comprising amino acids 42 to 66 of SEQ ID NO: 1;    (d) a polynucleotide encoding a polypeptide comprising amino acids 42 to 65 of SEQ ID NO: 1;    (e) a polynucleotide encoding a polypeptide comprising amino acids 43 to 66 of SEQ ID NO: 1;    (f) a polynucleotide encoding a polypeptide comprising amino acids 43 to 65 of SEQ ID NO: 1; and    (g) a polynucleotide capable of hybridizing to and which is at least 95% homologous to a polynucleotide of (a) through (f).    
     
     
         6 . An isolated polynucleotide selected from the group consisting of: 
 (a) a polynucleotide encoding a polypeptide comprising the sequence of SEQ ID NO: 9;    (b) a polynucleotide encoding a polypeptide comprising the sequence of SEQ ID NO: 10; and    (c) a polynucleotide capable of hybridizing to and which is at least 95% homologous to the polynucleotide of (a) or (b).    
     
     
         7 . An isolated polynucleotide selected from the group consisting of: 
 (a) a polynucleotide encoding a polypeptide comprising amino acids 100 to 130 of SEQ ID NO: 1;    (b) a polynucleotide encoding a polypeptide comprising amino acids 100 to 130 of SEQ ID NO: 2;    (c) a polynucleotide encoding a polypeptide comprising amino acids 100 to 111 of SEQ ID NO: 1;    (d) a polynucleotide encoding a polypeptide comprising amino acids 100 to 110 of SEQ ID NO: 1;    (e) a polynucleotide encoding a polypeptide comprising amino acids 114 to 130 of SEQ ID NO: 1; and    (f) a polynucleotide capable of hybridizing to and which is at least 95% homologous to a polynucleotide of (a) through (e).    
     
     
         8 . An isolated polypeptide selected from the group consisting of the amino acid sequence of SEQ ID NO: 1 and the amide thereof.  
     
     
         9 . An isolated polypeptide selected from the group consisting of the amino acid sequence of SEQ ID NO: 2 and the amide thereof.  
     
     
         10 . An isolated polypeptide selected from the group consisting of a polypeptide comprising the sequence of SEQ ID NO: 6, a polypeptide comprising amino acids 28 to 130 of SEQ ID NO: 2, and the amides thereof.  
     
     
         11 . An isolated polypeptide comprising the sequence of SEQ ID NO: 13.  
     
     
         12 . The isolated peptide of  claim 11  wherein the isolated peptide is from about 28 amino acid residues to about 39 amino acid residues long.  
     
     
         13 . An isolated polypeptide selected from the group consisting of: 
 (a) a polypeptide comprising the sequence of SEQ ID NO: 7;    (b) a polypeptide comprising the sequence of SEQ ID NO: 8;    (c) a polypeptide comprising amino acids 42 to 66 of SEQ ID NO: 1;    (d) a polypeptide comprising amino acids 42 to 65 of SEQ ID NO: 1;    (e) a polypeptide comprising amino acids 43 to 66 of SEQ ID NO: 1;    (f) a polypeptide comprising amino acids 43 to 65 of SEQ ID NO: 1;    (g) a polypeptide comprising at least one conservative amino acid substitution in the sequence of polypeptides (a-f); and    (h) the amides thereof.    
     
     
         14 . An isolated polypeptide selected from the group consisting of: 
 (a) a polypeptide comprising the sequence of SEQ ID NO: 9;    (b) a polypeptide comprising the sequence of SEQ ID NO: 10; and    (c) the amides thereof.    
     
     
         15 . An isolated polypeptide selected from the group consisting of: 
 (a) a polypeptide comprising amino acids 100 to 130 of SEQ ID NO: 1;    (b) a polypeptide comprising amino acids 100 to 130 of SEQ ID NO: 2;    (c) a polypeptide comprising amino acids 100 to 111 of SEQ ID NO: 1;    (d) a polypeptide comprising amino acids 100 to 110 of SEQ ID NO: 1;    (e) a polypeptide comprising amino acids 114 to 130 of SEQ ID NO: 1;    (f) a polypeptide comprising at least one conservative amino acid substitution in the sequence of polypeptides (a-e); and    (g) the amides thereof.    
     
     
         16 . A vector comprising a polynucleotide of  claim 1  operably linked to control sequences which direct the expression of the polynucleotide.  
     
     
         17 . A vector comprising a polyfnucleotide of  claim 2  operably linked to control sequences which direct the expression of the polynucleotide.  
     
     
         18 . A vector comprising a polynucleotide of  claim 3  operably linked to control sequences which direct the expression of the polynucleotide.  
     
     
         19 . A vector comprising a polynucleotide of  claim 4  operably linked to control sequences which direct the expression of the polynucleotide.  
     
     
         20 . A vector comprising a polynucleotide of  claim 5  operably linked to control sequences which direct the expression of the polynucleotide.  
     
     
         21 . A vector comprising a polynucleotide of  claim 6  operably linked to control sequences which direct the expression of the polynucleotide.  
     
     
         22 . A vector comprising a polynucleotide of  claim 7  operably linked to control sequences which direct the expression of the polynucleotide.  
     
