title: Minimal essential length of Clostridium botulinum C3 peptides to enhance neuronal regenerative growth and connectivity in a non-enzymatic mode.
abstract: C3 ADP-ribosyltransferase is a valuable tool to study Rho-dependent cellular processes. In the current study we investigated the impact of enzyme-deficient peptides derived from Clostridium botulinum C3 transferase in the context of neuronal process elongation and branching, synaptic connectivity, and putative beneficial effects on functional outcome following traumatic injury to the CNS. By screening a range of peptidic fragments, we identified three short peptides from C3bot that promoted axon and dendrite outgrowth in cultivated hippocampal neurons. Furthermore, one of these fragments, a 26-amino acid peptide covering the residues 156-181 enhanced synaptic connectivity in primary hippocampal culture. This peptide was also effective to foster axon outgrowth and re-innervation in organotypical brain slice culture. To evaluate the potential of the 26mer to foster repair mechanisms after CNS injury we applied this peptide to mice subjected to spinal cord injury by either compression impact or hemisection. A single local administration at the site of the lesion improved locomotor recovery. In addition, histological analysis revealed an increased serotonergic input to lumbar motoneurons in treated compared with control mice. Pull-down assays showed that lesion-induced up-regulation of RhoA activity within the spinal cord was largely blocked by C3bot peptides despite the lack of enzymatic activity.
authors.affiliation: Center for Anatomy, Functional Cell Biology, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Loske
authors.forename: Francesco
authors.initials: F
authors.lastname: Boato
authors.forename: Sven
authors.initials: S
authors.lastname: Hendrix
authors.forename: Johannes
authors.initials: J
authors.lastname: Piepgras
authors.forename: Ingo
authors.initials: I
authors.lastname: Just
authors.forename: Gudrun
authors.initials: G
authors.lastname: Ahnert-Hilger
authors.forename: Markus
authors.initials: M
authors.lastname: Höltje
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry* / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D001905:Botulinum Toxins / Q000737:chemistry* / Q000494:pharmacology*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D059330:GABAergic Neurons / Q000187:drug effects / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D008124:Locomotion / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000166:cytology*; D009924:Organ Culture Techniques; D010455:Peptides / Q000494:pharmacology / Q000627:therapeutic use; D059326:Serotonergic Neurons / Q000187:drug effects / Q000502:physiology; D012701:Serotonin / Q000378:metabolism; D013119:Spinal Cord Injuries / Q000188:drug therapy* / Q000209:etiology / Q000473:pathology; D058630:Spinal Cord Regeneration / Q000187:drug effects*; D013569:Synapses / Q000187:drug effects*; D013997:Time Factors; D014162:Transfection; D053675:Versicans / Q000378:metabolism; D050495:Vesicular Inhibitory Amino Acid Transport Proteins / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism
chemical_list: D010455:Peptides; C118623:Slc32a1 protein, rat; C508324:Vcan protein, rat; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; D053675:Versicans; D049452:Green Fluorescent Proteins; D012701:Serotonin; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D001905:Botulinum Toxins; D020741:rho GTP-Binding Proteins