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Botulinum Neurotoxin Light Chains Expressed by Defective Herpes Simplex Virus Type-1 Vectors Cleave SNARE Proteins and Inhibit CGRP Release in Rat Sensory Neurons

AuthorsJoussain, Charles; Le Coz, Olivier; Pichugin, , Andrey; Marconi, Peggy; Lim, Filip ; Sicurella, Mariaconcetta; Salonia, Andrea; Montorsi, Francesco; Wandosell, Francisco G.; Foster, Keith; Giuliano, François; Epstein, Alberto L; Aranda Muñoz, Alejandro
KeywordsHSV-1 amplicon vectors
Transgenic botulinum neurotoxins
Light chains
Sensory neurons
SNARE proteins
Issue Date19-Feb-2019
CitationToxins 11 (2019)
AbstractA set of herpes simplex virus type 1 (HSV-1) amplicon vectors expressing the light chains (LC) of botulinum neurotoxins (BoNT) A, B, C, D, E and F was constructed. Their properties have been assessed in primary cultures of rat embryonic dorsal root ganglia (DRG) neurons, and in organotypic cultures of explanted DRG from adult rats. Following infection of primary cultures of rat embryonic DRG neurons, the different BoNT LC induced efficient cleavage of their corresponding target Soluble N-ethylmaleimide-sensitive-factor Attachment protein Receptor (SNARE) protein (VAMP, SNAP25, syntaxin). A similar effect was observed following infection by BoNT-A LC of organotypic cultures of adult rat DRG. To quantify and compare the functional activities of the different BoNT LC, the inhibition of calcitonin gene-related protein (CGRP) secretion was assessed in DRG neurons following infection by the different vectors. All BoNT-LC were able to inhibit CGRP secretion although to different levels. Vectors expressing BoNT-F LC displayed the highest inhibitory activity, while those expressing BoNT-D and -E LC induced a significantly lower CGRP release inhibition. Cleavage of SNARE proteins and inhibition of CGRP release could be detected in neuron cultures infected at less than one transducing unit (TU) per neuron, showing the extreme efficacy of these vectors. To our knowledge this is the first study investigating the impact of vector-expressed transgenic BoNT LC in sensory neurons.
Publisher version (URL)http://dx.doi.org/10.3390/toxins11020123
Identifiersdoi: 10.3390/toxins11020123
issn: 2072-6651
Appears in Collections:(CBM) Artículos
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