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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

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An Erratum to this article was published on 21 March 2015

Abstract

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 of CS-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 inactive TeTx 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 neurons to 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.

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Acknowledgments

This study was supported by a Research Professorship plus Principal Investigator award to Prof. J. O. Dolly from Science Foundation Ireland and Programme for Research in Third Level Institutions (PRTLI) Cycle 4 from the Higher Education Authority of Ireland for the Neuroscience section of Target-Driven Therapeutics and Theranostics (Dr. Saak Ovsepian and Prof. J. Oliver Dolly).

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    Authors and Affiliations

    1. International Centre for Neurotherapeutics, Dublin City University, Dublin 9, Ireland

      Saak V. Ovespian, MacDara Bodeker, Valerie B. O’Leary, Gary W. Lawrence & J. Oliver Dolly

    2. German Centre for Neurodegenerative Disease (DZNE), Ludwig-Maximilian-Universität München, Zentrum für Neuropathologie, 81377, Munich, Germany

      Saak V. Ovespian

    3. Life Sciences Department, Institute of Technology, Sligo, Ireland

      MacDara Bodeker

    4. Institute of Radiation Biology, Helmholtz Zentrum München, 85764, Neuherberg, Germany

      Valerie B. O’Leary

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    1. Saak V. Ovespian
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    Correspondence to Saak V. Ovespian.

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    S. V. Ovespian and M. Bodeker equally contributed to this work.

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    Ovespian, S.V., Bodeker, M., O’Leary, V.B. et al. 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. Brain Struct Funct 220, 1825–1838 (2015). https://doi.org/10.1007/s00429-015-1004-0

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