gms | German Medical Science

64th Annual Meeting of the German Society of Neurosurgery (DGNC)

German Society of Neurosurgery (DGNC)

26 - 29 May 2013, Düsseldorf

Clostridium botulinum peptide – C3bot – promotes outgrowth of motor cortical fibres into spinal cord ex vivo

Meeting Abstract

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  • Jana Glumm - Institut für Zell- und Neurobiologie, Fächerverbund für Anatomie, Charité-Universitätsmedizin Berlin; Klinik für Neurochirurgie, Helios Klinikum Berlin-Buch, Deutschland
  • Martin Pohland - Institut für Zell- und Neurobiologie, Fächerverbund für Anatomie, Charité-Universitätsmedizin Berlin
  • Jürgen Kiwit - Klinik für Neurochirurgie, Helios Klinikum Berlin-Buch, Deutschland

Deutsche Gesellschaft für Neurochirurgie. 64. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC). Düsseldorf, 26.-29.05.2013. Düsseldorf: German Medical Science GMS Publishing House; 2013. DocP 069

doi: 10.3205/13dgnc486, urn:nbn:de:0183-13dgnc4865

Published: May 21, 2013

© 2013 Glumm et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.



Objective: A variety of SCI models exists in vivo, but due to their complexity, comparability and clinical practicability is extremely difficult. During the last years we have established a new coculture method combining murine motor cortical (MC) and spinal cord (SC) slices. We have further investigated motor cortical regeneration and sprouting to the spinal cord and characterized the ingrowing neurons and invading cells. We present here our data on enhanced outgrowth by treatment with C3bot, known for enhancing recovery of descending spinal cord fibers after dorsal hemisection or compression SC injury in mice.

Method: MC prepared from postnatal Bl/6.GFP P0-P3 pups was cut along the coronal axis. SC was dissected from postnatal C57Bl/6 P0-P3 pups and subsequently chopped along a sagittal longitudinal plane. Thereafter, the medial MC zone was oriented to the rostral end of the SC and incubated up to 28 days. To analyse the effects of potential outgrowth compounds, C3bot – Clostridium botulinum C3-protein-derived 29-amino-acid fragment (154–182) was added to the coculture incubation medium directly after slice preparation and every 2 DIV thereafter at a dilution of 300 nM in PBS. Control slice cultures received PBS or NGF. To analyze SC in growth we did immunhistochemical staining, fluorescence as well as live imaging and electrophysiological analyses by signal transduction of intracellular calcium concentrations.

Results: Using our slice culture setup, we confirmed the ability for motor cortical axonal outgrowth. Treatment with C3bot promoted outgrowth of motor cortical fibres into spinal cord significantly. Pre- and postsynaptic stainings reveal the formation of new synaptic connections and SC reinnervation, confirmed by our electrophysiological analyses with strong excitation of spinal neurons and we calculated an average nerve conduction velocity of 40 m/s.

Conclusions: We describe further our GFP cytoarchitecture-preserving slice coculture technique to analyse regeneration between MC and SC ex vivo. Treatment with Clostridium botulinum peptide enhanced MC sprouting significantly. By the use of pharmacological modulating factors we can enhance cortical outgrowth and boost new synaptic connections in to the spinal cord. Thus, this new slice coculture model provides an important tool for investigating a variety of questions in the field of in vitro neuronal regeneration by examining the influences of different cell types and screening of factors on corticospinal tract regeneration ex vivo.