caa a22 4500 60620234X CHVBK 20210128100952.0 cr unu---uuuuu 210128e20150501xx s 000 0 eng 10.1007/s00429-015-1004-0 doi (NATIONALLICENCE)springer-10.1007/s00429-015-1004-0 Internalization and retrograde axonal trafficking of tetanus toxin in motor neurons and trans-synaptic propagation at central synapses exceed those of its C-terminal-binding fragments [Elektronische Daten] [Saak Ovespian, MacDara Bodeker, Valerie O'Leary, Gary Lawrence, J. Oliver Dolly] The prominent tropism of tetanus toxin (TeTx) towards peripheral nerves with retrograde transport and transfer to central neurons render it an invaluable probe for exploring fundamental neuronal processes such as endocytosis, retrograde trafficking and trans-synaptic transport to central neurons. While the specificity of TeTx to nerve cells has been attributed to its binding domains (HC and HCC), molecular determinants of the long-range trafficking that ensure its central delivery and induction of spastic paralysis remain elusive. Here, we report that a protease-inactive TeTx mutant (TeTIM) fused to core streptavidin (CS) proved superior to CS-HC and CS-HCC fragments in antagonizing the internalization of the active toxin in cultured spinal cord neurons. Also, in comparison to CS-HC and CS-HCC, CS-TeTIM undergoes faster clearance from motor nerve terminals after peripheral injection, and is detected in a greater number of neurons in the spinal cord and brain stem ipsi-lateral to the administration site. Consistent with trans-synaptic transfer from motor neurons to inter-neurons, CS-TeTIM infiltrated non-cholinergic cells in the spinal cord; in contrast, the retrograde spread ofCS-HC was largely restricted to neurons stained for choline acetyltransferase. Peripheral injection of CS-TeTIM conjugated to a lentivirus encoding mutated SNAP-25, resistant to cleavage by botulinum neurotoxin A, E and C1, rendered spontaneous excitatory postsynaptic currents in motor neurons resilient to challenge by type A toxin in vitro, whereas the same virus conjugated to CS-HC proved ineffective. These findings indicate that full-length inactiveTeTx greatly exceeds HC and HCC in targeting and invading motor nerve terminals at the periphery and exploits more efficiently the retrograde transport and trans-synaptic transfer mechanisms of motor neuronsto arrive at central neurons. Such qualities render TeTIM a more suitable research probe and neuron-targeting vehicle for retro-axonal delivery of viral vectors to the CNS. Springer-Verlag Berlin Heidelberg, 2015 Retrograde trafficking nationallicence Motor neuron nationallicence Trans-synaptic transport nationallicence SNARE protein nationallicence Neuronal targeting nationallicence Tetanus toxin nationallicence Ovespian Saak International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut Bodeker MacDara International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut O'Leary Valerie International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut Lawrence Gary International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut Oliver Dolly J. International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut Brain Structure and Function Springer Berlin Heidelberg 220/3(2015-05-01), 1825-1838 1863-2653 220:3<1825 2015 220 429 https://doi.org/10.1007/s00429-015-1004-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00429-015-1004-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ovespian Saak International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut NATIONALLICENCE 700 1- Bodeker MacDara International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut NATIONALLICENCE 700 1- O'Leary Valerie International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut NATIONALLICENCE 700 1- Lawrence Gary International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut NATIONALLICENCE 700 1- Oliver Dolly J. International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland aut NATIONALLICENCE 773 0- Brain Structure and Function Springer Berlin Heidelberg 220/3(2015-05-01), 1825-1838 1863-2653 220:3<1825 2015 220 429