Hybrid proteins that migrate retrogradely and transynaptically into the CNS

Abstract

The non-toxic proteolytic C fragment of tetanus toxin (TTC peptide) has the same ability to bind nerve cells and be retrogradely transported through a synapse as the native toxin. A hybrid protein encoded by the IacZ-TTC gene fusion retains the biological functions of both proteins in vivo, i.e. retrograde transynaptic transport of the TTC fragment and β-gal enzymatic activity. After intramuscular injection, enzymatic activity could be detected in motoneurons and connected neurons of the brainstem areas. This strategy is useful for the delivery of a biological activity to neurons from the periphery to the central nervous system. Such a hybrid protein can also be used to map synaptic connections between neural cells.

1 - 11 . (canceled) 
     
     
         12 . A method for in vivo delivery of a desired composition into human or animal central nervous system (CNS) or spinal cord, wherein the method comprises administering to the human or animal a composition comprising a vector containing nucleotides encoding, a non-toxic, proteolytic fragment of tetanus toxin (TT) in association with at least a molecule having a biological function and said composition is capable of in vivo retrograde axonal transport and transynaptic transport into the CNS or the spinal cord of the human or animal and of being delivered at different areas of the CNS or the spinal cord, wherein the vector is capable of in vivo expression in a muscle. 
     
     
         13 . The method according to  claim 12 , wherein said vector comprises a promoter and an enhancer capable of expressing the nucleotides contained in said vector in the muscle. 
     
     
         14 . The method according to  claim 13 , wherein said vector is the plasmid pCMV-LacZ-TTC which has been deposited at the C.N.C.M. on Aug. 12, 1997, under the registration number I-11912. 
     
     
         15 . The method according to  claim 12 , wherein said vector is administered into the muscle. 
     
     
         16 . The method according to  claim 12 , wherein the molecule is a nucleotide encoding for a protein or a polypeptide linked chemically to the fragment of tetanus toxin and being transported and expressed directly in neurons. 
     
     
         17 - 26 . (canceled) 
     
     
         27 . A method for in vivo delivery of a desired composition into human or animal central nervous system (CNS) or spinal cord, wherein the method comprises administering to the human or animal a composition comprising a cell or a vector containing nucleotides encoding a non-toxic, proteolytic fragment of tetanus toxin (TT) in association with at least a molecule having a biological function and said composition is capable of in vivo retrograde axonal transport and transynaptic transport into the CNS or the spinal cord of the human or animal and of being delivered at different areas of the CNS or the spinal cord, wherein the cell or vector is capable of in vivo expression in neuronal cells or precursor of neuronal cells and wherein said cell is reimplanted into the CNS or spinal cord. 
     
     
         28 . The method according to  claim 27  wherein said cell or vector comprises a promoter and an enhancer capable of expressing the nucleotides contained in said cell in neuronal cells or precursors of neuronal cells. 
     
     
         29 . The method according to  claim 27 , wherein the molecule is a nucleotide encoding for a protein or a polypeptide linked chemically to the fragment of tetanus toxin and being expressed directly in neurons. 
     
     
         30 . The method according to  claim 27 , wherein the molecule is a nucleotide encoding for a protein or a polypeptide linked chemically to the fragment of tetanus toxin and being expressed directly in neurons. 
     
     
         31 . (canceled)