     
         23 . A host cell transformed with a vector of  claim 16 .  
     
     
         24 . A host cell transformed with a vector of  claim 17 .  
     
     
         25 . A host cell transformed with a vector of  claim 18 .  
     
     
         26 . A host cell transformed with a vector of  claim 19 .  
     
     
         27 . A host cell transformed with a vector of  claim 20 .  
     
     
         28 . A host cell transformed with a vector of  claim 21 .  
     
     
         29 . A host cell transformed with a vector of  claim 22 .  
     
     
         30 . A pharmaceutical composition comprising a polypeptide of  claim 8  and a pharmaceutically acceptable carrier.  
     
     
         31 . A pharmaceutical composition comprising a polypeptide of  claim 9  and a pharmaceutically acceptable carrier.  
     
     
         32 . A pharmaceutical composition comprising a polypeptide of  claim 10  and a pharmaceutically acceptable carrier.  
     
     
         33 . A pharmaceutical composition comprising a polypeptide of  claim 11  and a pharmaceutically acceptable carrier.  
     
     
         34 . A pharmaceutical composition comprising a polypeptide of  claim 14  and a pharmaceutically acceptable carrier.  
     
     
         35 . A pharmaceutical composition comprising a polypeptide of  claim 15  and a pharmaceutically acceptable carrier.  
     
     
         36 . A method of treating a neurological disease or homeostatic dysfunction or controlling the production of a homeostatic regulatory hormone comprising introducing an effective amount of the composition of  claim 18  into a mammal in need of such treatment.  
     
     
         37 . An antibody that immunoreacts with an isolated mammalian H35 protein.  
     
     
         38 . An antibody of  claim 37  which is a monoclonal antibody.  
     
     
         39 . A kit for detecting the presence of an H35 protein in a mammalian sample comprising an antibody which immunoreacts with a mammalian H35 protein or with a polypeptide of  claim 10  in an amount sufficient for at least one assay and suitable packaging material.  
     
     
         40 . A kit for detecting the presence of an H35 protein in a mammalian sample comprising an antibody which immunoreacts with a mammalian H35 protein or with a polypeptide of  claim 11  in an amount sufficient for at least one assay and suitable packaging material.  
     
     
         41 . A kit for detecting the presence of an H35 protein in a mammalian sample comprising an antibody which immunoreacts with a mammalian H35 protein or with a polypeptide of  claim 14  in an amount sufficient for at least one assay and suitable packaging material.  
     
     
         42 . A kit for detecting the presence of an H35 protein in a mammalian sample comprising an antibody which immunoreacts with a mammalian H35 protein or with a polypeptide of  claim 15  in an amount sufficient for at least one assay and suitable packaging material.  
     
     
         43 . The kit of  claim 39  further comprising a detecting antibody which binds to the anti-H35 antibody.  
     
     
         44 . The kit of  claim 43  wherein the detecting antibody is labeled.  
     
     
         45 . The kit of  claim 44  wherein the label comprises enzymes, radioisotopes, fluorescent compounds, colloidal metals, chemiluminescent compounds, phosphorescent compounds, or bioluminescent compounds.  
     
     
         46 . A kit for detecting the presence of genes encoding an H35 protein comprising a polynucleotide of  claim 1 , or fragment thereof having at least 10 contiguous bases, in an amount sufficient for at least one assay, and suitable packaging material.  
     
     
         47 . A method for detecting the presence of a nucleic acid encoding an H35 protein in a mammalian sample, comprising the steps of: 
 (a) hybridizing a polynucleotide of  claim 1 , or fragment thereof having at least 10 contiguous bases, with the nucleic acid of the sample; and    (b) detecting the presence of the hybridization product.    
     
     
         48 . A method of detecting an H35 antigen in a mammalian sample comprising the steps of: 
 (a) contacting the sample with an anti-H35 antibody which immunoreacts with a hypocretin polypeptide; and    (b) detecting the presence of an immunoreaction complex.    
     
     
         49 . The method of  claim 48  wherein said complex is detected by admixing said immunoreaction complex with a detecting antibody capable of immunoreacting with said anti-H35 antibody.  
     
     
         50 . The method of  claim 49  wherein the detecting antibody is labeled.  
     
     
         51 . The method of  claim 48  wherein the anti-H35 antibody is immobilized on a solid support.  
     
     
         52 . The method of  claim 48  wherein the sample comprises cells.  
     
     
         53 . The method of  claim 52  wherein the cells are peripheral blood mononuclear cells.  
     
     
         54 . The method of  claim 48  wherein the immunoreaction complex of step (b) is detected by flow cytometry.  
     
     
         55 . The method of  claim 48  wherein the immunoreaction complex of step (b) is detected by ELISA.  
     
     
         56 . A method of  claim 48  wherein the immunoreaction complex of step (b) is detected by immunoblot analysis.  
     
     
         57 . A polynucleotide comprising the sequence of SEQ ID NO: 5.  
     
     
         58 . A vector comprising the sequence of SEQ ID NO: 5.  
     
     
         59 . A host cell transfected with the vector of  claim 58.