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J Neurochem. 2016 Oct;139(2):234-244. doi: 10.1111/jnc.13739. Epub 2016 Aug 9.

The intermediate filament protein vimentin is essential for axonotrophic effects of Clostridium botulinum C3 exoenzyme.

Author information

1
Institute of Integrative Neuroanatomy, Charité-Universitätsmedizin Berlin, Berlin, Germany.
2
Department of Pharmacology, Medical School, University of Ioannina, Ioannina, Greece.
3
Institute of Toxicology, Hannover Medical School (MHH), Hannover, Germany.
4
Institute of Biochemistry & Neuro Cure Cluster of Excellence, Charité - Universitätsmedizin Berlin, Berlin, Germany.
5
Institute of Integrative Neuroanatomy, Charité-Universitätsmedizin Berlin, Berlin, Germany. markus.hoeltje@charite.de.

Abstract

The type III intermediate filament protein vimentin was recently identified to mediate binding and uptake of Clostridium botulinum C3 exoenzyme (C3bot) in two cell lines. Here, we used primary neuronal cultures from vimentin knockout (Vim-/- ) mice to study the impact of vimentin on axonal growth and internalization of C3bot. In contrast to wild type, vimentin knockout neurons were insensitive to C3bot. Application of extracellular vimentin to Vim-/- neurons completely restored the growth-promoting effects of C3bot. In line with this uptake of C3bot into Vim-/- neurons was strongly decreased resulting in reduced ADP-ribosylation of RhoA and B as detected by an antibody recognizing selectively ADP-ribosylated RhoA/B. Again, uptake of C3bot into Vim-/- neurons was rescued by addition of extracellular vimentin. In addition, in purified embryonic stem cell-derived motor neurons that are devoid of glial cells C3bot elicited axonotrophic effects confining neuronal vimentin as a binding partner. Primary neuronal cultures from vimentin knockout (KO) mice were used to study the impact of vimentin on axonal growth and internalization of C3bot. In contrast to wild type, vimentin knockout neurons were insensitive to the axonotrophic effects of C3bot. Application of extracellular vimentin (recombinant vimentin) to vimentin KO neurons completely restored the growth-promoting effects of C3bot. In line with this uptake of C3bot into vimentin KO neurons was strongly decreased resulting in reduced ADP-ribosylation of RhoA and B as detected by an antibody recognizing selectively ADP-ribosylated RhoA/B.

KEYWORDS:

C3 exoenzyme; axon outgrowth; vimentin

PMID:
27419376
DOI:
10.1111/jnc.13739
[Indexed for MEDLINE]
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